EA029372B1 - Pyrazolopyrimidine compounds - Google Patents

Abstract

The present invention provides a compound represented by the following structural formula:or a pharmaceutically acceptable salt thereof, wherein R, R, Rand Rare disclosed in the claims. Also described are pharmaceutical compositions and methods of use thereof.

Description

The invention relates, at least in particular, to a compound represented by the following structural formula:

or its pharmaceutically acceptable salt,

where K ¹ is a -ΝΗ-ssch-soo;

Su represents a C ₃ -C ₄ -cycloalkyl, optionally having as substituents one

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or two groups selected from alkyl and hydroxyl;

K ² is -O-pyridinyl, -HN- (C ₂ -C ₆ ) hydroxyalkyl, optionally having cyclopropyl or isopropyl as a substituent, or -XH- (C ₃ -C ₆ ) cycloalkyl, optionally having hydroxyl as a substituent or (CrClHydroxylalkyl;

K ^{4 is} selected from a hydrogen atom, a halogen atom, and (C _g C ₃ ) alkyl;

K ^a is cyclopropyl.

In a preferred embodiment of the invention, K ⁴ is a chlorine atom or methyl. According to one embodiment of the present invention, a pharmaceutical composition is provided comprising a pharmaceutically acceptable carrier or diluent and a compound represented by the structural formula (I) described above, or a pharmaceutically acceptable salt thereof.

According to another embodiment of the present invention, a method is proposed for treating a subject suffering from cancer, comprising administering to the subject an effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof.

According to another embodiment of the present invention, a method is proposed for inhibiting TTC activity in a subject in need of inhibiting TTC activity, comprising administering to the subject an effective amount of a compound represented by structural formula (I) or a pharmaceutically acceptable salt thereof.

According to another embodiment of the present invention, the use of a compound represented by the structural formula (I) or a pharmaceutically acceptable salt thereof in therapy is provided. In some embodiments of the invention, said therapy is directed to the treatment of a subject suffering from cancer. Alternatively, this therapy is aimed at inhibiting the activity of TTC in a subject in need of inhibiting the activity of TTC.

According to another embodiment of the present invention, there is provided the use of a compound represented by the structural formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a subject suffering from cancer.

According to another embodiment of the present invention, there is provided the use of a compound represented by the structural formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament intended to inhibit the activity of TTC in a subject in need of inhibiting the activity of TTA.

Detailed Description of the Invention

According to one embodiment, the present invention relates to a compound represented by the structural formula (I), or a pharmaceutically acceptable salt thereof. The invention also relates to compounds described using the structural formula or name in the Examples section, and includes both neutral forms and pharmaceutically acceptable salts of these compounds. The invention also relates to the treatment of these compounds and / or using these compounds (including their neutral forms and pharmaceutically acceptable salts), such as described in this document. The following are specific examples of compounds according to the present invention:

or its pharmaceutically acceptable salt;

or its pharmaceutically acceptable salt;

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or its pharmaceutically acceptable salt;

or its pharmaceutically acceptable salt.

The term "alkyl", used alone or as part of a larger group, such as "hydroxyalkyl" and the like, means a saturated aliphatic monovalent hydrocarbon radical with a normal or branched chain. Unless otherwise indicated, the alkyl group usually contains from 1 to 6 carbon atoms, i.e. represents (C ₁ -C ₆ ) alkyl. In the context of this description, the group "(C ₁ -C ₆ ) alkyl" means a radical with a normal or branched chain, having from 1 to 6 carbon atoms.

The term "cycloalkyl" means a saturated aliphatic cyclic hydrocarbon radical, optionally containing one or more double bonds. This radical can be a monocyclic ring, a bicyclic ring (for example, a bicyclic bridge ring), a polycyclic (for example, a tricyclic ring) or a condensed ring. For example, monocyclic (C ₃ -C ₇ ) cycloalkyl means a radical having from 3 to 7 carbon atoms forming a monocyclic ring; (C ₃ -C ₇ ) cycloalkyl includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

Some compounds described herein may exist in various stereoisomeric or tautomeric forms. Stereoisomers are compounds that differ only in spatial arrangement. When a compound according to the invention is described using a name or structure without indicating stereochemistry, it is understood that the name or structure includes all possible stereoisomers, tautomers, geometric isomers, or combinations thereof.

When the geometric isomer is described using a name or structure, it is understood that the isomeric purity of the named or illustrated geometric isomer is at least 60, 70, 80, 90, 99, or 99.9% by weight. The isomeric purity of the geometric isomer is determined by dividing the mass of the named or depicted geometric isomer present in the mixture by the total mass of all the geometric isomers present in the mixture.

A racemic mixture is a mixture containing 50% of one enantiomer and 50% of the other enantiomer. The present invention includes all enantiomerically pure, enantiomerically enriched, diastereomerically pure, diastereomerically enriched and racemic mixtures and diastereoisomeric mixtures of the compounds described in this patent.

Enantiomeric and diastereomeric mixtures can be separated into their individual enantiomers or stereoisomers using known techniques such as gas chromatography on a chiral phase, high performance liquid chromatography on a chiral phase, crystallization of the compound as a complex with a chiral salt or crystallization in a chiral solvent. Enantiomers and diastereoisomers can also be obtained from diastereomerically or enantiomerically pure intermediates, reagents and catalysts using known asymmetric synthesis techniques.

When a compound is described using a name or structure that corresponds to an individual enantiomer, the optical purity of the compound, unless otherwise indicated, is at least 60, 70, 80, 90, 99, or 99.9% (in this case, the compound is also called " enantiomerically pure "). Optical purity is determined by dividing the mass of the named or depicted enantiomer present in the mixture by the total mass of both enantiomers present in the mixture.

When the stereochemistry of a compound according to the invention is indicated in the name or structure and this name or structure includes more than one stereoisomer (for example, a diastereoisomeric pair), it is implied that one of these stereoisomers or any mixture of these stereoisomers is included.

ditch It is also understood that the stereoisomeric purity of the stereoisomers mentioned or shown is at least 60, 70, 80, 90, 99 or 99.9% by weight. Stereoisomeric purity in this case is determined by dividing the total mass of stereoisomers present in the mixture, which correspond to a given name or structure, by the total mass of all stereoisomers present in the mixture.

The present invention provides pharmaceutically acceptable salts of the compounds of the invention. The compounds of the present invention contain basic amino groups and therefore can form pharmaceutically acceptable salts with pharmaceutically acceptable acid (s). Suitable pharmaceutically acceptable acid addition salts of the compounds described in this invention include salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, nitric and sulfuric acids) and organic acids (such as acetic acid, benzenesulfonic, benzoic, methanesulfonic and -toluenesulfonic acid). Compounds of the present invention containing acidic groups, such as carboxyl, can form pharmaceutically acceptable salts with pharmaceutically acceptable (s) base (s).

Suitable pharmaceutically acceptable base salts include ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts). Compounds containing a quaternary ammonium group also contain a counter-anion such as chloride, bromide, iodide, acetate, perchlorate, and the like. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates and salts with amino acids such as glutamic acid.

The compounds described in this patent can inhibit various kinases, including TTC. Accordingly, in most cases, the compounds described herein can be used to treat diseases or conditions associated with such kinases. According to some embodiments of the present invention, the compounds described herein can inhibit the protein kinase TTC.

In one embodiment of the present invention, the compounds described herein are inhibitors of the protein kinase TTC and can be used to treat diseases such as cancer associated with such (their) kinase (s).

Another aspect of the present invention relates to a method of treating a subject suffering from cancer, comprising administering to the subject an effective amount of a compound described herein. According to one embodiment of the present invention, the compounds described herein inhibit tumor growth. For example, the compounds described in this document inhibit the growth of a tumor that overexpresses TTC.

Types of cancer that can be treated (including reducing the likelihood of relapse) by the methods of the present invention include lung cancer, breast cancer, colon cancer, brain cancer, neuroblastoma, prostate cancer, melanoma, glioblastoma multiforme, ovarian cancer, lymphoma, leukemia , melanoma, sarcoma, paraneoplasia, osteosarcoma, germina, glioma and mesothelioma. In one of the embodiments of the present invention, the cancer is selected from leukemia, acute myeloid leukemia, chronic myelogenous leukemia, breast cancer, brain cancer, colon cancer, colorectal cancer, head and neck cancer, hepatocellular cancer, lung adenocarcinoma, metastatic melanoma, pancreatic cancer, prostate cancer, ovarian cancer and kidney cancer. In one embodiment of the present invention, the cancer is lung cancer, colon cancer, brain cancer, neuroblastoma, prostate cancer, melanoma, glioblastoma multiforme, or ovarian cancer. According to another embodiment of the present invention, the cancer is pancreatic cancer, prostate cancer, lung cancer, melanoma, breast cancer, colon cancer, or ovarian cancer. According to another embodiment of the present invention, the cancer is breast cancer, colon cancer and ovarian cancer. According to another embodiment of the present invention, the cancer is breast cancer. According to another embodiment of the present invention, the cancer is a basal subtype of breast cancer or a lumenal B subtype of breast cancer. In yet another embodiment of the present invention, the cancer is a basal subtype of breast cancer that overexpresses TTC. According to another embodiment of the present invention, the basal subtype of breast cancer is EK (estrogen receptor), HEK2 and RK (progesterone receptor) negative breast cancer. According to another embodiment of the present invention, the cancer is soft tissue cancer. "Soft tissue cancer" is a term used in the art that includes tumors derived from any soft tissue of the body. Such soft tissue connects, supports or surrounds various structures and organs of the body and includes, without limitation, smooth muscle, skeletal muscle, tendons, fibrous tissue, adipose tissue, blood and lymphatic vessels, perivascular tissue, nerves, mesenchymal cells and synovial tissue. Accordingly, soft tissue cancer may be a cancer of adipose tissue, muscle tissue, nervous tissue, articular tissue, blood vessels, lymphatic vessels, and fibrous tissues. Soft tissue cancer can be dobrokak- 4 029372

or malignant. Usually, malignant soft tissue cancer is called sarcoma or soft tissue sarcoma. There are many types of soft tissue, including lipoma, lipoblastoma, hibernation, liposarcoma tumor, hemangioperiticoma, hemangioendothelioma, angiosarcoma, Kaposi's sarcoma, lymphangioma, fibroma, elastofibroma, superficial fibromatosis, fibrous histiocytoma, fibrosarcoma, fibromatosis, protrude th dermatofibrosarkomu (ΌΡδΡ), malignant fibrous histiocytoma (MPH) mixes zernistokletochnuyu tumor, malignant mesenchymal alveolar soft tissue sarcoma, epithelioid sarcoma, clear cell sarcoma and desmoplastic small cell tumor. In a specific embodiment of the present invention, soft tissue cancer is a sarcoma selected from the group consisting of fibrosarcoma, gastrointestinal sarcoma, leiomyosarcoma, dedifferentiated liposarcoma, pleomorphic liposarcoma, malignant fibrous histiocytoma, circular cell saroma.

According to some embodiments, the present invention provides methods for inhibiting the growth of cells initiating tumor development or reducing the likelihood of cancer recurring in a subject who is receiving anti-cancer therapy. This method includes the steps of:

a) assessing the condition of the subject to determine whether the cancer is in remission or not; and

b) if the cancer is in remission, administering to the subject an effective amount of a protein kinase inhibitor TTC (for example, a compound represented by structural formula (I)).

If the cancer is not in remission, the method optionally includes the additional step of continuing anticancer therapy until the cancer goes into remission, and then performing step (b) of administering an effective amount of a protein kinase inhibitor TTC (for example, the compound represented by structural formula (I)) .

In the context of this description, the term "cells initiating the development of a tumor" or "T1C" refers to cells that are present inside certain tumors and possess the ability for self-maintenance and proliferation. These cells are sometimes called cancer stem cells (C8C); it can be observed that C8C has some features characteristic of normal stem cells, including the similarity of the phenotype and function of cancer and normal stem cells. According to some embodiments of the present invention, T1C are characterized in that they are capable of forming tumors after xenotransplantation in immunodeficient mice.

According to some embodiments, the present invention provides methods for inhibiting the growth of cells initiating tumor development, or reducing the likelihood of cancer recurring in a subject having cancer in remission, including administering to the subject an effective amount of a protein kinase inhibitor TTC (eg, a compound represented by the structural formula (I )).

In some embodiments, implementation of the present invention, for example, in cases where the subject receives treatment to reduce the likelihood of cancer recurrence, the subject has already been treated with anticancer therapy. Alternatively, the subject has already been treated with anticancer therapy, and the subject is in remission.

According to some embodiments, the present invention provides methods for treating a subject suffering from cancer, comprising administering to the subject an effective amount of a compound, a compound represented by structural formula (I), in combination with an effective anti-cancer therapy. In one of the embodiments of the invention, the cancer is a metastatic cancer. "Metastatic cancer" is a cancer that spreads from the primary site of its occurrence to other parts of the body.

In another embodiment, the present invention provides a method for treating a subject with a drug resistant cancer. "Drug-resistant cancer" is a cancer that is not susceptible to one, two, three, four, five or more drugs that are commonly used to treat cancer. In one of the embodiments of the present invention, drug resistant cancer is caused by the growth of cells initiating the development of a tumor.

To assess the condition of the subject, which allows to determine whether the cancer is in remission or not, suitable techniques known in the art can be used. For example, tumor size and / or tumor markers, usually proteins associated with tumors, can be monitored to determine the condition of the cancer. Tumor size can be controlled using imaging techniques such as x-rays, MRI (magnetic resonance imaging), CAT 8eai (computerized axial tomography), ultrasound, mammography, PET (positron emission tomography) and the like, or by biopsy.

In the described methods, for example in methods including co-administration, the anticancer therapy is selected from the group consisting of surgery, radiation therapy, immunotherapy, endocrine therapy, gene therapy and the administration of an anticancer agent. Alternatively, anticancer therapy is radiation therapy. Alternatively, anticancer therapy

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is an immunotherapy. Alternatively, anticancer therapy is the administration of an anticancer agent. Alternatively, anticancer therapy is a surgical procedure.

Radiation therapy is the use of radiation to kill cancer cells, destroy a cancer tumor or treat cancer. Typical radiation therapy includes, without limitation, gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy and radioisotope therapy (i.e., systemic radioisotope therapy).

Endocrine therapy is a treatment that adds, blocks or removes hormones. For example, chemotherapeutic agents that can block estrogen production or activity are used to treat breast cancer. In addition, hormonal stimulation of the immune system is used to treat specific types of cancer, such as renal cell carcinoma and melanoma. In one embodiment of the present invention, endocrine therapy involves the administration of natural hormones, synthetic hormones or other synthetic molecules that can block or enhance the production of the body's natural hormones. In another embodiment of the present invention, endocrine therapy involves removing a gland that produces a specific hormone.

In the context of this description, gene therapy is the introduction of genes into the cells and biological tissues of a subject in order to treat diseases such as cancer. Typical gene therapy includes, without limitation, gene therapy using germ-line cells and somatic gene therapy.

Immunotherapy (also called biologic response modifier therapy, biologic therapy, biotherapy, immune therapy, or biologic therapy) is a treatment that uses immune system components to combat the disease. Immunotherapy can help the immune system recognize cancer cells or increase the response against cancer cells. Immunotherapy includes active and passive immunotherapy. Active immunotherapy stimulates the body’s own immune system, whereas passive immunotherapy usually uses components of the immune system that are obtained outside the body.

Examples of active immunotherapy include, without limitation, vaccines, including cancer vaccines, vaccines against tumor cells (autologous or allogeneic), vaccines based on dendritic cells, antigenic vaccines, anti-idiotypic vaccines, DNA vaccines, viral vaccines or lymphocyte-based vaccines, infiltrating a tumor (T1b), in combination with interleukin 2 (1Ь-2) or cell therapy with lymphokine-activated killer cells (LAK).

Examples of passive immunotherapy include, without limitation, monoclonal antibodies and targeted therapy using toxins. Monoclonal antibodies include naked antibodies and conjugated monoclonal antibodies (also called labeled (! Addeb, LaFell) or loaded antibodies). Bare monoclonal antibodies do not have a drug or radioactive substance attached to them, while conjugated monoclonal antibodies are connected, for example, with a chemotherapeutic drug (antibodies labeled with a chemotherapeutic agent (sietoleber), a radioactive particle (radiolabeled antibodies) or a toxin (immunotoxins). Examples of naked monoclonal antibodies used as drugs include, without limitation, rituximab (Rituxan), an antibody against the antigen SE2 (). used to treat, for example, B-cell non-Hodgkin's lymphoma; Trastuzumab (Herceptin), an antibody against the HEK2 protein, used to treat, for example, advanced breast cancer; alemtuzumab (Kampas), an antibody against CE25 antigen, used to treat, for example, B-cell chronic lymphocytic leukemia (ALL); cetuximab (Erbitux), an antibody against ESCA (epidermal growth factor receptor) protein, used, for example, in combination with irinotecan to treat, for example, advanced colorectal cancer and malignant neoplasms of the head and neck; and bevacizumab (Avastin) for anti-angiogenic therapy directed against the WESR protein, used, for example, in combination with chemotherapy to treat, for example, metastatic colorectal cancer. Examples of conjugated monoclonal antibodies include, without limitation, radiolabeled antibody ibritumomab tiuxetan (zevalin), which delivers radioactivity directly to cancer B-lymphocytes and is used to treat, for example, B-cell non-Hodgkin's lymphoma; radiolabeled antibody tositumomab (Vechhag), which is used to treat, for example, certain types of non-Hodgkin's lymphoma; and gemtuzumab ozogamicin immunotoxin (Mu160), which contains calicheamicin (SaPsNeapist) and is used to treat, for example, acute myelogenous leukemia (AMB); Bb22, conjugated monoclonal antibody for the treatment of, for example, hairy cell leukemia; immunotoxins to treat, for example, leukemias, lymphomas and brain tumors, and radiolabelled antibodies, such as Oiscomasch, for example, for the treatment of colorectal and ovarian cancer, and Pro-313, for example, for the treatment of prostate cancer.

Additional examples of therapeutic antibodies that can be used include

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Herceptin® (trastuzumab) (SsssssssN.SA), which is a humanized monoclonal antibody against HEK2 for the treatment of patients with metastatic breast cancer; ReoPro® (abtsiksimab) (Sssosog). which binds to platelet-type glycoprotein receptor type 1Gb / 111a and prevents thrombosis; Zenapax® (Daclizumab) (Rosier Reagentaseyasak, Switzerland), which is an immunosuppressive humanized monoclonal antibody against CO25 to prevent acute rejection of the renal allograft; Panorex ™ (ΡΑΝΌΚΕΧ ™), which is a murine GcC2a antibody against the 17-HA cell surface antigen (C1aho \ U11ote / Sep1osog); BEC2, which is a murine anti-idiotypic (ΟΌ3 epitope) GDS antibody (GTCoop DrugTech); HMS-S225, which is a chimeric GdS antibody against ESCA (Gtc1ope Sopt); Vitaksin ™ (νΓΤΑΧΓΝ ™), which is a humanized anti-integrin ανβ3 antibody (Arrbeb Motioner Euoloiop / Mebhtshipe); Campas 1Η / ΕΌΡ-03, which is a humanized GcCl antibody against SE52 (Leikokye); §Shay M195, which is a humanized GdS antibody against SE33 (Promellus Ceulchus); Rituxan ™, which is a chimeric GcCl antibody against SE20 (YuES Ryagt / SepeShesk RosNe / Yayuyaki); UMRNOSGOE ™, which is a humanized GdS antibody against SE22 (GtsiposheShsk); AMRNOSYU ™ ™ 90-90 (GttipoteShsk); Lrcoxap (labeled with technetium-99t (Tc-99t); for radionuclide imaging; GtypoteSchsk); No.1 \ aop (v. SE3; Proctor Oe51dp Lak); CM3, which is a humanized anti-GAAM3 antibody (type 3 intercellular adhesion molecule) (GSO3 Phags); UES-114, which is a primatized antibody against SE80 (UES Ryagt / MCTN); Zevalin ™, which is a radioactively isotope-labeled mouse antibody against Ό'.Ό20 (GOES / 3СГгт§ АС); US-131, which is a humanized anti-CO40 antibody (CHOES / EEi); 1EEC-151, which is a primatized antibody against SE4 (EU); YuES-152, which is a primatized antibody against SE23 (GOES / 8e1kadaki); 8MAKT upCO3, which is a humanized GDS against SE3 (Promeche Silent La); 5C1.1, which is a humanized antibody against complement factor C5 (A1xup R); E2E7, which is a humanized antibody against α-α (tumor necrosis factor alpha) (CAT / BA8P); CEP870, which is a humanized PAB fragment against α-α (CeB1B); GOES-151, which is a primatized GcCl antibody against SE4 (GOES Ryagt / 3yyk1e Veesbat); MPH-SE4, which is a human GdS antibody against SE4 (Mebageh / EcAsept); CO20-streptavidin (+ biotin-yttrium 90; No. Xo); CEP571, which is a humanized gcc4 antibody against α-α (CeBeB); LPR-02, which is a humanized antibody against α4β7 (LeikoZk / Sepayes); OyoClope OCT4A, which is a humanized GdS antibody against SE4 (OioBoles); ΑNΤОVΑ ™, which is a humanized GDS anti-CO40 antibody (Wüdep); AETESRE # ^M , which is a humanized GDS anti-URAM (very late antigen-4) antibody (E1ap); and CAT-152, which is a human anti-A-β2 antibody (transforming growth factor β2) (SatBrAllAllS), but not limited to them.

Immunotherapy, which can be used according to the present invention, includes adjuvant immunotherapy. Examples include cytokines, such as granulocyte macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), macrophage inflammatory protein (MHR) -1-alpha, interleukins (including GE-1, Gy-2, Gy-4, ГЬ-6, ГЬ-7, ГЬ-12, ГЕ-15, ГЕ-18, ГЬ-21 and ГЕ-27), tumor necrosis factors (including-α) and interferons (including GH ^ alpha, [ΡΝ-beta and grie-gamma); aluminum hydroxide (a1it); Bacillus Calmetheren (BCG); hemocyanin snails Keubole Be! (Khn); Freund's incomplete adjuvant (GRA); 03-21; ^ ЕΤОX; levamisole and dinitrophenyl (ENR), and combinations thereof, such as, for example, combinations of interleukins, for example, combinations of Gb-2 with other cytokines, such as GΡN-alpha.

Alternatively, the anticancer therapy described herein includes the administration of an anticancer agent. An "anti-cancer agent" is a compound, the introduction of which, in an effective amount, to a subject suffering from cancer, allows to achieve, in part or in essence, one or more of the following results: cessation of development, reduction in the incidence of cancer (for example, reduction of tumor size), inhibiting the rate of cancer and reducing the intensity or improvement of a clinical symptom or indicator associated with cancer (such as tissue or serum components), or an increase in Subject life duration.

An anti-cancer agent suitable for use in the methods described in this invention includes any anti-cancer agents that are approved for the treatment of cancer. In one embodiment of the present invention, the anti-cancer agent includes, without limitation, an anti-target antibody, an angiogenesis inhibitor, an alkylating agent, an antimetabolite, a vinca alkaloid, a taxane, podophyllotoxin, a topoisomerase inhibitor, a hormonal antitumor agent and

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other antitumor agents.

Examples of alkylating agents that can be used in the methods of the present invention include nitrogen mustards (eg, mechlorethamine, cyclophosphamide, chlorambucil, melphalan, etc.), ethyleneimine and methylmelamines (eg, hexamethylmelamine, thiotepa), alkyl sulfonates (eg busulfan), nitrosoureas (for example, carmustine, lomustine, semustine, streptozocin, etc.) or triazenes (decarbazine, etc.), but not limited to them. Examples of antimetabolites that can be used in the methods of the present invention include a folic acid analog (eg, methotrexate) or pyrimidine analogs (eg, fluorouracil, floxuridine, cytarabine), purine analogs (eg, mercaptopurine, thioguanine, pentostatin), but not limited to by them. Examples of plant alkaloids and terpenoids or their derivatives include vinca alkaloids (for example, vincristine, vinblastine, vinorelbine, vindesine), podophyllotoxin and taxanes (for example, paclitaxel, docetaxel), but are not limited to them. Examples of a topoisomerase inhibitor include, but are not limited to, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, and teniposide. Examples of antitumor agents include actinomycin, anthracyclines (eg, doxorubicin, daunorubicin, valrubicin, idarubicin, epirubicin), bleomycin, plicamycin, and mitomycin, but are not limited to them.

According to one embodiment of the invention, anticancer agents that can be used according to the present invention include adriamycin, dactinomycin, bleomycin, vinblastine, cisplatin, acivicin; aclarubicin; acodazole hydrochloride; acronin; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutetimid; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacytidine; Azetepa (a / emera); nitrogenomycin; batimastat; benzodepu; bicalutamide; bisanthrene hydrochloride; bisnafida dimesylate; bizelesin; bleomycin sulfate; sodium brequinar; bopirimine; busulfan; cactinomycin; calusterone; karatsimid; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cededingol; chlorambucil; Cyrolemycin; cladribine; Chrisnatol Mesylate; cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin; desaguanine; desaguanine mesylate; diazikvon (Sha / kcüps); doxorubicin; doxorubicin hydrochloride; droloxifene; Droloxifen citrate; dromostanolone (Dototo81popope) propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrutsin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozol; esorubicin hydrochloride; estramustine; estramustine sodium phosphate; ethanidazole; etoposide; etoposide phosphate; etoprin (e1org1pe); fadrozole hydrochloride; fazarabine; fenretinid; floxuridine; fludarabina phosphate; fluorouracil; fluorocitabine; foskvidon (Go ^ sspDope); fostricin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmophosin; interleukin II (including recombinant interleukin II, or T1L2), interferon alfa-2a; interferon alfa-2b; interferon alfa-p1; interferon alfa-p3; interferon betaI; interferon gamma b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide (1pD) acetate; liarozola hydrochloride; Lometrexol Sodium; lomustine; losoxantrone hydrochloride; mazoprokol; Maytansin (Tau1ai8she); mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprene (te! orppe); meturedep; mitindomid; mitocarcin; mitocromin (tyoethosh); mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxysuran; pegaspargazu; peliomycin; Pentamustine (RayatikDpe); peplomycin sulfate; perphosphamide; pipobroman; piposulfan (p1ro8i1gap); pyroxantrone hydrochloride; Plykamycin; plomestan; sodium porfimer; porphyromycin; Prednimustin; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprin; rogletimid; safining; safingol hydrochloride; semustin; simtrasen; sparphosate (8Go8a1e) sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin SchgerUpp); streptozocin; sulofenur; talizomycin; tekogalan (1soda1ap) sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; tiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; tricyribin phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozol hydrochloride; uramustine; Uredep; vapreotide; Verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; Vindesine sulfate; vinepidine sulfate; screw glycinate sulfate; Winlerozina sulfate; vinorelbine tartrate; Vinrosidine Sulfate; Vinzolidine Sulfate; Vorosol; zeniplatin; zinostatin; zorubicin hydrochloride. Other anti-cancer agents / drugs that can be used according to the present invention include, without limitation, 20-epi-1.25 dihydroxyvitamin Ό3; 5 ethynyl uracil; abiraterone; aclarubicin; acylfulvene; adeciphenol (desupore1); adozelesin; aldesleukin; ab-TC antagonists; altretamine; ambamustine; amidox (anidox); amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist Ό; antagonist C; antarelix; antidorsal morphogenetic protein-1; antiandrogen prostate carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; afidicolin glycinate; apoptosis gene modulators; regulators of apoptosis; apuric acid; ata-sor-ov-rtva; arginine deaminase; asulacrin (aiyasppe); amestane; atrimustine; Aksinastatin (AhtaChipe) 1; Aksinastatin (Ahsha81a1sh) 2; aksinastatin (ahtaChaOp) 3; azasetron; azatoxin (a / aU.ht); azatirosine; derivatives

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baccatin III; balanol; batimastat; antagonists of the hybrid protein VSC / AB; benzochlorins; benzoylstaurosporin (LSP / OyMaigochrops); beta-lactam derivatives; beta-aletin (le1a-a1e1xie); Betaclamycin B; betulinic acid; LPOR inhibitor (basic fibroblast growth factor); bicalutamide; bisantrene; BisaziridinilSpermin (Uka / 1ptyu1kregtte); bisnafid; bistraten A; bizelesin; breflat (legea! e); bopirimine; budotitan; butionin sulfoximine; calcipotriol; calfostin C; camptothecin derivatives (satruJews); canaripox 1b-2; capecitabine; carboxamide-amino triazole; carboxyamidotriazole; Sakek! M3; SAKI 700; inhibitor derived from cartilage; carzelesin; casein kinase inhibitors (1CO8); castanospermin; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomiphene analogues; clotrimazole; kollismitsin (sOshkshu) A; kollismitsin (sOshkshu) V; combretastatin A4; combretastatin analogue; conagenin (sopadeshp); crambescidin 816; Chrisnatol; cryptophycin (cure!) 8; derivatives of cryptophycin (sgur! orusss) A; Curacin A; cyclopentanthraquinones; cycloplates; Cypemycin (suretussh); cytarabine oxyphosphate; cytolytic factor; cytostatin; dacliximab (yasP.Hipa); decitabine; dehydrodidemne (yeBuigoyetshchi) B; deslorelin; dexamethasone; dexyphosphamide (eHGG); dexrazoxane; dexverapamil; diazikvon (yta / ktsyupe); Didemnin B; didox; diethylnorspermin; dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; droloxifene; Dronabinol; duocarmycin (jyosagtush) FOR; abselen; ecomustine; edelfozin; edrecolomab; eflornithine; elements; Emitfur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; ethanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinid; filgrastim; finasteride; flavopiridol; flelasastin; flousterone (Jiakégoie); fludarabine; fluorodounorubicin hydrochloride; forphenimex; Formestan; fostriecin; fotemustine; gadolinium texafirin; gallium nitrate; halocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; Hepsulfam (Verkias); heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramanton; ilmophosin; ilomastat; imidazoacridon; imiquimod; immunostimulating peptides; insulin-like growth factor 1 receptor inhibitor; interferon agonists; interferons; interleukins; yobenguan; iododoxorubicin (yuohogi oss); 4-ipomelanol (1roshaeio1); iroplact; yrsogladin; Isobenhazol (Eoida / o1e); isohomogalicondrin OkonoshoNaPsopipp) B; itasetron; jasplaquinolide; Kagalalid (Kakayaye) R; lamellarin N triacetate; lanreotide; lynamycin; lenograstim; lentinana sulfate; leptolstatin (1pi1k1ayi); letrozole; leukemia inhibiting factor; alpha interferon leukocytes; leuprolide (1eirgoyye) + estrogen + progesterone; leuprorelin; levamisole; liarozol; analogue of linear polyamines; lipophilic disaccharide peptide; platinum lipophilic compounds; lissoclinamide (IkkosPtpiye) 7; lobaplatin; lombricin; lometrexol; lonidamin; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium toxaphyrin; lysophylline; lytic peptides; maytansine; mannostatin A; marimastat; mazoprokol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; Merbaron; Meterelin (Scheidel); methioninase; metoclopramide; M1P inhibitor; mifepristone; miltefosin; myrimostim; non-complementary double-stranded RNA; mitoguazone; mitolactol; mitomycin analogs; mitonaphid; mitotoxin, fibroblast growth factor-saporin; mitoxantrone; Mofaroten; molgramostim; monoclonal antibody, human chorionic gonadotropin; monophosphoryl lipid A + mycobacterial kk cell walls; mopidamol; inhibitor of genes responsible for multidrug resistance; therapy based on suppressor gene 1 multiple tumors; mustard anti-cancer agent; mikaperoksid (shusaregokh) V; cell wall extract of mycobacteria; Miriaporon (foolish); Ацет-acetyldinaline; Ν-substituted benzamides; nafarelin; nagrestip (iadgekyr); naloxone + pentazocine; napavin; nafterpin (IarIegrsch); nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin (shkalshusssh); nitric oxide modulators; nitroxide antioxidant; nitrulline; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracine; oral cytokine stimulator; ormaplatin; ozateron; oxaliplatin; oxaunomycin; Palauamine (Pa); palmitoylrisoxin; pamidronic acid; panaxitriol; panomifene; parabaktin; paselliptin; pegaspargaz; peldesin; pentosan sodium polysulfate; pentostatin; pentrozole; perflubron; perphosphamide; perillyl alcohol; Phenazinomycin (pHet / Totust); phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; pyriterexim; placentin (p1aseyi) A; placentin (p1aseyi) B; plasminogen activator inhibitor; platinum complex; platinum compounds; triamine platinum complex; sodium porfimer; porphyromycin; prednisone; propyl bis-acridone; prostaglandin 12; proteasome inhibitors; an immune modulator based on protein A; protein kinase C inhibitor; protein kinase C inhibitors, microalgal (sh1sgoa1da1); protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurines; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; KaGbolkov antagonists; raltitrexed; ramosetron; inhibitors of farnesyltransferase, disrupting the function of proteins How; protein inhibitors Like; an inhibitor of As-OAP; demethylated retelliptin; rhenium Ke 186 etidronate; Risoxin (G1I / OHT); ribozymes; K11 retinamide; rogletimid; Rohitukin; romurtis; roquinimex; rubiginone B1; ruboxyl; safining; saintopin (cashmere); FSII; sarcophitol A; sargrammost; mimetics zy | 1 (Keieksei! Yeyuyy Shyyog 1); semustin; aging inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen - 9 029372

binding protein; sizofiran; Sobuzoxan; sodium borocaptate; sodium phenylacetate; solverol (8-11); somatomedin binding protein; sonermine; sparphosic acid; spicamycin Ό; spiromustine; splenopentin; Spongistatin (SropdSchip) 1; squalamine; stem cell inhibitor; stem cell division inhibitors; stipiamide (Γΐίρίαιηίώ;); stromelysin inhibitors; sulphinozin (iosixio); superactive antagonist vasoactive intestinal peptide; suradistu (8igab181a); suramin; svainsonin; synthetic glycosaminoglycans; tallimustine; tamoxifen metiodide; tauromustine; tazaroten; tekogalan (1eoda1a) sodium; tegafur; tellurium (1E11igarugu1sht); telomerase inhibitors; temoporfin; temozolomide; teniposide; carbon tetrachloride; tetrazomine (1ela / oti); Taliblastin (Shapykie); tiocoraline; thrombopoietin; thrombopoietin mimetic; thymalphasin; thymopoietin receptor agonist; timotrinan; thyroid stimulating hormone; ethyl ethiopurpurin tin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; tricyribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; inhibitors of ICS; umenimex; inhibitory growth factor secreted by the urogenital sinus; urokinase receptor antagonists; vapreotide; Variolin B; gene therapy of erythrocyte diseases using vector systems; Velaresol; veramine (ugatta); Verdins; Verteporfin; vinorelbine; Winxaltine (WighaUpe); Vitaxin; Vorosol; Zanoterone; zeniplatin; zilaskorb and zinostatin stimalamer. Preferred additional anti-cancer drugs are 5-fluorouracil and leucovorin (first-generation).

According to one embodiment of the present invention, anti-cancer agents that can be used in the methods described in this invention are selected from the group consisting of paclitaxel, docetaxel, 5-fluorouracil, trastuzumab, lapatinib, bevacizumab, letrozole, goserelin, tamoxifen, cetuximab, panitumumab, gemcitabine, capecitabine, irinotecan, oxaliplatin, carboplatin, cisplatin, doxorubicin, epirubicin, cyclophosphamide, methotrexate, vinblastine, vincristine, melphalan, and combinations thereof.

According to one embodiment of the present invention, the anti-cancer agent and the compound represented by the structural formula (I) are administered simultaneously. When administered simultaneously, the anticancer agent and the compound can be administered as part of the same preparation or as part of different preparations. Alternatively, the compound and the additional anti-cancer agent are administered separately. Alternatively, the compound and the additional anti-cancer agent can be administered sequentially, as separate compositions, during a suitable time interval (for example, during the course / period of cancer treatment (for example, from about 1.5 hours to about 5 hours, or approximately 10 hours, or up to approximately 15 hours, or up to approximately 20 hours, from approximately 1 to approximately 2 days, or up to approximately 5 days, or up to approximately 10 days, or up to approximately 14 days (), which is determined by the experienced clinical rach (for example, for a period of time sufficient for the pharmaceutical action of the two types of therapy to overlap). The compound and the additional anti-cancer agent can be administered as a single dose or as multiple doses using a sequence and administration scheme suitable for achieving the desired therapeutic effect (for example, inhibiting tumor growth).

According to one embodiment of the present invention, in the methods described in the present invention, the subject has not previously received treatment with a protein kinase inhibitor TTC (for example, a compound represented by the structural formula (I)).

The term "inhibition of growth of cells initiating tumor development" refers to a decrease in the rate of proliferation and / or survival of cells initiating the development of a tumor.

In the context of this description, the term "reducing the likelihood of cancer recurrence" means partial or complete inhibition, slowing the recurrence of cancer in the place of primary localization, or next to it, and / or in the place of secondary localization after a period of remission. This term also means that the likelihood of cancer re-developing after the treatment described in this document is less than in the absence of this treatment.

In the context of this description, the term "remission" refers to a cancer condition in which the clinical symptoms or indicators associated with cancer disappear or cannot be registered, usually after the subject has been successfully treated with anticancer therapy.

In the context of this description, "treatment" is an approach for obtaining favorable or desired results, including clinical results. Favorable or desired clinical results may include, without limitation, alleviating or reducing the severity of one or more symptoms, or alleviating or improving the condition, reducing the extent of the disease, stabilizing (that is, not worsening) the course of the disease, reducing the likelihood of spreading the disease, delaying or slowing the progression of the disease, improvement or temporary relief of a painful condition, registered or unregistered. "Treatment" can also mean an increase in survival compared to the expected survival without treatment. "Treatment" also includes reducing the likelihood of disease recurrence.

- 10 029372

In the context of this description, “treating a subject with cancer” includes achieving, in part or substantial, one or more of the following results: cessation of development, reduction in the incidence of cancer (for example, reduction in tumor size), inhibition of the rate of cancer development, decrease in intensity or an improvement in a clinical symptom or indicator associated with cancer (such as tissue or serum components), or an increase in the lifespan of the subject and a decrease in the likelihood of cidiva cancer.

Typically, an effective amount of a compound according to the present invention varies depending on various factors, such as the drug or compound administered, the pharmaceutical preparation, the route of administration, the type of disease or disorder, the individual characteristics of the subject or organism being treated, and the like, but may nevertheless be determined by a person skilled in the art in the usual way. An effective amount of a compound according to the present invention can be easily determined by one of ordinary skill in the art using standard techniques known in the art.

The term "effective amount" means an amount that, when administered to a subject, leads to favorable or desired results, including clinical results, for example, inhibits, stops or reduces the spread of cancer (for example, according to clinical symptoms or cancer cells) in a subject compared to control.

According to one embodiments of the invention, the effective amount of the compounds according to the present invention ranges from about 0.1 to about 1000 mg / kg body weight, alternatively, from about 1 to about 500 mg / kg body weight and, alternatively, from about 20 up to about 300 mg / kg body weight. According to another embodiment of the invention, an effective amount of a compound according to the present invention ranges from about 0.5 to about 5000 mg / m, alternatively from about 5 to about 2500 mg / m and, alternatively, from about 50 to about 1000 mg / m . The specialist in this field it is clear that some factors may influence the amount of dose required for effective treatment of a subject suffering from cancer, or reduce the likelihood of recurrence of cancer. These factors include the severity of the disease or disorder, previous treatment, general health and / or age of the subject, and other diseases of the subject, but are not limited to them.

In addition, in the methods described in the present invention (including treating a subject suffering from cancer, or reducing the likelihood of cancer recurring), the subject’s “treatment” or dosage regimen to the subject using an effective amount of a compound according to the present invention may consist of a single injection or alternatively include a series of introductions. For example, the compound of the present invention may be administered at least once a week. However, according to another embodiment of the present invention, a specific treatment implies that the compound can be administered to a subject from about once a week to once a day. The duration of treatment depends on a number of factors, such as the severity of the disease, the patient's age, the concentration and activity of the compounds according to the present invention, or a combination thereof. It is also necessary to take into account that the effective dose of the compound used for treatment may be increased or decreased during the course of treatment using a specific treatment regimen. The need to change the dose may become apparent after analyzing the data obtained when performing standard diagnostic tests known in the art. In some embodiments, a prolonged administration may be required.

A "subject" is a mammal, preferably a human, but may also be an animal that needs veterinary care, such as domestic animals (eg, dogs, cats, etc.), farm animals (eg, cattle, sheep, pigs , horses, etc.) and laboratory animals (eg, rats, mice, guinea pigs, etc.).

The compounds of the present invention can be administered to a patient in various forms, depending on the chosen route of administration, as will be understood by those skilled in the art. The compounds of the present invention can be administered, for example, by the oral, parenteral, buccal, sublingual, nasal, rectal route, using a patch, a spray or transdermal administration, and in the form of pharmaceutical compositions intended for the respective administration. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and local routes of administration. Parenteral administration may be a continuous infusion over a fixed period of time.

The compounds of the present invention may be presented in the form of appropriately prepared pharmaceutical compositions intended for administration to a subject. The pharmaceutical compositions of the present invention optionally comprise one or more pharmaceutically acceptable carriers and / or diluents, such as lactose, starch, cellulose, and dextrose. Other excipients, such as flavoring agents, sweeteners and preservatives, such as methyl, ethyl, propyl and butyl parabens, may also be included. A more complete list of suitable

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excipients can be found in "Napbook οί RNagtaseibsa1 ΕχοίρίοηΙκ" (5 ^'1' Eb., RNagtaseibsa1 Rgekk (2005). One skilled in the art knows how to obtain preparations suitable for various administration routes. The standard techniques and ingredients for the selection and preparation suitable drugs are described, for example, in “Retponschüp'n RNagtseiis1 Zaeisek” (2003, 20 ^{1 1} еб ebShoi) and in “Thé Iieeb 81aRek Rnagtasore1a: THnΝ ΝΡ; · ιΙίοη; · ι1 Horns” (R8r 1999); Carriers, diluents and / or excipients are “acceptable” in the sense of being compatible with the other ingredients. ntami pharmaceutical composition and a sense of safety to the recipient thereof.

Typically, a compound of the present invention intended for oral therapeutic administration can be mixed with an excipient and administered in the form of swallowable tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, and the like.

Generally, solutions of a compound of the present invention intended for parenteral administration can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol and in oils. These preparations intended for storage and use in standard conditions contain a preservative to prevent the proliferation of microorganisms.

Generally, sterile aqueous solutions or dispersions of a compound according to the present invention and sterile powders of a compound according to the present invention suitable for extemporally preparing sterile injection solutions or dispersions are suitable for injection use.

Compounds according to the present invention, intended for nasal administration, can be represented in the form of the drug in the form of aerosols, drops, gels and powders. Aerosol preparations usually contain a solution or a highly dispersed suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually present in an amount corresponding to one dose or several doses, in sterile form, in a hermetically sealed container, which can be in the form of a cartridge or a reusable container designed for use with a spray device. Alternatively, the hermetically sealed container may be a single charge dispenser, such as a single dose nasal inhaler or an aerosol dispenser equipped with a metering valve that is discharged after use. When the dosage form comprises an aerosol dispenser, it usually contains a propellant, which may be a compressed gas, such as compressed air, or an organic propellant, such as a fluorochlorine-substituted hydrocarbon. Aerosol dosage forms can also be presented in the form of a spray with a pump.

Compounds of the present invention intended for buccal or sublingual administration may be presented as a formulation containing a carrier such as sugar, gum arabic, tragacanth or gelatin and glycerin, in the form of tablets, lozenges or lozenges.

Compounds of the present invention intended for rectal administration may be presented as a preparation in the form of suppositories containing a standard suppository base such as cocoa butter.

Compounds according to the present invention can be obtained using methods known to experts in this field of technology, as illustrated by the general schemes and techniques and the preparative examples described below. All starting materials are either commercially available or prepared using techniques known to those skilled in the art and those described below.

In accordance with another aspect of the present invention, the compounds according to the invention can be obtained using methods similar to those known in the art. General methods for the synthesis of compounds described in the claims, are detailed below in example A.

Examples

Example A. Synthesis.

General techniques.

Commercially available starting materials, reagents and solvents used as is. In general, anhydrous reactions were carried out under an atmosphere of an inert gas, such as nitrogen or argon. Commercially available RogaRak® Khi CX resin was used as a cation exchange resin.

Reactions using microwave radiation were carried out in a Vyuade 1p111-1og microwave reactor. Over the course of the reaction is usually monitored using TLC (thin layer chromatography), performed on silica gel plates (Megsk), with visualization of UV radiation at a wavelength of 254 nm, as well as using analytical HPLC or LC-MS (Vigciege 4000 or system \ Wileck Get good for IRBS (super-performance liquid chromatography)). Purification of intermediate compounds or final products was performed using flash column chromatography on silica gel 60 (230-400 mesh) from EMI CNENISK or 8CHus1e, or using the Vude 1cotega system on silica gel cartridges KP-81b or HP-81b or mainly modified 12.029372

ΚΡ-ΝΗ silica bath and cartridges. Purification using reverse-phase HPLC was performed on a Wapai Rter81at Model 8Ό-1 HPLC system equipped with a Wapai Moytot 10 Phase-18 C-18 back-phase column using approximately 5-30% MeCI, or MeOH / 0.05% TFC-H ₂ About 70-90% MeSI, or MeOH / 0.05% TFA in H ₂ O for 20-40 min at a flow rate of 30-80 ml / min. Purification using reverse phase chromatography was also performed on a VyuLaGe1Go1eta system equipped with an KR-C18-H column using 10-95% MeOH or CH ₃ SI / 0.1% TFA in H ₂ O. Proton NMR spectrum A 400 MHz spectrometer was recorded on a spectrometer and mass spectra were obtained using a Vickkeg E ^ spectral spectrometer 4000 or an IRBC system of Hailet8 LSRSch.

The names of the compounds are generated using the program embedded in SATpbgse8oyRetkte1tet'8 S'etVyuEg y UIt, version 11.0 or 12.0.

In the description used the following abbreviations and abbreviations: water. - water;

Ar - argon;

Vos - tert-butoxycarbonyl;

Yr - broad; sa1sb - calculated; b - doublet;

DHM - dichloromethane;

ΌΙΡΕΑ - diisopropylethylamine;

DMF - Ν,-dimethylformamide;

DMSO - dimethyl sulfoxide;

BrrH - 1,1'-bis- (diphenylphosphino) ferrocene;

h - hour (s);

HPLC — High Performance Liquid Chromatography;

LC-MS — liquid chromatography / mass spectrometry; min - minute (s);

t - multiplet;

MS-IER - mass spectrometry with electrospray ionization;

NMR - nuclear magnetic resonance;

Ο / Ν - during the night;

PE - petroleum ether;

RMB is para-methoxybenzyl;

rger - preparative;

c.t. - room temperature;

8 - singlet;

ί - triplet;

TFA - trifluoroacetic acid;

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Intermediate compounds

4-Bromo-X-cyclopropyl-2-methylbenzamide

To a suspension of 4-bromo-2-methylbenzoic acid (43.0 g, 200 mmol) and oxalyl dichloride (30.5 g, 240 mmol) in DCM (300 ml) was added DMF (0.1 ml). The resulting reaction mixture was stirred for 16 hours at room temperature. Over the course of 16 hours, the reaction mixture slowly turned into a clear yellow solution. Then the solvent was removed under vacuum and this crude product was used in the next step without further purification. The resulting crude product was redissolved in DCM (300 ml) and cooled to 0 ° C. A mixture of TEA (42 ml, 300 mmol) and cyclopropylamine (12.6 g, 220 mmol) in DCM (100 ml) was added slowly over 15 minutes and the mixture was stirred for 2 hours at room temperature. Then the reaction mixture was diluted with DCM (200 ml) and water was added. The resulting mixture was extracted with DCM and the combined organic extracts were dried over MD80 ₄ and concentrated to yield the desired product as a pale pink solid (50.1 g, 99%).

¹ H NMR (400 MHz, COC1 ₃ ) δ ppm: 7.37 (§, 1H), 7.31 (ά,> 8.0 Hz, 1H), 7.17 (, 1 = 8.1 Hz, 1H), 5.93 (Zr. §, 1H), 2.90-2.85 (t, 1H), 2.40 (§, 3H), 0.90-0.85 (t, 2H), 0.62-0.58 (t, 2H);

MS-IER: [M + H] + 253.9; calculated for [С ₁₁ Н ₁₂ ВгК0 + Н] + 254.0.

X- (4-Bromo-2-methylphenyl) cyclopropanecarboxamide

In 100 ml of HGV, 4-bromo-2-methylaniline (3.7 g, 20 mmol) and (6.95 ml, 40 mmol) were mixed with DMF (40 ml). This reaction mixture was cooled to 0 ° C in an ice bath and the acid chloride (2.1 g, 20 mmol) was added to cyclopropanecarboxylic acid. The resulting mixture was stirred for 1 h at 0 ° C. Water and EtOAc (ethyl acetate) were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The organic layers were combined, dried over α ₂ 80 ₄ , filtered and concentrated under reduced pressure to give the title compound as a white solid (4.76 g, 94%).

¹ H NMR (400 MHz, COCl ₃ ) δ ppm: 7.85-7.72 (t, 1H), 7.38-7.28 (t, 2H), 7.15-7.02 (t, 1H), 2.27 (§, 3H), 1.57-1.48 (t, 1H), 1.10 (ςυίηί, 1 = 3.9 Hz, 2H), 0.92-0.79 (t, 2H);

MS-IER: [M + H] + 253.9; calculated for [С ₁₁ Н ₁₂ ВгК0 + Н] + 254.0.

X-Cyclopropyl-2-methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl) benzamide

A mixture of 4-bromo-X-cyclopropyl-2-methylbenzamide (3.73 g, 14 mmol), bis- (pinacolato) dibor (5.59 g, 22 mmol) and K0Ac (4.29 g, 43 mmol) in DMF (37 ml) was purged with argon for 10 minutes at room temperature. RbCl ₂ (brpl) -DXM (0.59 g, 5 mol%) was then added and the reaction mixture was heated at 100 ° C in an oil bath for 4 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (200 ml) and water (100 ml). The combined layers were filtered through a pad of diatomaceous earth (Ceillet) and the pad was washed with a small amount of EATA. The aqueous layer was further extracted with ethyl acetate (50 ml) and the combined organic layer was washed with brine, dried over Να ₂ 80 ₄ , filtered and concentrated to give the crude oil as a residue. The resulting crude product was purified by flash chromatography (gradient: EUT / hexane 0100%) to yield the title compound as a cream-colored solid (4.15 g, 94%).

¹ H NMR (400 MHz, COC1 ₃ ) δ ppm: 7.66 (§, 1Н), 7.63-7.61 (ά,> 7.6 Hz, 1 N), 7.32-7.30 (,> 7.6 Hz, 1H), 5.85 (§, 1 Н), 2.93-2.87 (t, 1Н), 2.46 ( ₈ , 3Н), 1.35 ( ₈ , 12Н), 0.90-0.86 (t, 2Н), 0.63-0.59 (t, 2Н);

MS-IER: [M + H] + 302.2; calculated for [C _p H ₂₄ VX0 ₃ + H] + 302.2.

3-Bromo-5,7-dichloropyrazolo [1,5-a] pyrimidine

To a stirred solution of sodium ethoxide in EUN, which was obtained from sodium (281.3 g,

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12.0 mol) and ΕίΘΗ (10 l) using standard procedures, malonic acid diethyl ether (963.7 g, 6.02 mol) was added at ambient temperature and then the compound 1H-pyrazole-3amine (500 g, 6, 02 mole). The reaction mixture was refluxed for 12 hours. After cooling to room temperature, the precipitate was collected by filtration and dissolved in water. The resulting aqueous solution was acidified by the addition of 2 M HC1 (pH 2). The resulting precipitate was collected by filtration and dried under reduced pressure to give pyrazolo [1,5-a] pyrimidine 5.7 (4H, 6H) -dione (649 g, 71%) as a yellow solid, which was used in the next step without additional cleaning up.

A stirred suspension of pyrazolo [1,5-a] pyrimidine-5,7 (4H, 6H) -dione (265 g, 1.75 mol) and Ν,-dimethylaniline (335.6 ml) in POC1 ₃ (2.00 kg, 13.2 mol) was refluxed for 4 hours. After cooling to room temperature, the reaction mixture was poured into ice water and stirred for 30 minutes, neutralized by the addition of a saturated aqueous solution of sodium carbonate and extracted with ethyl acetate. The combined organic layers were washed with water, brine and dried over Μ β δ ₄ , filtered and evaporated. The residue was purified by silica gel column chromatography (gradient: EUAc / PE 1:10) to give 5,7-dichloropyrazolo [1,5-a] pyrimidine (287 g, 87%) as a yellow solid. To a solution of 5,7-dichloropyrazolo [1,5-a] pyrimidine (246.6 g, 1.31 mol) in ΟΠΥΝ (1.8 l) was added ΝΒδ (-bromosuccinimide) (245 g, 1.38 mol) . The resulting mixture was stirred for 2 hours at room temperature. After removing the solvent, the reaction mixture was purified by silica gel column chromatography (gradient: E1OAc / PE 1: 5) to obtain the title compound (313.5 g, 89%) as a light yellow solid.

¹ H NMR (300 MHz, COCl ₃ ) δ ppm: 7.04 (δ, 1H), 8.21 (δ, 1H);

MS-IER: [M + H] + 265.9; calculated for [С ₆ Н ₂ ВгС1 ₂ Н ₃ + Н] + 265.9.

3 -Brom-5-chloro-Y- (cyclopropylmethyl) pyrazolo [1,5-a] pyrimidine-7-amine

To a solution of 3-bromo-5,7-dichloropyrazolo [1,5-a] pyrimidine (5.0 g, 19 mmol) in DCM (50 ml) was added cyclopropylmethanamine (1.48 g, 21 mmol) and ΏΙΡΕΑ (6, 6 ml, 38 mmol). This reaction mixture was stirred for 2 hours at room temperature. Water and DCM were added, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic extracts were dried over # 8O ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUT / hexane 0-50%) to yield the title compound as a yellow solid (5.25 g, 92%).

¹ H NMR (400 MHz, COC1 ₃ ) δ ppm: 7.97 (δ, 1H), 6.50 (Lg. Δ, 1H), 5.97 (δ, 1H), 3.25 (, 1 = 7.3, 5.5 Hz, 2H), 1.26-1.13 (t, 1H), 0.74-0.65 (t, 2H), 0.37 (c, 1 = 5.0 Hz, 2H);

MS-IER: [M + H] + 301.0; calculated for | C | ₀ 1 Ι | ₀ Βΐ '(Ί \ ι · 1 |' 301.0.

(1δ, 3δ) -3 - (((3 -Brom-5-chloropyrazolo [1,5-a] pyrimidin-7-yl) amino) methyl) -1-methylcyclobutanol

To a solution of 3-bromo-5,7-dichloropyrazolo [1,5-a] pyrimidine (14.6 g, 55.3 mmol) in DCM (200 ml) was added cis-hydroxy-3-methylcyclobutan-1-methylamine (7 , 0 g, 60.9 mmol) and ΏΙΡΕΑ (19.2 ml, 110.6 mmol). This reaction mixture was stirred for 16 hours at room temperature. Water and DCM were added, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic extracts were dried over No. ₂ δΟ ₄ , filtered, and concentrated to give the title compound as a yellow solid (18.1 g, 95%).

¹ H NMR (400 MHz, CoC1 ₃ ) δ ppm: 7.96 (δ, 1H), 6.60-6.49 (t, 1H), 5.99 (δ, 1H), 3.47 (1, 1 = 6.0 Hz, 2H) , 2.38-2.27 (t, 3H), 1.96-1.85 (t, 2H), 1.43 (δ, 3H);

MS-IER: [M + H] + 345.1; calculated for [C1 ₂ H ₁₄ VgSSh ₄ O ₂ + H] + 345.0.

tert-Butyl (3-bromo-5-chloropyrazolo [1,5-a] pyrimidin-7-yl) (cyclopropylmethyl) carbamate

To a solution of 3-bromo-5-chloro-L- (cyclopropylmethyl) pyrazolo [1,5-a] pyrimidine-7-amine (5.25 g, 17.5 mmol) in DCM (150 ml) was added Voc ₂ O ( 5.70 g, 26.2 mmol), (4-dimethylaminopyridine)

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(0.21 g, 1.75 mmol) and TEA (7.3 ml, 52.5 mmol). This reaction mixture was stirred for 4 hours at room temperature. Water and DCM were added, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic extracts were dried over ΜβδΟ ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUTC / hexanes 0-40%) to yield the title compound as a yellow solid (6.24 g, 89%).

¹ H NMR (400 MHz, SESl3) δ ppm: 8.12 (δ, 1H), 6.85 (δ, 1H), 3.72 (, 1 = 7.3 Hz, 2H), 1.39 (δ, 9H), 1.050. 94 (t, 1H), 0.46-0.37 (t, 2H), 0.13-0.03 (t, 2H);

MS-IER: [M-С ₄ H ₈ ] + 345.0; calculated for [С15Н18ВгС1Х4О2-С4Н8] ⁺ 345.0.

tert-Butyl (3-bromo-5-chloropyrazolo [1,5-a] pyrimidin-7 -yl) ((((1δ, 3δ) -3 - ((tert-butoxycarbonyl) oxy) -3-methylcyclobutyl) methyl) carbamate

To a solution of (1δ, 3δ) -3 - (((3-bromo-5-chlorpyrazolo [1,5-a] pyrimidine-7 -yl) amino) methyl) -1 methylcyclobutanol (18.1 g, 52.6 mmol ), Boc ₂ O (34.3 g, 158 mmol) and TEA (22 ml, 157.8 mmol) in DCM (200 ml) were added with OMAP (1.28 g, 10.5 mmol). This reaction mixture was stirred for 16 hours at 40 ° C. The solvent was removed under vacuum and water and DCM were added, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic extracts were dried over a5O ₄ , filtered, and concentrated. The resulting crude product was purified by flash chromatography (simultaneously on 3 columns of Bülage (100 g MO ₂ ); gradient: EALAc / hexanes 0-30%) with the yield of the title compound as a beige solid (12.7 g, 44%).

¹ H NMR (400 MHz, COCl ₃ ) δ ppm: 8.11 (δ, 1H), 6.74 (δ, 1H), 3.88 (ά, 1 = 6.5 Hz, 2H), 2.17 (δ, 3H), 1.991 .88 (t, 2H), 1.47 (δ, 3H), 1.45 (δ, 9H), 1.35 (δ, 9H);

MS-IER: [M + H] + 545.1; calculated for [С ₂₂ Н ₃₀ ВгС1 ₄ О ₅ + Н] + 545.1.

(1δ, 3δ) -3 - (((3 -Brom-5-chlorpyrazolo [1,5-a] pyrimidin-7-yl) (4-methoxybenzyl) amino) methyl) -1 methylcyclobutanol

To a solution of (1δ, 3δ) -3 - (((3-bromo-5-chlorpyrazolo [1,5-a] pyrimidin-7-yl) amino) methyl) -1 methylcyclobutanol (12.0 g, 34.7 mmol ) and 4-methoxybenzyl chloride (5.2 ml, 38.2 mmol) in DMF (50 ml) was added K ₂ CO ₃ (9.6 g, 69.4 mmol). The resulting mixture was stirred for 3 hours at 60 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were washed with water, dried over MgSO ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUTC / hexanes 10-100%) using three columns simultaneously. The fractions containing impurities were collected and purified by flash chromatography using the aforementioned purification conditions. The fractions containing pure product were combined and concentrated to give the desired product as a pale yellow solid (13.1 g, 81%).

¹ H NMR (400 MHz, CO13) δ ppm: 8.01 (δ, 1H), 7.17 (ά, 1 = 8.7 Hz, 2H), 6.67 (, 1 = 8.7 Hz, 2H), 6.02 (δ, 1H), 4.98 (δ, 2H), 3.83 (ά, 1 = 6.4 Hz, 2H), 3.81 (δ, 3H), 2.25-2.16 (t, 3H), 1.76-1.72 (t, 2H), 1.36 (δ , 3H);

MS-IER: [M + H] + 467.1; calculated for [С ₂₀ Н ₂₂ ВгСШ ₄ О ₂ + Н] + 467.1.

tert-Butyl (3-bromo-5 - (((18.2K) -2-hydroxycyclohexyl) amino) pyrazolo [1,5-a] pyrimidine-7yl) (cyclopropylmethyl) carbamate

To a solution of tert-butyl (3-bromo-5-chloropyrazolo [1,5-a] pyrimidine-7yl) (cyclopropylmethyl) carbamate (9.0 g, 22.5 mmol) in (90 ml) was added (1Κ, 2δ ) -2-aminocyclohexanol-HC1 (4.08 g, 27.0 mmol) and O1REA (3.47 g, 25.1 mmol). This reaction mixture was divided into six portions, each of which was placed in a tightly closed tube for a microwave oven. Each tube was heated in a microwave for 3 hours at 130 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over No. ₂ 8O ₄ , filtered and concentrated. The resulting crude

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the product was purified by flash chromatography (gradient: EALA / hexanes 0-50%) using three columns simultaneously, with the release of the title compound as a beige solid (8.51 g, 79%).

Υ NMR (400 MHz, COClS) δ ppm: 7.80 (δ, 1H), 6.08 (δ, 1H), 5.33-5.19 (t, 1H), 4.26-4.14 (t, 1H), 4.134.06 ( t, 1H), 3.61 (V, 1 = 7.3 Hz, 2H), 2.51 (lg. δ, 1n), 1.89-1.62 (t, 8H), 1.55-1.45 (t, 2H), 1.40 (δ, 9H) , 1.060.95 (t, 1H), 0.47-0.38 (t, 2H), 0.17-0.07 (t, 2H);

MS-IER: [M + H] + 408.3; calculated for [С ₂ 1Н30ВгЫ5О3 + Н] ⁺ 480.2.

tert-Butyl (3-bromo-5- (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidin-7-yl) (cyclopropylmethyl) carbamate

A mixture of tert-butyl (3-bromo-5-chloropyrazolo [1,5-a] pyrimidin-7-yl) (cyclopropylmethyl) carbamate (400 mg, 1.0 mmol), 3-hydroxypyridine (475 mg, 5.0 mmol ), ΌΒυ (0.75 ml, 5.0 mmol) in DME (dimethoxyethane) (5 ml) were placed in a tightly closed tube for a microwave oven. The mixture was heated in a microwave for 1 hour at 85 ° C. Water and EA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EALA / hexanes 0-50%) to yield the title compound as a brown oil (367 mg, 80%).

¹ H NMR (400 MHz, SPS) δ ppm: 8.64 (, 1 = 2.5 Hz, 1H) 8.54 (άά, 1 = 4.5, 1.5 Hz, 1H) 7.97 (δ, 1H), 7.787.72 ( t, 1H), 7.44-7.38 (t, 1H) 6.61 (δ, 1H) 3.71 (ά, 1 = 7.3 Hz, 2H) 1.39 (δ, 9H), 1.06-1.01 (t, 1H) 0.45 (δ, 2H), 0.13-0.09 (t, 2H);

MS-IER: [M + H] + 460.0; calculated for [С ₂ 0Н ₂₂ ВГЫ5О3 + Н] + 460.1.

tert-Butyl (3-bromo-5- (cyclopentylamino) pyrazolo [1,5-a] pyrimidin-7-yl) (((18.38) -3 - ((tertbutoxycarbonyl) oxy) -3-methylcyclobutyl) methyl) carbamate

To a solution of tert-butyl (3-bromo-5-chloropyrazolo [1,5-a] pyrimidin-7-yl) (((E ^) - 3 - ((tertbutoxycarbonyl) oxy) -3-methylcyclobutyl) methyl) carbamate ( 0.45 g, 0.83 mmol) in (4 ml) was added cyclopentylamine (0.085 g, 1.0 mmol) and GIREA (0.21 g, 1.66 mmol). This reaction mixture was heated in a microwave for 3 hours at 130 ° C. Water and EA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over Ν ₂ 3Ο ₄ , filtered, and concentrated. The resulting crude product was purified by flash chromatography (gradient: EAU / hexanes 0-60%) to yield the title compound as a white solid (0.40 g, 67%).

¹ H NMR (400 MHz, CBC1 ₃ ) δ ppm: 7.79 (δ, 1H), 5.94 (δ, 1H), 5.08 (Lg. Δ, 1H), 4.23 (Lg. Δ, 1H), 3.85- 3.72 (t, 2H), 2.28-2.06 (t, 5H), 2.02-1.89 (t, 2H), 1.81-1.61 (t, 5H), 1.48 (δ, 4H), 1.44 (δ, 9H), 1.36 ( δ, 9H);

MS-IER: [M + H] + 594.3; calculated for [С ₂₇ Н ₄₀ Вг ^ О ₅ + Н] + 594.2.

tert-Butyl (3-bromo-5 - (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidin-7 -yl) (((1δ, 3 δ) -3 - ((tertbutoxycarbonyl) oxy) -3 -methylcyclobutyl) methyl) carbamate

A mixture of tert-butyl (3-bromo-5-chlorpyrazolo [1,5-a] pyrimidin-7 -yl) (((1δ, 3 δ) -3 - ((tertbutoxycarbonyl) oxy) -3-methylcyclobutyl) methyl) carbamate (12.7 g, 23.3 mmol), 3-hydroxypyridine (4.43 g, 46.6 mmol) and K ₂ CO ₃ (9.65 g, 69.9 mmol) in DMF (100 ml) were stirred in overnight at room temperature. Then the reaction mixture was diluted with ethyl acetate and washed with water, dried over SO ₂ No. ₂ , filtered and concentrated under vacuum. This crude product was triturated with E1 ₂ O and filtered to give the title compound as a beige solid (8.43 g, 14.0 mmol). The mother liquor was concentrated under vacuum and purified by flash chromatography (gradient: EALA / hexane 0-40%) to give the title compound in

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a light orange solid (4.07 g, 6.74 mmol). The solid obtained by trituration and the solid obtained by flash chromatography were combined to obtain the title compound as a light orange solid (12.50 g, 89%).

¹ H NMR (400 MHz, COC1 ₃ ) δ ppm: 8.65 (, 1 = 2.8 Hz, 1H), 8.55 (άά, 1 = 4.6, 1.4 Hz, 1H), 7.98 (δ, 1H), 7.787 .73 (t, 1H), 7.46-7.40 (t, 1H), 6.50 (δ, 1H), 3.89 (, 1 = 6.8 Hz, 2H), 2.30-2.15 (t, 3H), 2.02-1.93 (t , 2H), 1.50 (δ, 3H), 1.45 (δ, 9H), 1.38 (δ, 9H);

MS-IER: [M + H] + 604.3; calculated for [С ₂₇ Н ₃₄ ВгЫ ₅ О ₆ + Н] + 604.2.

(P ^) - 3 - (((3-Bromo-5- (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidin-7-yl) (4methoxybenzyl) amino) methyl) -1-methylcyclobutanol

To a solution of (P ^) - 3 - (((3-bromo-5-chloropyrazolo [1,5-a] pyrimidin-7-yl) (4-methoxybenzyl) amino) methyl) -1-methylcyclobutanol (13.1 g , 28.2 mmol) and 3-hydroxypyridine (4.0 g, 42.3 mmol) in DMF (70 ml) were added TO ₂ CO ₃ (7.8 g, 56.3 mmol). The resulting mixture was stirred for 22 hours at 80 ° C. A mixture of 60% NaH (0.6 g, 15 mmol) and 3-hydroxypyridine (1.4 g, 14.7 mmol) in DMF was stirred for 30 minutes at room temperature and then added to the reaction mixture. The resulting solution was stirred for 20 hours at 100 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were washed with water, dried over MgSO ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUTC / hexanes 10-100%) using three columns simultaneously. The fractions containing impurities were collected and purified by flash chromatography (gradient: EUTC / hexanes 50100%). The fractions containing pure product were combined and concentrated to give the desired product as a pale yellow solid (9.5 g, 64%).

¹ H NMR (400 MHz, COCl ₃ ) δ ppm: 8.56 (ά, 1 = 4 Hz, 1H), 8.45 (ά, 1 = 8 Hz, 1H), 8.00 (δ, 1H), 7.69-7.65 (t, 1H), 7.36 (άά, 1 = 4.8, 0.4 Hz, 1H), 7.19 (ά, 1 = 8.7 Hz, 2H), 6.85 (ά, 7 = 8.7 Hz, 2H), 5.71 (δ, 1H) , 4.91 (δ, 2H), 3.87 (ά, 1 = 6.7 Hz, 2H), 3.87 (δ, 3H), 2.30-2.15 (t, 3H), 1.77 (, 1 = 8.1 Hz, 2H), 1.37 ( δ, 3H);

MS-IER: [M + H] + 524.2; calculated for [С ₂₅ Н ₂₆ ВгЫ ₅ О ₃ + Н] + 524.1.

N-Cyclopropyl-4- (7 - ((((P ^) - 3-hydroxy-3-methylcyclobutyl) methyl) (4-methoxybenzyl) amino) -5 (pyridin-3-yloxy) pyrazolo [1,5-a ] pyrimidine-3-yl) -2-methylbenzamide

A mixture of (P ^) - 3 - (((3-bromo-5- (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidin-7-yl) (4-methoxybenzyl) amino) methyl) -1- methylcyclobutanol (9.5 g, 18.1 mmol), N-cyclopropyl-2-methyl-4- (4,4,5,5-tetramethyl1,3,2-dioxaboran-2-yl) benzamide (7.6 g , 25.4 mmol), 2 M K ₃ PO ₄ (23 ml, 45.3 mmol) in THF (100 ml) were purged with argon for 10 min, then PάC1 ₂ άρρ DHM (1.5 g, 1.8 mmol ). The resulting mixture was heated at reflux in an oil bath for 16 hours at 84 ° C. Then the reaction mixture was diluted with dichloromethane and a saturated solution of Panso ₃ . The aqueous phase was extracted with dichloromethane and the combined organic extracts were dried over MgSO ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUTC / hexanes 50-100%) using three columns simultaneously. The fractions containing impurities were collected and purified by flash chromatography using the aforementioned purification conditions. The fractions containing pure product were combined and concentrated to give the desired product as an orange solid (9.6 g, 85%).

¹ H NMR (400 MHz, COCl ₃ ) δ ppm: 8.90-8.84 (t, 1H), 8.55 (ά, 1 = 4.5 Hz, 1H), 8.29 (δ, 1H). 7.88-7.83 (t, 1H), 7.63 (δ, 1H), 7.58-7.53 (t, 2H), 7.35-7.31 (t, 1H), 7.23 (ά, 1 = 8.6 Hz, 2H), 6.89 (, 1 = 8.6 Hz, 2H), 6.19 (Lg. Δ, 1H), 5.82 (δ, 1H), 4.99 (δ, 2H), 3.90 (, 1 = 10.0 Hz, 2H), 3.80 (δ, 3H), 2.93-2.90 (t, 1H), 2.40 (δ, 3H), 2.362.28 (t, 1H), 2.22-2.20 (t, 2H), 1.89-1.88 (t, 1H), 1.80 (, 1 = 8.7 Hz, 2H), 1.38 (δ, 3H), 0.88–0.85 (t, 2H), 0.68–0.64 (t, 2H);

MS-IER: [M + H] + 619.3; calculated for [ΟΑΝΑ + Η] * 619.3.

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Obtaining typical compounds according to the invention

A1. N-Cyclopropyl-4- (7 - ((cyclopropylmethyl) amino) -5 - (((13.2K-2-hydroxycyclohexyl) amino) pyrazolo [1,5-a] pyrimidin-3-yl) -2-methylbenzamide hydrochloride

A mixture of tert-butyl (3-bromo-5 - (((13.2K) -2-hydroxycyclohexyl) amino) pyrazolo [1,5-a] pyrimidine-7yl) (cyclopropylmethyl) carbamate (8.48 g, 17.7 mmol), N-cyclopropyl-2-methyl-4- (4,4,5,5-tetramethyl1,3,2-dioxaboran-2-yl) benzamide (6.92 g, 23.0 mmol), RbCl ₂ brrHDHM (1.44 g, 1.76 mmol) and 2 M K ₃ PO ₄ (26.6 ml, 106.2 mmol) in THF (60 ml) were divided into four portions, each of which was placed in a sealed tube for microwave ovens. Each tube was filled with argon and heated in a microwave for 3 hours at 125 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The resulting crude product was purified by flash chromatography (gradient: EUTC / hexanes 20-100%) using two columns simultaneously, with a yellow solid being obtained.

The compound obtained in the previous step was dissolved in DCM (20 ml) and treated with TFA (20 ml) for 1 hour at room temperature. After completion of the reaction, the solvent was removed under vacuum and the crude product was re-dissolved in DCM (20 ml), neutralized by the addition of a saturated aqueous solution of sodium bicarbonate and extracted with ethyl acetate. The combined organic extracts were dried over HAZO ₄ , filtered and concentrated. This crude product was purified by flash chromatography (gradient: 50-100% EUAc / hexanes, then MeOH / DCM 010%) and triturated with HbO to give the title compound as a free base (white solid). The resulting free base was dissolved in a mixture of DCM (25 ml) and MeOH (50 ml), then HC1 (1 M E ( ₂ O, 2 eq.)) Was slowly added. The solvent was removed under vacuum to give the title compound in the form of the HC1-salt , in the form of a pale yellow solid (2.09 g, yield after 2 stages: 25%).

¹ H NMR (400 MHz, CO ₃ OO) δ ppm: 8.21 (δ, 1H), 7.51-7.37 (t, 3H), 5.67 (Lg. Δ, 1H), 4.04-3.82 (t, 2H) , 3.46-3.37 (t, 2H), 2.93-2.82 (t, 1H), 2.47 (δ, 3H), 1.89-1.64 (t, 6H), 1.61-1.42 (t, 2H), 1.31-1.21 (t, 1H), 0.88-0.78 (t, 2H), 0.70-0.58 (t, 4H), 0.46-0.36 (t, 2H);

MS-IER: [M + H] + 475.3; calculated for [С ₂₇ Н ₃₄ НЫ ₆ О ₂ + Н] + 475.3.

Purity according to HPLC: 98% at 254 nm.

A2. N-Cyclopropyl-4- (7 - ((cyclopropylmethyl) amino) -5- (pyridin-3-yloxy) pyrazolo [1,5a] pyrimidin-3-yl) -2-methylbenzamide hydrochloride

A mixture of tert-butyl (3-bromo-5- (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidine-7yl) (cyclopropylmethyl) carbamate (367 mg, 0.80 mmol), L-cyclopropyl-2- methyl 4- (4,4,5,5-tetramethyl1,3,2-dioxaboran-2-yl) benzamide (289 mg, 0.96 mmol), RbCl ₂ brrGDHM (65 mg, 0.080 mmol) and 2 M K ₃ PO ₄ (1.4 ml, 2.8 mmol) in THF (4 ml) were placed in a tightly closed microwave tube. The tube was filled with argon and heated in a microwave for 4 hours at 130 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. This crude product was purified by flash chromatography (gradient: EUTC / hexanes 20-100%) to give a yellow oil. The compound obtained in the previous step was dissolved in DCM (6 ml) and treated with TFA (2 ml) for 1 hour at room temperature. After completion of the reaction, the solvent was removed under vacuum and the crude product was re-dissolved in MeOH (4 ml), filtered and purified by preparative HPLC. Fractions were passed through a HornRac column and triturated with HbO to afford the title compound as a free base (white solid, 119 mg). Za- 19 029372

The portion of the resulting free base (67 mg, 0.15 mmol) was dissolved in MeOH (10 ml) and then HC1 (1 M E1 ₂ O, 2 eq.) was slowly added. This mixture was concentrated and triturated with E1 ₂ O to afford the title compound as the HC1 salt, as a yellow solid (55 mg, yield after 2 steps: 25%).

¹ H NMR (400 MHz, SOzOO) δ ppm: 9.10 (b, 1H), 8.82 (b, 1 = 6.0 Hz, 1H), 8.73-8.66 (t, 1H), 8.41 (b, 1H), 8.24-8.15 (t, 1H), 7.69 (b, 1H), 7.60 (b, 1 = 8.3 Hz, 1H), 7.23 (b, 1 = 8.0 Hz, 1n), 6.07 (b, 1H), 3.39 (b , 1 = 7.0 Hz, 2H), 2.89-2.78 (t, 1H), 2.29 (b, 3H), 1.37-1.23 (t, 1H), 0.86-0.75 (t, 2H), 0.70-0.56 (t, 4H ), 0.45-0.38 (t, 2H);

MS-IER: [M + H] + 455.2; calculated for [С26Н26Н ₆ О2 + Н] ⁺ 455.2.

Purity according to HPLC: 95.9% at 254 nm.

A3. 4- (5- (Cyclopentylamino) -7 - ((((1B, 3B) -3-hydroxy-3-methylcyclobutyl) methyl) amino) pyrazolo [1,5-a] pyrimidin-3-yl) -Y-cyclopropyl -2-methylbenzamide hydrochloride

A mixture of tert-butyl (3-bromo-5- (cyclopentylamino) pyrazolo [1,5-a] pyrimidin-7-yl) (((1B, 3B) -3 - ((tertbutoxycarbonyl) oxy) -3-methylcyclobutyl) methyl ) carbamate (0.40 g, 0.67 mmol), N-cyclopropyl-2methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxabolan-2-yl) benzamide (0.26 g, 0.88 mmol), Rbsl ₂ brrG-DHM (0.055 g, 0.067 mmol) and 2 M K ₃ PO ₄ (1 ml, 2.01 mmol) in THF (4 ml) were purged with argon and heated in a microwave for 3 h at 130 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried for ₂ Points _4, filtered and concentrated. This crude product was purified by flash chromatography (gradient: EUT / hex 20-100%) to give a yellow oil. The compound obtained in the previous step was dissolved in DCM (10 ml) and treated with TFA (3 ml) for 3 hours at room temperature. After completion of the reaction, the solvent was removed under vacuum and the resulting crude product was dissolved in MeOH (5 ml). This mixture was filtered and purified by preparative HPLC. The compound was passed through a HornRac cartridge and triturated with E1 ₂ O to give the title compound as a free base (white solid). The resulting free base was dissolved in MeOH (5 ml) and then HC1 (1 M E1 ₂ O, 2 eq.) Was slowly added. The solvent was removed in vacuo to afford the title compound as the HC1 salt, as a pale orange solid (75 mg, yield after 2 steps: 21%).

¹ H NMR (400 MHz, SOEOO) δ ppm: 8.18 (b, 1H), 7.42 (b, 3H), 5.52 (lg. B, 1H), 4.23–4.11 (t, 1H), 3.683.54 (t, 2H), 2.93-2.81 (t, 1H), 2.46 (b, 3H), 2.38-2.27 (t, 1H), 2.26-2.07 (t, 4H), 1.99-1.89 (t, 2H), 1.881 .59 (t, 6H), 1.35 (b, 3H), 0.86-0.79 (t, 2H), 0.66-0.58 (t, 2H);

MS-IER: [M + H] + 489.4; calculated for ^ 28 ^ 6 ^ 2 + 4 + 489.3.

Purity according to HPLC: 99% at 254 nm.

A4. N-Cyclopropyl-4- (7 - ((((1B, 3B) -3-hydroxy-3-methylcyclobutyl) methyl) amino) -5- (pyridine-3yloxy) pyrazolo [1,5-a] pyrimidine-3- Il) -2-methylbenzamide hydrochloride and its free base

A) Through the removal of the Vos-protective group.

A mixture of tert-butyl (3-bromo-5- (pyridin-3-yloxy) pyrazolo [1,5-a] pyrimidin-7-yl) (((1b, 3 B) -3 - ((tert-butoxycarbonyl) oxy) - 3-methylcyclobutyl) methyl) carbamate (0.23 g, 0.38 mmol), H-cyclopropyl-2methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl) Benzamide (0.15 g, 0.49 mmol), Rbsl ₂ brrG-DHM (0.15 g, 0.49 mmol) and 2 M K3PO4 (0.57 ml, 1.14 mmol) in THF (4 ml) purged with argon and heated in a microwave for 3 hours at 130 ° C. Water and ECUA were added, the phases were separated and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were dried over NAZO ₄ , filtered and concentrated. This crude product was purified by flash chromatography (gradient: EUTC / hexanes 20-60%) to give a yellow oil. The intermediate obtained in the previous step was dissolved in DCM (10 ml) and treated with TFA (3 ml) for 3 hours at room temperature.

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temperature After completion of the reaction, the solvent was removed under vacuum and the resulting crude product was dissolved in MeOH (5 ml). This mixture was filtered and purified by preparative HPLC. The compound was passed through a HornRac cartridge and triturated with E! ₂ About to obtain specified in the title compound in the form of a free base (white solid). The resulting free base was dissolved in MeOH (5 ml), and then HC1 (1 M E! ₂ O, 2 eq.) Was slowly added. The solvent was removed under vacuum to afford the title compound as the HC1 salt, as a beige solid (96 mg, yield after 2 steps: 47%).

¹ H NMR (400 MHz, SEZOE) δ ppm: 9.14 (Lg. Δ, 1Η), 8.89-8.82 (t, 1Η), 8.79-8.71 (t, 1), 8.40 (δ, 1Η), 8.31 -8.21 (t, 1Η), 7.68 (δ, 1Η), 7.59 (d, 1 = 9.5 Hz, 1Η), 7.23 (d, 1 = 8.0 Hz, 1Η), 6.06 (δ, 1), 3.56 (d, 1 = 6.5 Hz, 2H), 2.88-2.79 (t, 1Η), 2.40-2.31 (t, 1Η), 2.29 (δ, 3H), 2.26-2.18 (t, 2Η), 1.99-1.89 (t, 2Η) , 1.37 (δ, 3H), 0.85-0.76 (t, 2Η), 0.63-0.53 (t, 2Η);

MS-IER: [M + H] + 499.3; calculated for [C ₂₈ H _{3 (} L ₆ O ₃ + H] + 499.2.

Purity according to HPLC: 99.5% at 254 nm.

B) Through the removal of the RMV-protective group.

The mixture of No. cyclopropyl-4- (7 - ((((^ ^) - 3-hydroxy-3-methylcyclobutyl) methyl) (4-methoxybenzyl) amino) -5- (pyridin-3-yloxy) pyrazolo [1,5- a] pyrimidin-3-yl) -2-methylbenzamide (9.6 g, 15.5 mmol), TFA (50 ml) in ECE (1,2-dichloroethane) (70 ml) was heated in an oil bath for 4 h at 50 ° C. After completion of the reaction, the solvent was removed under vacuum and the resulting crude product was dissolved in a mixture of MeOH / DCM (100 ml / 25 ml). Then 2 M No. ₂ CO ₃ (150 ml) was added and the mixture was stirred for 30 minutes at room temperature. Then the reaction mixture was diluted with dichloromethane and the phases were separated. The aqueous phase was extracted with dichloromethane and the combined organic extracts were washed with water, dried over Mg ₈ O ₄ , filtered and concentrated. This crude product was triturated in a mixture of DCM / E! ₂ O (10 ml / 70 ml) and sonicated to give the title compound as a free base, as a yellowish solid (5.9 g, 77%).

¹ H NMR (400 MHz, CO ₃ OP) δ ppm: 8.58-8.53 (t, 1Η), 8.50-8.46 (t, 1Η), 8.36 (δ, 1Η), 7.86-7.80 (t, 1Η) , 7.76-7.72 (t, 1Η), 7.61-7.55 (t, 2Η), 7.18 (d, 1 = 8.0 Hz, 1Η), 5.92 (δ, 1Η), 3.52 (d, 1 = 6.8 Hz, 2H), 2.862.77 (t, 1Η), 2.38-2.28 (t, 1Η), 2.25 (δ, 3H), 2.24-2.18 (t, 2Η), 1.99-1.88 (t, 2Η), 1.37 (δ, 3H), 0.84-0.75 (t, 2Η), 0.64-0.54 (t, 2Η);

MS-IER: [M + H] + 499.2; calculated for [C ₂ 8NLO ₃ + H] + 499.2.

Purity according to HPLC: 96.1% at 235 nm.

The following compounds according to the invention were synthesized analogously to the compounds described in Examples A1-4_

Structure Example number MS calculated; MS-IER [M + H] ⁺ Salt form; HPLC purity II NMR ^but / hl ν m Y / Υ ^ΝΗ V ° A5 [ε ₂₈ Η ₃₀ Ν ₆ ο ₃ + Η] ⁺ 499.2; 499.3; free base; 98.9% at 254 nm (400 MHz, FCC) δ ppm: 8.55 (cM, 7 = 2.8, 0.4 Hz, 1H), 8.47 (άά, E = 4.8, 1.6 Hz, 1H), 8.33 (§, 1 H), 7.83-7.80 (w, 1H), 7.72 (δ, 1H), 7.58-7.55 (t, 2H), 7.18 (ά, E = 8.0 Hz, 1H), 5.89 (δ, 1H), 3.49 (ά, E = 7.6 Hz, 2H), 2.87-2.78 (t,

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2H), 2.28-2.23 (w, 5H), 1.97-1.92 (w,

2H), 1.36 (s, ZN), 0.82-0.77 (w, 2H),

0.62-0.58 (sh, 2H)

..θ ’

A6

[<7 ₂₇ Η ₂₈ Ν ₆ Ο ₃ + Η] ⁺

485.2;

485.3;

HCl salt;

98.8% at 254 nm

(400 MHz, ΓΩ ₃ ΟΩ) δ ppm: 9.13 (b, 7 = 2.3 Hz, 1H), 8.86 (b, = 5.8 Hz, 1H),

8.77-8.69 (sh, 1H), 8.40 (s, 1H), 8.26 (bb, = 8.8, 5.8 Hz, 1H), 7.67 (s, 1H), 7.59 (b, 8.0 Hz, 1H), 7.24 (b, 7 = 8.0

Hz, 1H), 6.07 (s, 1H), 4.53-4.39 (sh, 1H), 3.57 (b, 7 = 8.0 Hz, 2H), 2.89-2.79

A8

[<7 ₂₇ Η ₃₄ Ν ₆ Ο ₂ + Η] ⁺

475.3;

475.4;

NSGsol;

99.3% at 254 nm

A11

[<7 ₂₇ Η ₂₈ Ν ₆ Ο ₃ + H] ⁺ 485.2 485.2;

NSGsol;

98.6% at 254 nm

(sh, 1H), 2.76-2.61 (sh, 1H), 2.30 (s, ZN), 2.29-2.21 (sh, 2H), 2.21-2.08 (sh , 2H), 0.85-0.78 (w, 2H), 0.65-0.58 (w, 2H)

(400 MHz, SODO) δ ppm: 8.22 (s, 1H), 7.51-7.36 (w, 3N), 5.68 (s, 1H), 3.77 (s, 2H ), 3.37-3.33 (sh, 2H), 2.93-2.84 (sh, 1H), 2.47 (s, 3N), 2.07-1.95 (sh, 4H), 1.93-1.72 (sh, 4H), 1.34-1.19 (sh, 1H), 0.88-0.78 (sh, 2H), 0.69-0.59 (sh, 4H ), 0.45-0.37 (w, 2H)

(400 MHz, SODO) δ ppm: 9.14 (s, 1H), 8.85 (b, 7 = 5.5 Hz, 1H), 8.79-8.72 (sh, 1H), 8.40 (h, 1H), 8.30-8.21 (sh, 1H), 7.68 (h, 1H), 7.59 (b, 7 = 7.8 Hz, 1H), 7.23 (b, 7 = 8.0 Hz, 1H), 6.06 (s, 1H), 4.17-4.06 (sh, 1H), 3.54 (b, 7 = 6.5 Hz, 2H) , 2.88-2.78 (sh, 1H), 2.53-2.43 (sh, 2H), 2.29 (s, 4H), 1.81-1.68 (sh, 2H), 0 , 84-0.77 (sh, 2H), 0.62-0.56 (sh, 2H)

ΝΗ

A12

[С ₂₇ Н ₃₄ И ₆ О ₂ + Н] ⁺

475.3;

475.4;

NSGsol;

97.5% at 254 nm

A13

[<7 ₂₈ Η ₃₆ Ν ₆ Ο ₂ + H] ⁺ 489.3;

489.4

TFA salt;

98.2% at 254 nm

(400 MHz, CO ₃ (W) δ ppm: 8.18 (s, 1H), 7.55-7.29 (w, 3N), 5.54 (LP, W, 1H), 4, 50-4.37 (t, 1H), 4.26-4.07 (t, 1H), 3.62 (b, 7 = 6.8 Hz, 2H), 2.92-2.83 (t, 1H), 2.75-2.61 (t, 1H), 2.46 (s, ZN), 2.31-2.02 (t, 6H), 1.92-1.56 (t, 6H) , 0.91-0.75 (t, 2H), 0.71-0.52 (t, 2H) (400 MHz, CO ₃ (W) δ ppm: 8.18 (W, 1H), 7.48-7.36 (t, ЗН), 5.52 (t, 1H), 4.234.10 (t, 1H), 3.65-3.57 (t, 2H), 2.932.75 (t, 2H), 2.46 (s, ZN), 2.31-2.21 (t, 2H), 2.20-2.11 (t, ZN), 2.06-1.89 (t, 2H) , 1.89-1.50 (t, 6H), 1.37 (z.ZN), 0.87-0.79 (t, 2H), 0.68-0.56 (t, 2H)

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Example B. Analysis of TTC inhibition.

The active TTC was obtained from 1puPodpodsp as a recombinant protein of the unprocessed human TTC with C8T (glutathione-g-transferase) at the amino terminus. Human TTC (residues 1-275) labeled with IMO (a small ubiquitin-like modifier) containing 6 histidine residues at the amino terminus were expressed in E. soy and purified to homogeneity> 95% using No. ² + agarose, gel filtration and ion exchange chromatography.

TTC activity was measured using indirect EB1AA (enzyme-linked immunosorbent assay) as a registration system. C8T-TTK (0.68 nM) were incubated in the presence of 16 μM ATP (81 cat. No. A7699) or 100 μM ATP, 50 mM Nerek, pH 7.2, 1 mM ESTA, 10 mM MdCl ₂ and 0.1% Pluronic in a 96-well microtiter plate pre-coated with human TTC (residues 1-275) labeled with I.IMO containing 6 histidine residues at the amino terminus.

The reaction was carried out for 30 minutes, then the tablet was washed 5 times with washing buffer (phosphate-buffered saline supplemented with 0.2% Tween 20) and incubated for 30 minutes with a primary antibody (Ce11 δί§ηα1ίη§ cat. No. 9381) in dilution of 1: 3000. The tablet was washed 5 times with washing buffer, incubated for 30 minutes in the presence of a secondary antibody conjugated with horseradish peroxidase (“cat. No. 1721019, concentration 1: 3000), washed 5 more times with washing buffer and incubated in the presence of TMB (3, 3 ', 5,5'-tetramethylbenzidine) (81dsha cat. No. T0440), as a substrate. A colorimetric reaction was carried out for 5 minutes, then a stop solution (0.5N. Sulfuric acid) was added and the signal was quantitatively recorded at 450 nm using a monochromatic plate reader or plate reader with a filter (Motioner Iueyuek M5 or WeightTech ETH880, respectively).

The inhibitory ability of the compounds was determined either at a fixed concentration (10 μM) or at various inhibitor concentrations (usually by dose-response titration at 10 points in the concentration range from 0.5 to 0.001 μM). The compounds were preincubated in the presence of the enzyme for 5 minutes, then ATP was added and the remaining activity was quantified using the activity assay described above. % inhibition of the compound was determined using the following formula:% inhibition = 100 x (1 - (experimental value - background value) / (high active control - background value)). The value of IC ₅₀ (the concentration required for 50% inhibition) was determined using the fit to the nonlinear 4-parametric logistic curve (ХЬй14, ГОВ§) using the formula: (А + (В / (1 + ((х / С) ^Л И)) ))), where A = background value, B = range, C = inflection point, And = curve fitting parameter.

The table below (see below) lists the IR ₅₀ values of typical compounds obtained using 100 μM ATP. The ranges of the IC ₅₀ values are indicated as “A” if the IC ₅₀ values are less than or equal to 0.1 μM, as “B”, if the IC ₅₀ values are greater than 0.1 μM and less than or equal to 0.5 μM, and as “C” if IC50 values are greater than 0.5 μM. The ranges of IC50 values marked with an asterisk were obtained using 16 μM ATP for analysis (§1§ta cat. No. А7699).

Example B. Data for typical compounds according to the invention, obtained using a cancer cell line.

Breast cancer cells (MIA-MB-468), colon cancer cells (HCT116) and ovarian cancer cells (OUSAK-3) were seeded (from 1000 to 4000 cells in a volume of 80 μl / well, depending on the growth rate of the cells) in 96-well plate and incubated for 24 hours, then an aliquot of the compound was applied on top. Compounds, in the form of 10 mM stock solutions in 100% DMSO, were diluted with cell growth medium IMEM (modified by Dulbecco's method Eagle's medium) (Pigodip, WigPd.Y., Canada) containing 10% PBV (fetal calf serum), to concentrations in range from 50 nM to 250 μM. Aliquots (20 μl) from each dilution were applied on top of the cells, pre-seeded in a volume of 80 μl in 96-well plates, to final compound concentrations in the range from 10 nM to 50 μM. Cells were cultured for 5 days and then the analysis was performed using the sulforhodamine B dye (QCB) to determine the inhibitory activity of the compounds with respect to cell growth.

Sulforhodamine B (obtained from §1 dta, Oakchee,, Canada) is a water-soluble dye that binds to the basic amino acids of cellular proteins. Accordingly, the colorimetric measurement of the bound dye allows to determine the total mass of proteins and, therefore, the number of cells. Cells were fixed with ίη 8 Yi by slowly pumping out the culture medium and adding 50 μl of 10% trichloroacetic acid (THU) cooled to 0 ° C per well and incubated for 30-60 min at 4 ° C. The plates were washed five times with water and left to air dry for 5 minutes. 50 μl of a 0.4% (w / v) GC solution in 1% (v / v) acetic acid was added to each well and incubated for 30 minutes at room temperature to complete the staining reaction. After staining, the plates were washed four times with 1% acetic acid to remove unbound dye and then left to air dry for 5 minutes. The bound dye was transferred to the solution by adding 100 μl of 10 mM Tris pH 10.5 per well. Optical density was measured at 570 nm.

- 23 029372

The percentage (%) of the relative growth inhibition was calculated by comparing with samples containing only DMSO-treated cells (100%). For compounds with cytotoxic activity, values of ΟΙ _{50 were} determined. Values of ΟΙ _{50 were} calculated using the StarBab program ΡΚΙ8Μ (StarBab 8 Dayt, 1 ps., San Diego, California, USA). ΟΙ ₅₀ (growth inhibition) —concentration of the compound that causes 50% inhibition of cell growth.

The table (see below) shows the ranges of values of ΟΙ ₅₀ compounds described in the examples obtained in the analysis of breast cancer cell lines (ΜΌΑ-ΜΒ-468), colon cancer cell lines (HCT116) and ovarian cancer cell lines (OUSAC). 3). The data show that the studied compounds differ in levels of growth inhibition / cytotoxic activity with respect to breast cancer cells, colon cancer and ovarian cancer. Ranges of values ΟΙ ₅ ο indicated as "A" if CE is less than the value ₀ or equal to 0.1 M, as "B" if the value is greater than 0.1 CP ₀ pM and less than or equal to 0.5 M, and as the "C , "if ΟΙ _{50 is} greater than 0.5 μM.

Example G. Data on the inhibition of typical cell compounds that initiate the development of a tumor, colon cancer and ovary

Substances and techniques.

Treated flasks of T-75 and 96-well plates for non-tissue and tissue cultures were obtained from UUK Vitamin A supplement B-27, MEM ΝΕΑΑ, sodium pyruvate, B-glutamine, N2 supplement, penicillin-streptomycin, and Fungisone / amphotericin B were obtained from ΙηνίΙΐΌβοη. A mixture of lipids, heparin and ECP was obtained from §1§ta; URNS was received from ΒΌ Vu8aepse8. Cells initiating the development of a tumor (TU) from the colon in accordance with standard methods were maintained using treated T-75 flasks for non-tissue cultures in OMEM: P12 medium containing 0.2X B-27 supplement, 4 μg / ml heparin, 1X MEM 1, 1X sodium pyruvate, 1 mM glutamine, 10 pg / μl CPR, 20 pg / μl ECP, supplement 1X N2, a mixture of lipids, penicillin-streptomycin and fungizone / amphotericin B. TY from the ovary in accordance with standard methods was maintained using processed flasks T-75 for tissue cultures in the environment EMEM: P12. containing 1X B-27 supplement, 4 μg / ml heparin, 20 pg / μl LRSP, 20 pg / μl ECP and penicillin-streptomycin.

Protocol analysis.

The compounds described herein were dissolved in DMSO and then diluted with cell culture media to determine their CP ₀ . Colon TY was trypsinized and seeded into 4,000 cells / well treated non-woven 96-well plates for non-tissue cultures. After 24 h, the compound was added to the cell culture in different concentrations and the final concentration of EM8O was adjusted to 0.1%. Then the cells were cultured at 37 ° C for 9 days. TY from the ovary was trypsinized and seeded into treated 96-well tissue culture plates at 1000 cells / well. After 24 h, the compound was added to the cell culture in different concentrations and the final concentration of EM8O was adjusted to 0.1%. Then the cells were cultured at 37 ° C for 6 days. Cell viability was assessed using the alamar blue indicator: 10 µl alamar blue was added to each well. After incubation for 4 h at 37 ° C, fluorescence was recorded using a wavelength of 544 nm for excitation and measuring the emission at a wavelength of 590 nm. SCO (growth inhibition) was calculated using the program SgarRab Rp8t 4.0. Data on the inhibition of cell growth compounds described in this document are shown in the table below.

The table (see below) shows the ranges of CP _{0 of the} compounds described in the examples obtained in the analysis of TY (Coop 12 and Oyap 2393Α). Ranges of values ₀ CP indicated as "A" if the value of CP is less than ₀ or equal to 0.1 M, as "B" if the value is greater than 0.1 CP ₀ pM and less than or equal to 0.5 M, and as the "C "if the CP ₀ values are greater than 0.5 µM.

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! η νΐίΓΟ activity of the compounds described in the examples

Example Range 1С50 for TTK Line Range cancer cells Sbp range for cells initiating tumor development MOA-MB468 HCT116 OUSAK-W Ον ap 2393A Cop 12 A1 BUT BUT BUT BUT BUT BUT A2 BUT BUT BUT BUT ΝΏ ΝΏ AZ BUT BUT BUT BUT ΝΌ ΝΌ A4 BUT BUT BUT BUT BUT Νϋ A5 BUT BUT BUT AT ΝΏ ΝΏ A6 BUT BUT BUT BUT BUT BUT A8 BUT AT AT WITH ΝΏ Νϋ A11 BUT BUT BUT BUT ΝΏ Νϋ A12 BUT BUT BUT BUT ΝΩ Νϋ A13 BUT AT BUT AT ΝΏ Νϋ

N0 - not defined.

Claims (9)

CLAIM 1. A compound represented by the following structural formula: or its pharmaceutically acceptable salt, where K ¹ represents -ΝΗ-CH3-Su; Su represents C ₃ -C ₄ -cycloalkyl, optionally having as substituents one or two groups selected from (C ₁ -C ₃ ) alkyl and hydroxyl; K ² is -O-pyridinyl, ^ H- (C ₂ -C ₆ ) hydroxyalkyl, optionally having cyclopropyl or isopropyl as a substituent, or ^ H- (C ₃ -C ₆ ) cycloalkyl, optionally having hydroxy as a substituent or (C ₁ -C ₂ ) hydroxy alkyl; K ^{4 is} selected from a hydrogen atom, a halogen atom, and (C 1 -C 3) alkyl; K ^a is cyclopropyl. 2. The compound according to claim 1, in which K ⁴ represents a chlorine atom or methyl. 3. A compound represented by the following structural formula: or its pharmaceutically acceptable salt. 4. A compound represented by the following structural formula: - 25 029372 or its pharmaceutically acceptable salt. 5. A compound represented by the following structural formula: or its pharmaceutically acceptable salt. 6. The compound represented by the following structural formula: or its pharmaceutically acceptable salt. 7. Pharmaceutical composition for treating cancer, containing a compound according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier or diluent. 8. A method of treating cancer, comprising administering to a subject in need an effective amount of a compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7. 9. The method of claim 8, wherein the cancer is pancreatic cancer, prostate cancer, lung cancer, melanoma, breast cancer, colon cancer or ovarian cancer.