EA036435B1 - Heterocyclic amides useful as protein modulators - Google Patents

Abstract

The invention relates to a compound which is (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide, having the structure stated below, or a pharmaceutically acceptable salt thereof. Furthermore, the invention relates to a pharmaceutical composition comprising the claimed compound and the use of the claimed compound for treating a STING-mediated disease.

Description

Related applications

This application claims the priority of U.S. Provisional Application No. 62/319358, filed April 7, 2016, U.S. Provisional Application No. 62/461301, filed February 21, 2017, and U.S. Provisional Application No. 62/461975, filed February 22, 2017. which are incorporated herein by reference in their entirety.

The technical field to which the invention relates

The present invention relates to heterocyclic amides that are useful as modulators of transmembrane protein 173 (TMEM173), which is also known as STING (interferon gene stimulator), and methods for their preparation and use.

State of the art

Vertebrates are constantly under the threat of microorganisms entering their body, and they have developed immune defense mechanisms to eliminate infectious pathogens. In mammals, this immune system consists of two links: innate immunity and adaptive immunity. The innate immune system is the first line of defense that is initiated by pattern recognition receptors (PRRs), which detect ligands from pathogens as well as damage-associated molecular structures (Takeuchi O. et al., Cell, 2010: 140, 805-820). A growing number of these receptors have been identified, including Toll-like receptors (TLRs), C-type lectin receptors, retinoic acid-induced gene-1 (RIG-I) receptors, and NOD-like receptors (NLRs), and double-stranded DNA sensors. PRR activation results in up-regulation of genes involved in the inflammatory response, including genes for type 1 interferons, pro-inflammatory cytokines, and chemokines that suppress the replication of pathogens and promote adaptive immunity.

The STING adapter protein (stimulator of interferon genes), also known as TMEM 173, MPYS, MlTA and ERIS, has been identified as a central signaling molecule in mediating the response of the innate immune system to cytosolic nucleic acids (Ishikawa H. and Barber GN, Nature, 2008 : 455, 674-678; WO2013 / 1666000). STING activation leads to upregulation of the IRF3 and NFkB pathways, which, in turn, leads to the induction of interferon-β and other cytokines. STING is key to providing responses to the cytosolic DNA of the pathogen or host, and unusual nucleic acids called cyclic dinucleotides (CDNs).

CDNs were initially identified as bacterial secondary messengers responsible for the regulation of multiple responses in the prokaryotic cell. Bacterial CDNs, such as c-di-GMP, are symmetrical molecules characterized by two 3'-5'phosphodiester bonds.

Direct activation of STING by bacterial CDNs has recently been confirmed by X-ray crystallography (Burdette D.L. and Vance R.E., Nature Immunology, 2013: 14, 19-26). Thus, bacterial CDNs and their analogs have attracted interest as potential vaccine adjuvants (Libanova R. et al., Microbial Biotechnology 2012: 5, 168-176; WO2007 / 054279; WO2005 / 087238).

More recently, the response to cytosolic DNA has been extensively investigated and has been shown to involve the generation, by an enzyme called cyclic GMP-AMP synthetase (cGAS, previously known as C6orfl50 or MB21D1), a new mammalian CDN signaling molecule identified as cGAMP. which then activates STING. Unlike bacterial CDNs, cGAMP is an unsymmetrical molecule characterized by mixed 2 ', 5' and 3 ', 5' phosphodiester bonds (Gao P. et al., Cell, 2013: 153, 1094-1107). The interaction of cGAMP (II) with STING has also been shown by X-ray crystallography (Cai X. et al., Molecular Cell, 2014: 54, 289296).

Interferon was first described as a compound that can protect cells from viral infection (Isaacs & Lindemann, J. Virus Interference, Proc. R. Soc. Lon., S. Biol. Sci. 1957: 147, 258267). In humans, type I interferons are a family of related proteins encoded by genes on chromosome 9 and which encode at least 13 isoforms of interferon alpha (IFNa) and one isoform of interferon beta (IFNe). Recombinant IFNa was the first officially approved

- 1 036435 as a biological therapeutic agent and provided an important therapy for viral infections and cancer. As well as direct antiviral activity on cells, interferons are known to be effective modulators of the immune response, acting on cells of the immune system.

Administration of a small molecule compound that can modulate the innate immune response, including the activation or inhibition of the production of type I interferon and other cytokines, can be an important strategy for the treatment or prevention of human diseases, including viral infections and autoimmune diseases. This type of immunomodulatory strategy has the potential to identify compounds that may be useful not only for innate immunity in infectious diseases, but also in cancer (Zitvogel L., et al., Nature Immunology, 2015, 15 (7), p405414), allergic diseases (Moisan J. et al., Am. J. Physiol. Lung Cell Mol. Physiol., 2006: 290, L987995), neurodegenerative diseases such as amyotrophic lateral sclerosis and multiple sclerosis (Lemos H. et al., J Immunol., 2014: 192 (12), 5571-8; Cirulli E. et al., Science, 2015: 347 (6229), 1436-41, Freischmidt A., et al., Nat. Neurosci., 18 ( 5), 631-6), other inflammatory conditions such as irritable bowel syndrome (Rakoff-Nahoum S., Cell., 2004, 23, 118 (2): 229-41) and as vaccine adjuvants (Persing et al. Trends Microbiol. 2002: 10 (10 Suppl), S32-7 and Dubensky et al., Therapeutic Advances in Vaccines, published online September 5, 2013).

STING is essential for host defense against microbes, including protection against DNA and RNA viruses and bacteria (see Barber et al., Nat. Rev. Immunol., 2015: 15 (2): 87-103, Ma and Damania, Cell Host & Microbe, 2016: 19 (2) 150-158). Viruses belonging to the families Herpesviridae, Flaviviridae, Coronaviridae, Papillomaviridae, Adenoviridae, Hepadnaviridae, Ortho- and Paramyxoviridae, and Rhabdoviridae have developed mechanisms of inhibition of STING-mediated production of interferon type I by the host and al. 2016: 7: 10680; Ma et al., PNAS 2015: 112 (31) E4306-E4315, Wu et al, Cell Host Microbe 2015: 18 (3) 333-44, Liu et al, J. Virol. 2016: 90 (20) 9406-19, Chen et al., Protein Cell, 2014: 5 (5) 369-81, Lau et al., Science, 2013: 350 (6260) 568-71; Ding et al., J. Hepatol., 2013: 59 (1) 52-8; Nitta et al., Hepatology, 2013 57 (1) 46-58, Sun et al., PloS One, 2012: 7 (2) e30802, Aguirre et al., PloS Pathog, 2012: 8 (10) e1002934; Ishikawa et al., Nature, 2009: 461 (7265) 788-92). Thus, activation of STING by a small molecule may be useful in the treatment of these infectious diseases.

In contrast, increased and prolonged production of type I IFN is associated with a variety of chronic infections, including infections caused by mycobacteria (Collins et al., Cell Host Microbe, 2015: 17 (6) 820-8); Wassermann et al. Cell Host Microbe 2015: 17 (6) 799-810; Watson et al., Cell Host Microbe, 2015: 17 (6) 811-9), Franciselles (Storek et al., J. Immunol., 2015: 194 (7) 3236-45; Jin et al., J. Immunol. ., 2011: 187 (5) 2595-601), chlamydia (Prantner et al., J. Immunol., 2010: 184 (5) 2551-60; plasmodia (Sharma et al., Immunity, 2011: 35 (2) 194-207 and HIV (Herzner et al., Nat. Immunol., 2015 16 (10) 1025-33, Gao et al., Science, 2013: 341 (6148) 903-6. Similarly, excess production of type I interferon occurs in patients with complex forms of autoimmune disease.

The results of genetic studies in humans and experiments in animal models support the hypothesis that inhibition of STING leads to a decrease in the production of type I interferon, which, in turn, controls autoimmune diseases (Crow YJ et al., Nat. Genet., 2006; 38 (8) 38917-920, Stetson DB et al., Cell, 2008; 134, 587-598). Therefore, STING inhibitors provide treatment for patients with chronic production of type I interferon and pro-inflammatory cytokines associated with infections or complex autoimmune diseases.

Allergic diseases are associated with an impaired Th2 immune response to allergens. Th2 responses are associated with increased levels of IgE, which, through its effect on mast cells, increase hypersensitivity to allergens, leading to the development of symptoms such as those seen in allergic rhinitis and asthma. In healthy individuals, the immune response to allergens is more balanced with a mixed Th2 / Th1 response and a regulatory T cell response. It was shown that induction of type I interferons leads to a decrease in the production of Th2-type cytokines in the local environment and promotes the induction of Thl / Treg responses. In this context, induction of type I interferons, for example, activation of STING, can be beneficial in the treatment of allergic diseases such as asthma and allergic rhinitis (Huber J.P. et al., J. Immunol., 2010: 185, 813-817).

Compounds that bind to STING and function as an agonist have been shown to induce type I interferons and other cytokines when incubated with human PBMCs. Compounds that induce human interferons can be useful in the treatment of various disorders, for example, in the treatment of allergic diseases and other inflammatory conditions, for example, allergic rhinitis and asthma, in the treatment of infectious diseases, neurodegenerative diseases, precancerous diseases and cancer, and may also be useful. as an immunogenic composition or vaccine adjuvants. Compounds that bind to STING can function as antagonists and can be useful in the treatment of, for example, autoimmune diseases. It is envisaged that targeting STING with activating or inhibitory agents could be a promising approach to treat diseases and conditions in which modulation of the type I IFN pathway is effective, including inflammatory, allergic and autoimmune diseases, infectious diseases, cancer, precancerous diseases, and as an immunogenic composition or vaccine adjuvants.

With skin cancer and various viral infections of the skin, a special immune environment is created, and the activation of a local immune response to lesions may be an actual therapeutic approach. STING agonists can be used to treat viral warts, superficial malignant skin diseases, and precancerous actinic keratoses. Through a dual mechanism of action, STING activation (eg, by delivery with a microneedle patch or topical drug) can be used to control HPV directly through the production of Type I interferon with its antiviral activity and indirectly by enhancing the adaptive immune response below the activation of the innate immune system. The STING agonist can activate the innate immune response at the lesion and regulate the T-cell response against HPV.

Recent data indicate that spontaneous activation of the STING pathway in tumor resident dendritic cells leads to the production of type I IFNs and adaptive immune responses against tumors. In addition, activation of this pathway in antigen presenting cells (APCs) in the tumor microenvironment leads to subsequent priming of T cells for tumor associated antigens (Corrales and Gajewsli, Clin. Cancer Res., 21 (21); 4774-9, 2015).

International patent applications WO 02 014/093936, WO 02014/189805, WO2013 / 185052, US2014 / 0341976, WO 2015/077354, PCT / EP2015 / 062281 and GB 1501462.4 disclose certain cyclic dinucleotides and their use in inducing an immune response by activating STING ...

The compounds of the present invention modulate STING activity and, accordingly, can have a useful therapeutic effect in the treatment of diseases, disorders and / or conditions in which modulation of STING (interferon gene stimulator) is useful, for example, for the treatment of inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous diseases, and as vaccine adjuvants.

The essence of the invention

The invention relates to a compound that is (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3methyl-1 H-pyrazole-5 -carboxamido) -7- (3-morpholinopropoxy) -1H- benzo [d] imidazol-1-yl) but-2-en-1 yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -7-methoxy-1 H-benzo [ d] imidazole-5-carboxamide having the structure:

or a pharmaceutically acceptable salt thereof.

It should be understood that references herein to a compound of the invention and a pharmaceutically acceptable salt thereof include the compound of the invention as the free base or as salts thereof, for example, as a pharmaceutically acceptable salt thereof. Thus, in one embodiment, the invention relates to a compound of the invention in the form of a free base. In yet another embodiment, the invention relates to a compound of the invention and a pharmaceutically acceptable salt thereof.

A compound of the invention, or a pharmaceutically acceptable salt thereof, are STING modulators. Therefore, the present invention relates to a compound of formula (I-N), (I-P) or (I), or a salt thereof, in particular a pharmaceutically acceptable salt thereof, for use in therapy. This invention particularly relates to the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, as an active therapeutic compound in the treatment of a disease or disorder mediated by STING, in particular for use in the treatment of a disease mediated by an agonist or antagonistic effect of STING. The invention also relates to a compound of formula (IN), (IP) or (I) or a salt thereof, in particular a pharmaceutically acceptable salt, for use in the manufacture of a medicament for the treatment of a disease or disorder mediated by STING.

The invention also relates to a method of treating a STING-mediated disease, comprising administering a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, to a person in need thereof. Such diseases or disorders mediated by STING include cancer and solid tumor.

The present invention also relates to a pharmaceutical composition for the treatment of a STING-mediated disease comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

Detailed description of the invention

According to one aspect of the present invention, the present invention relates to a compound which is (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- ( 3morpholinopropoxy) -1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -7-methoxy-1H-benzoU] imidazole-5-carboxamide having the structure:

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a pharmaceutically acceptable salt (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy ) -1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1 H-benzo [ d] imidazole-5-carboxamide.

In another embodiment, the compound is (E) -1- (4- (5-carbamoyl2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) -1H- benzo (^] imidazol-1yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzoU] imidazole-5carboxamide.

According to another aspect of the present invention, the present invention relates to a pharmaceutical composition for the treatment of a STING-mediated disease, comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

According to another aspect of the present invention, the present invention relates to a method of treating a STING-mediated disease, comprising administering a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, to a person in need thereof.

In one embodiment, the disease is cancer.

In yet another embodiment, the disease is a solid tumor.

According to another aspect of the present invention, the present invention relates to the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the treatment of a STING-mediated disease.

According to another aspect of the present invention, the present invention relates to the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a STING-mediated disease.

In one embodiment, the disease is a solid tumor.

In yet another embodiment, the disease is cancer.

The term pharmaceutically acceptable refers to those compounds, substances, compositions and dosage forms that, based on the results of careful medical evaluation, are suitable for use in contact with tissues of humans and animals without excessive toxicity, irritation or other problems or complications, commensurate with a reasonable benefit / risk ratio. ...

It should be understood that references herein to a compound of the invention and a pharmaceutically acceptable salt thereof include the compound of the invention as the free base or as salts thereof, for example, as a pharmaceutically acceptable salt thereof. Thus, in one embodiment, the invention relates to a compound of the invention in the form of a free base. In yet another embodiment, the invention relates to a compound of the invention and a pharmaceutically acceptable salt thereof.

Consequently, the present invention includes a compound of the invention, as defined herein, in any salt or non-salt form and in any physical form and mixture of various forms. Although included in the present invention, it should be understood that a compound of iso-4 036435 as defined herein, in any salt or non-salt form and in any physical form, can have different levels of activity, different bioavailability and different handling properties. for formulation purposes.

Suitable pharmaceutically acceptable salts of a compound of the invention may include acid addition salts or base addition salts. For reviews of suitable pharmaceutically acceptable salts see Berge et al., J. Pharm. Sci., 66: 1-19, (1977) and monographs by P.H. Stahl and C.G. Wermuth, Eds., Handbook of Pharmaceutical Salts: Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim / Zurich: Wiley-VCH / VHCA (2002).

Salts of a compound of the invention containing a basic amino group or other basic functional group can be prepared by any suitable method known in the art, such as treatment of the free base with a suitable inorganic or organic acid. Examples of such pharmaceutically acceptable salts include acetate, adipate, ascorbate, aspartate, benzenesulfonate, benzoate, camphor, camphorsulfonate (camsylate), caprate (decanoate), caproate (hexanoate), caprylate (octanoate), carbonate, bicarbonate, cinnamate, citrate, cyclate dodecyl sulfate (estolate), ethane-1,2-disulfonate (edisylate), ethanesulfonate (esylate), formate, fumarate (hemifumarate, etc.), galactarate (mucat), gentisate (2,5-dihydroxybenzoate), glucoheptonate (glucept ), gluconate, glucuronate, glutamate, glutarate, glycerophosphate, glycolate, hippurate, hydrobromide, hydrochloride (dihydrochloride, etc.), hydroiodide, isobutyrate, lactate, lactobionate, laurate, maleate, mandelonate, malonate, malonate , naphthalene-1,5-disulfonate (naphizylate), naphthalenesulfonate (napsylate), nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, phosphate (diphosphate, etc.), propionate, pyroglutamate, salicitate, sebacate, stearate, succinate , sulfate, tartrate, thiocyanate, p-toluenesulfonate (tosylate), undecylenate, 1-gi Droxy 2-naphthoate, 2,2-dichloroacetate, 2-hydroxyethanesulfonate (isethionate), 2-oxoglutarate, 4-acetamidobenzoate and 4-aminosalicylate.

The invention includes within its scope all possible stoichiometric and non-stoichiometric salt forms (eg, hydrobromide, dihydrobromide, fumarate, hemifumarate, etc.) of the compound of the invention.

In general, a pharmaceutically acceptable salt can be readily prepared using a suitable acid or base, whichever is more appropriate. The resulting salt can be crystallized or precipitated from solution, or obtained by trituration, and can be isolated by filtration or evaporation of the solvent.

Since the compounds of the present invention are intended for use in pharmaceutical compositions, it is easy to understand that each of them should preferably be obtained in substantially pure form, for example, at least 60% pure, more suitably at least 75% pure. and preferably at least 85%, even more preferably at least 98% purity (% by weight based on total weight). The impure formulations of the compounds can be used to obtain purer forms for use in pharmaceutical compositions.

As used herein, the terms cancer, neoplasm, and tumor are used interchangeably and, in singular or plural form, refer to cells that have undergone a malignant transformation that renders them pathological to the host. Primary cancer cells can be easily distinguished from non-cancerous cells using well known techniques, especially histological examination. The definition of a cancer cell, as used herein, includes not only a primary cancer cell, but any cell derived from a precursor of a cancer cell. The definition includes metastatic cancer cells and in vitro cultures and cell lines derived from cancer cells. With respect to the type of cancer that usually manifests itself as a solid tumor, the clinically detectable tumor is a tumor that is found on the basis of the tumor mass; for example, by procedures such as computed tomography (CT), magnetic resonance imaging (MRI), x-rays, ultrasound, or palpation on physical examination, and / or which can be detected by the expression of one or more tumor-specific antigens in the sample, taken from the patient. Tumors can be hematopoietic (or hematologic or blood-related) tumors, for example, malignant tumors derived from blood cells or immune cells, which can be categorized as liquid tumors. Specific examples of clinical conditions based on hematologic tumors include leukemias such as chronic myelocytic leukemia, acute myelocytic leukemia, chronic lymphocytic leukemia, and acute lymphocytic leukemia; plasma cell cancers such as multiple myeloma, MGUS, and Waldenstrom's macroglobulinemia; lymphomas such as non-Hodgkin's lymphoma, Hodgkin's lymphoma, and the like.

The cancer can be any type of cancer in which there is an abnormal number of blast cells or unwanted cell proliferation, or is diagnosed as hematologic cancer, including both lymphoid and myeloid cancers. Myeloid neoplasms include, but are not limited to, acute myeloid (or myelocytic, or myelogenous, or myelogenous) leukemia (undifferentiated or differentiated), acute promyeloid (or promyelocytic or promyelogenous or promyeloblastic) leukemia, acute myelogenous

- 5 036435 (or myelomonoblastic) leukemia, acute monocytic (or monoblastic) leukemia, erythroleukemia and megakaryocytic (or megakariblastic) leukemia. Collectively, these leukemias can refer to acute myeloid (or myelocytic or myelogenous) leukemia (AML). Myeloid neoplasms also include myeloproliferative disorders (MPD), which include, but are not limited to, chronic myelogenous (or myeloid) leukemia (CML), chronic myelomonocytic leukemia (CMML), essential thrombocythemia (or thrombocytosis) and polycythemia vera (polycythemia vera). Myeloid neoplasms also include myelodysplasia (or myelodysplastic syndrome or MDS), which may be referred to as refractory anemia (RA), refractory anemia with excess blasts (RAEB) and refractory anemia with excess blasts with transformation (RAEBT); and myelofibrosis (MFS) with or without agogenic myeloid metaplasia.

Hematopoietic malignancies also include lymphoid neoplasms, which can affect the lymph nodes, spleen, bone marrow, peripheral blood, and / or extranodal regions. Lymphoid neoplasms include B-cell neoplasms, which include, but are not limited to, B-cell non-Hodgkin's lymphomas (B-NHL). B-NHLs can be indolent (or low grade), intermediate grade (or aggressive), or high grade (very aggressive).

Indolent B-cell lymphomas include follicular lymphoma (FL); small lymphocytic lymphoma (SLL); marginal zone lymphoma (MZL), including nodal MZL, extranodal MZL, splenic MZL and splenic MZL with villous lymphocytes; lymphoplasmic lymphoma (LPL); and lymphomas with mucosal-associated lymphoid tissue (MALT or marginal extranodal lymphoma). Intermediate grade B-NHLs include mantle cell lymphoma (MCL) with or without leukemia, diffuse large cell lymphoma (DLBCL), follicular large cell lymphoma (stage 3 or 3B), and primary mediastinal lymphoma (PML). High-grade B-NHLs include Burkitt's lymphoma (BL), Burkitt's lymphoma like lymphoma, intersected nucleus small cell lymphoma (SNCCL), and lymphoblastic lymphoma. Other B-NHLs include immunoblastic lymphoma (or immunocytoma), primary effusion lymphoma, HIV-associated (or AIDS-related) lymphomas, and post-transplant lymphoproliferative disease (PTLD) or lymphoma. B-cell neoplasms also include, but are not limited to, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), Waldenstrom macroglobulinemia (WM), hairy cell leukemia (HCL), large granular lymphocyte leukemia (LGL), or acute lymphocytic or lymphoblastic) leukemia and Castelmann's disease. NHL may also include T-cell non-Hodgkin lymphomas (T-NHL), which include, but are not limited to, T-cell non-Hodgkin's lymphoma (NOS), peripheral T-cell lymphoma (PTCL), anaplastic large cell lymphoma (ALCL), angioimmunoblastic lymphoid disorder (AILD), NK / T-cell lymphoma of the nasal type, gamma / delta lymphoma, cutaneous T-cell lymphoma, fungal mycoses and Sesari's syndrome.

Hematopoietic malignancies also include Hodgkin's lymphoma (or disease), including classic Hodgkin's lymphoma, sclerosing nodular Hodgkin's lymphoma, mixed cell Hodgkin's lymphoma, Hodgkin's lymphocytic lymphoma (LP), Hodgkin's lymphoma and lymphocyte depletion Hodgkin's lymphoma. Hematopoietic malignancies also include diseases or tumors associated with abnormal proliferation of plasma cells, such as multiple myeloma (MM), including smoldering MM, monoclonal gammopathy of undetermined (or unknown or unclear) nature (MGUS), plasmacytoma (bone, extramedullary), lymphoplasmic lymphoma (LPL), Waldenstrom's macroglobulinemia, plasma cell leukemia, and primary amyloidosis (AL). Hematopoietic malignancies can also include other tumors containing additional hematopoietic cells, including polymorphonuclear leukocytes (or neutrophils), basophils, eosinophils, dendritic cells, platelets, red blood cells, and natural killer cells. Tissues that include hematopoietic cells, referred to herein as hematopoietic cell tissues, include bone marrow; peripheral blood; thymus; and peripheral lymphoid tissues such as the spleen, lymph nodes, lymphoid tissues associated with the mucosa (such as gut-associated lymphoid tissues), tonsils, Peyer's patches, and lymphoid tissues associated with other mucous membranes, eg, bronchial lining.

Examples of cancers and conditions in which the compounds of the present invention can have potentially beneficial anticancer effects include, but are not limited to, cancers of the lung, bone, pancreas, skin, head, neck, uterus, ovary, stomach, colon, breast. gland, esophagus, small intestine, digestive tract, endocrine system, thyroid gland, parathyroid gland, adrenal gland, urethra, prostate, penis, testicle, ureters, bladder, kidney, or liver; rectal cancer; anal cancer; carcinoma of the fallopian tubes, endometrium, cervix, vagina, vulva, renal pelvis, renal cell carcinoma; sarcoma of soft tissues; myxome;

- 6 036435 rhabdomyoma; fibroma; lipoma; teratoma; cholangiocarcinoma; hepatoblastoma; angiosarcoma; hemangioma; hepatoma; fibrosarcoma; chondrosarcoma; myeloma; chronic or acute leukemia; lymphocytic lymphomas; primary CNS lymphoma; neoplasms of the central nervous system; spinal cord tumors; squamous cell carcinomas; synovial sarcoma; malignant pleural mesothelioma; brain stem glycous; pituitary adenoma; bronchial adenoma; chondromatous hamartoma; mesothelioma; Hodgkin's disease, or a combination of one or more of the above cancers.

In one aspect, the person has a solid tumor. In one aspect, the tumor is selected from head and neck cancers, gastric cancer, melanoma, renal cell carcinoma (RCC), esophageal cancer, non-small cell lung cancer, prostate cancer, colorectal cancer, ovarian cancer, and pancreatic cancer. In one aspect, the human has one or more of the following diseases: colorectal cancer (CRC), esophageal, cervical, bladder, breast cancer, head and neck cancers, ovarian cancer, melanoma, renal cell carcinoma (RCC), squamous cell EC, non-small cell lung carcinoma, mesothelioma and prostate cancer. In yet another aspect, the human has a fluid tumor such as diffuse large B cell lymphoma (DLBCL), multiple myeloma, chronic lymphoblastic leukemia (CLL), follicular lymphoma, acute myeloid leukemia, and chronic myeloid leukemia.

A therapeutically effective amount means an amount of a compound that, when administered to a patient in need of such treatment, is sufficient for effective treatment or prophylaxis as defined herein. Thus, for example, a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, is the amount of an agent of the invention that, when administered to a human in need thereof, is sufficient to modulate STING activity such that the disease mediated by this activity is suppressed, alleviated. or prevented. The amount of a particular compound that will correspond to such an amount will vary depending on such factors as the particular compound (e.g., efficiency (pIC _50), effective (EC50) and the biological half-life of the specific compound), disease condition and its severity, the identity ( for example, age, size and body weight) of a patient in need of treatment, but can nevertheless be determined by a person skilled in the art. Similarly, the duration of treatment and the period of administration (the period between doses and the time of administration of doses, for example, before / s / after meals) of the compound will vary depending on the identity of the mammal in need of treatment (for example, body weight), the particular compound and its properties ( for example, pharmacokinetic properties), disease or disorder and its severity, as well as the specific composition and method that can be used, but can nevertheless be determined by the person skilled in the art.

Treating or treating means at least ameliorating the disease or disorder in the patient. Methods of treatment for ameliorating a disease or disorder include the use of compounds of the present invention in any conventional manner, for example, to slow, treat, or treat a disease or disorder mediated by STING, as described above. In one embodiment, treating, treating, or treating cancer refers to ameliorating cancer, eliminating or ameliorating one or more symptoms of cancer, slowing or eliminating the progression of cancer, and delaying the relapse of the condition in a patient or subject who has previously had or is diagnosed with cancer.

The compounds of the invention can be administered by any suitable route of administration, including both systemic and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or inhalation, and is usually given by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the lungs of a patient, whether inhalation through the mouth or through the nasal passages. Topical administration includes application to the skin.

In addition to the above-described modes of administration suitable for the treatment of cancer, the pharmaceutical compositions can be adapted for administration by intratumoral or peritumoral injection. It is contemplated that intratumoral or peritumoral injection of a compound of the present invention directly into or adjacent to one solid tumor can elicit an immune response that can attack and destroy tumor cells throughout the body, significantly reducing and in some cases eliminating the tumor from the diseased subject. Activating the immune system to eliminate tumors at a distant site is widely recognized as an abscopal effect and has been demonstrated in animals with multiple therapeutic modalities (van der Jeught et al., Oncotarget, 2015, 6 (3), 1359-1381). Another advantage of local or intratumoral or peritumoral administration is the ability to achieve equivalent efficacy at significantly lower doses, which minimizes or eliminates the adverse events that can occur at much higher systemic doses (Marabelle, A., et al., Clinical Cancer Research, 2014, 20 (7), p 1747-1756).

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The compounds of the invention can be administered once or in accordance with a dosing regimen in which a number of doses are administered at different intervals over a period of time. For example, doses can be administered one, two, three, or four times a day. Doses can be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of the particular compound, such as absorption, distribution and half-life, which can be determined by one of ordinary skill in the art. In addition, suitable dosage regimens, including the duration of such regimens, are administered since the compound of the invention depends on the disease or disorder being treated, the severity of the disease or disorder being treated, the age and physical condition of the patient being treated, the patient's medical history, to be treated, the nature of the concomitant therapy, the desired therapeutic effect, and similar factors, which are within the knowledge and experience of one skilled in the art. It will also be understood by those skilled in the art that suitable dosage regimens may need to be adjusted based on the response of an individual patient to the dosing regimen or over time as each individual patient needs to change the treatment regimen. The total daily dose is 1 to 2000 mg, preferably the total daily dose is 1 to 250 mg.

For use in therapy, the compounds of the invention will usually, but not necessarily, be formulated in a pharmaceutical composition for administration to a patient. Therefore, the invention also relates to pharmaceutical compositions comprising a compound of the invention and at least one pharmaceutically acceptable excipient.

Pharmaceutical compositions of the invention can be prepared and packaged in bulk form, where an effective amount of a compound of the invention can be recovered and then administered to a patient, for example in the form of powders, syrups, and injection solutions. Alternatively, pharmaceutical compositions of the invention can be prepared and packaged in a unit dosage form. For oral administration, for example, one or more tablets or capsules can be administered. A dosage of a pharmaceutical composition contains at least a therapeutically effective amount of a compound of the present invention (i.e., a compound of the invention, or a pharmaceutically acceptable salt thereof). When formulated in a unit dosage form, the pharmaceutical compositions may contain from 1 mg to 1000 mg of a compound of the present invention.

As provided herein, unit dosage forms (pharmaceutical compositions) containing from 1 to 1000 mg of a compound of the invention can be administered one, two, three or four times a day, preferably one, two or three times a day, and more preferably one or twice daily, to treat a disease or disorder mediated by STING.

Pharmaceutical compositions of the invention typically contain one compound of the invention. However, in some embodiments, the pharmaceutical compositions of the invention comprise more than one compound of the invention. In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional therapeutic agents (eg, pharmaceutically active compounds).

As used herein, the term pharmaceutically acceptable excipient refers to a pharmaceutically acceptable material, composition, or carrier used in imparting shape or consistency to a pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when mixed in such a way as to avoid interactions that could significantly reduce the efficacy of a compound of the invention when administered to a patient and interactions that result in pharmaceutical compositions that are not pharmaceutically acceptable. In addition, each excipient must, of course, be of a sufficiently high purity to be pharmaceutically acceptable.

The compounds of the invention and the pharmaceutically acceptable excipient or excipients are usually formulated in a dosage form adapted for administration to a patient by the desired route of administration. Typical dosage forms include those that are adapted for (1) oral administration such as tablets, capsules, capsules, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and starches; (2) parenteral administration, such as sterile solutions, suspensions and powders for reconstitution; (3) transdermal administration, such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation administration such as aerosols and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

Suitable pharmaceutically acceptable excipients will vary depending on the particular dosage form chosen. In addition, suitable pharmaceutically acceptable excipients can be selected for the particular function they can perform in the composition. For example, some pharmaceutically acceptable excipients can be selected for their ability to facilitate the production of uniform dosage forms. Some pharmaceutically

- 8 036435 Acceptable excipients can be selected based on their ability to facilitate the preparation of stable dosage forms. Certain pharmaceutically acceptable excipients can be selected based on their ability to facilitate transfer or transport of a compound or compounds of the invention after administration to a patient from one organ or body part to another organ or body part. Certain pharmaceutically acceptable excipients can be selected for their ability to improve patient compliance.

Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavors, flavoring agents , coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, viscosity-increasing agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents. One of ordinary skill in the art will understand that some pharmaceutically acceptable excipients may serve more than one function and may serve alternative functions depending on how much excipient is present in the formulation and what other ingredients are present in the formulation.

Those skilled in the art have the knowledge and skill in the art to enable them to select suitable pharmaceutically acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of sources that are available to the person skilled in the art that describe pharmaceutically acceptable excipients, and these can be useful in the selection of suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared using techniques and methods known to those of skill in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one aspect, the invention relates to a solid oral dosage form, such as a tablet or capsule, containing an effective amount of a compound of the invention and a diluent or excipient. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (eg, corn starch, potato starch, and pregelatinized starch), cellulose and its derivatives (eg, microcrystalline cellulose), calcium sulfate and dibasic calcium phosphate. The solid dosage form for oral administration may additionally contain a binder. Suitable binders include starch (eg, corn starch, potato starch and pregelatinized starch), gelatin, gum arabic, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (eg, microcrystalline cellulose). The solid dosage form for oral administration may additionally contain a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmellose, alginic acid, and sodium carboxymethyl cellulose. The solid dosage form for oral administration may additionally contain a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate and talc. For example, tablets can be prepared by conventional methods and formulated as follows: compound, 5 mg; microcrystalline cellulose, 100 mg; lactose, 100 mg; sodium starch glycolate, 30 mg; magnesium stearate, 2 mg; total weight: 237 mg. Capsules can be prepared by conventional means and are formulated as follows: compound, 15 mg; dried starch, 178 mg; magnesium stearate, 2 mg; total weight: 195 mg.

Certain compounds of the invention may be effective immunomodulators and, accordingly, care should be taken when handling them.

Examples of

The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled person in the preparation and use of the compounds, compositions and methods of the present invention. While specific embodiments of the present invention have been described, one skilled in the art realizes that various changes and modifications can be made without departing from the spirit and scope of the invention.

It should be understood that some of the compounds of the invention may be effective immunomodulators and therefore should be handled with care.

The reactions described herein are useful for preparing compounds of the invention having several different substituents (eg, R ¹ , R ² , etc.) as defined herein. One skilled in the art will understand that if a particular substituent is incompatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is resistant to the reaction conditions. Suitable protecting groups and methods for protecting and deprotecting various substituents using such suitable protecting groups are well known to those skilled in the art, examples of which can be found in T. W. Greene Protective

Groups in Organic Synthesis (4th edition, J. Wiley and Sons, 2006). Unless otherwise stated, all starting materials were obtained from commercial suppliers and used without further purification.

Certain intermediates described herein form another aspect of the invention.

General synthetic methods

Compounds of the present invention can be prepared using the synthetic procedures shown below in the reaction schemes, which can be easily adapted to obtain other compounds of the invention, based on the knowledge of specialists in the field of organic chemistry. The syntheses outlined in these schemes are useful for preparing compounds of the invention having numerous different R groups using suitable precursors, which are suitably protected, if necessary, to ensure compatibility with the reactions described herein. Subsequent deprotection, when necessary, provides compounds with the properties disclosed herein. Although the schemes are shown with compounds of formula (I-N), (I-P) or (I) only, they illustrate methods that can be used to prepare compounds of the invention. Intermediates (compounds used in the preparation of compounds of the invention) can also be in the form of salts.

Method 1: The corresponding nitrohalobenzamide (1A) can be treated with an amine (exemplary allylamine) under alkaline or metal-mediated coupling conditions to provide aniline (1B). Subsequent reduction of the nitro group under appropriate conditions gives dianiline (1C). Interaction with cyanogen bromide provides aminobenzimidazole (1D). The peptide combination of aminobenzimidazole and pyrazolecarboxylic acid (IE) forms the amidobenzimidazole monomer (1F). The cross-metathesis reaction between two molecules (1F) gives an unsaturated dimer (1G), which can be hydrogenated to give a saturated 1H dimer.

Method 1:

1H

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Method 2: Two molecules of the corresponding nitrohalobenzamide (2A) can be treated with a diamine under alkaline conditions or under metal-mediated coupling conditions to give biscoupled aniline (2B). The subsequent bis-reduction of nitro groups under appropriate conditions can provide dianiline (2C). Interaction with cyanogen bromide gives aminobenzimidazole dimer (2D). A peptide combination of a bis-amine (2D) and two pyrazole carboxylic acid molecules (2E) gives an amidobenzimidazole dimer (2F).

Method 2:

Method 3: fluoronitrobenzamide (3A) is treated with monoprotected diamine under alkaline or metal-mediated coupling conditions to produce aniline (3B). The subsequent reduction of the nitro group under appropriate conditions can provide dianiline (3C). Interaction with cyanogen bromide gives aminobenzimidazole (3D). Peptide coupling of amine (3D) and pyrazolecarboxylic acid (3E) gives amidobenzimidazole (3I), which, under appropriate conditions, can be deprotected by removal of the amino protecting group to give amine (3G). The amine (3G) can be combined with the appropriate halogenated nitrobenzamide (3H) to provide compound (31); reduction of the nitro group can give dianiline (3J). Treatment of compound (3J) with cyanogen bromide leads to the formation of aminobenzimidazole (3K), which can be treated with pyrazolecarboxylic acid (3E) under amide coupling conditions to give an unsymmetrical dimer (3L).

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Method 3:

Method 4: The corresponding nitrohalobenzamide (4A) is treated with a monoprotected diamine under alkaline or metal-mediated coupling conditions to give allylaniline (4B). The subsequent reduction of the nitro group under appropriate conditions provides dianiline (4C).

Interaction with cyanogen bromide gives aminobenzimidazole (4D). Peptide coupling of amine (4D) and pyrazolecarboxylic acid (4E) gives amidobenzimidazole (4F), which can be deprotected by removal of the amino protecting group under appropriate conditions to give amine (4G). The amine (4G) can be combined with fluoronitrobenzamide (4H) to give compound (41), and then reduction of the nitro group gives dianiline (4J). Treatment of (4J) with cyanogen bromide affords aminobenzimidazole (4K), which can be treated with pyrazolecarboxylic acid (4E) under amide coupling conditions to give an unsymmetrical dimer (4L).

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Method 4:

Method 5: Two molecules of the corresponding phenol (5A) are reacted with a biselectrophile such as dibromopropane to give an ester-linked dimer (5B). The dimer (5B) is then reacted with the corresponding diamine to give the macrocycle (5C). Reduction of the nitro group followed by treatment with cyanogen bromide gives bis-aminobenzimidazole (5D). Amide coupling with a suitable acid (5E) gives macrocyclic bis-amidobenzimidazole (5F).

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Method 5:

Method 6: Bis-pyrazolecarboxylic acid 6A (Method 8) is reacted with aminobenzimidazole dimer (6B) under amide coupling conditions to give macrocyclic amidobenzimidazole (6C), where each R can be the same or different.

Method 6:

Method 7: bis-pyrazolecarboxylic acid 7A (method 8) is reacted with two molecules of aminobenzimidazole (7B) under amide coupling conditions to obtain a pyrazole-linked dimer (7C). The ring closure metathesis reaction produces an unsaturated macrocycle (7D) that can be hydrogenated to give a saturated macrocyclic amidobenzimidazole (7E).

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Method 7:

Method 8: Substituted (pent-4-yn-1-yl) -1H-pyrazole carboxylate (8C) can be prepared by Nalkylating substituted 1H-pyrazole carboxylate (8A) using (5-chloropent-1-yn-1yl) trimethylsilane followed by desilylation. 4-Iodo-1H-pyrazolecarboxylate (8F) can be prepared by esterification of the corresponding 1H-pyrazolecarboxylic acid (8D) followed by iodination using 1-iodopyrrolidine-2,5-dione.

Palladium catalyzed coupling of substituted (pent-4-yn-1-yl) -1H-pyrazole carboxylate (8C) with 4-iodo-1H-pyrazole carboxylate (8F) gives alkynyl-linked bis-pyrazole (8G). Hydrogenation of the alkynyl-linked bis-pyrazole followed by hydrolysis provides the bis-pyrazole carboxylic acid used in methods 6 and 7 (6A / 7A) above.

Method 8:

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Method 9: The corresponding nitrohalobenzamide (9A) can be treated with a monoprotected diamine (such as 9B) under alkaline conditions or under metal-mediated coupling conditions to produce a combined aniline (9C). Subsequent deprotection of the primary amine provides amine 9D. The second nitrohalobenzamide (9E) can be reacted with amine 9D under alkaline conditions or under metal-mediated coupling conditions to provide the bis-nitrodimeric benzamide (9F). Dianiline (9G) can provide double reduction of nitro groups under appropriate conditions. Interaction with cyanogen bromide gives aminobenzimidazole dimer (9H). The peptide combination of bis-amine 9H and two molecules of pyrazolecarboxylic acid (9I) forms an amidobenzimidazole dimer (9J).

Method 9:

Method 10: Tetraaniline (10A, prepared by Method 9 or other general method) can be treated with isothiocyanate (10B) until the reaction is complete. After completion of step 1, EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) are added and the reaction mixture is stirred until complete to give the amidobenzimidazole dimer (10 ° C.).

Method 10:

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Method 11: All variables have the values defined in formula (I-N), (I-P) or formula (I). The appropriately substituted halogenated nitrobenzamide (11A) is reacted with a monoprotected diamine such as compound 11B to provide nitroaniline 11C. Deprotection by removal of the amino protecting group provides amine 11D, which can be reacted with a halogenated nitrophenyl derivative 11E to give bis-nitro-nF. Reduction of both nitro groups will give bis-aniline 11G, which is treated with cyanogen bromide to give bisaminobenzimidazole 11H. Amide coupling with a pyrazole carboxylic acid such as 11I gives the substituted amidobenzimidazole dimer 11J. When the corresponding functional groups are present in 11J, further functionalization of these groups makes it possible to obtain additional compounds such as 11K.

Method 11:

Method 12: In method 12, R ^C2 = R ¹⁴ , R ¹⁷ = R ¹⁵ and R ¹⁶ = R ^C1 , all other variables have the meanings defined in formula (IN), formula (IP) or formula (I). Tetraaniline (12A, prepared by Method 11, 16 or other general method) can be treated with an isothiocyanate such as 12V until the reaction is complete. After completion of Step 1, EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) are added and the reaction mixture is stirred until the reaction is complete to give the amidobenzimidazole dimer (12 ° C.).

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Method 12:

Method 13: In method 13, all variables have the meanings defined in formula (I-N), formula (I-P), or formula (I). Using this method, substituted macrocyclic compounds can be prepared. The substituted halogenated nitrophenyl derivative (13A) is treated with an appropriate diamine (13B) containing a linker group (B) between the two amino groups to provide amine 13C. Reduction of the nitro group followed by treatment with cyanogen bromide can give aminobenzimidazole 13D. Amide coupling of compound 13D and a mono-bis-pyrazole carboxylic acid (such as 13E) containing a linker group (C) between the two pyrazoles will provide amidobenzimidazole (13F). The deprotection of the amino group allows coupling to a second substituted halogennitrophenyl (13G) to give the nitroester 13H. Reduction of the nitro group of compound 13H followed by treatment with cyanogen bromide affords aminobenzimidazole 13I. Then hydrolysis of the pyrazole ester provides the formation of a macrocyclic amide to obtain a macrocyclic amidobenzimidazole 13J. When suitable functional groups are present at 13J, further functionalization of these groups allows additional compounds such as 13K to be prepared.

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Method 13:

R ¹⁴

Method 14: —O — M ¹ is defined as optionally substituted (C | —C ₆ alkyl) oxy as defined for R ^a1 when q is 0 in formula (I) of formula (IN) or formula (IP). All other variables have the meanings defined in formula (IN), formula (IP) or formula (I). A dimeric amidobenzimidazole containing phenol, such as compound 14A, prepared by one of the general synthetic methods described herein, can be alkylated in phenol using a suitable alkylating agent such as an alkyl bromide and a base such as potassium carbonate. When suitable functional groups are present in 14B, further functionalization of these groups allows for the preparation of additional compounds.

Method 14:

_R C1 _R C1

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Method 15: All variables have the meanings defined in formula (I-N), formula (I-P) or formula (I). An appropriately substituted halogenated nitrophenyl derivative (15A) is reacted with a diamine containing a linker group (B), such as 15B, to provide bis-nitrodimer 15C. Reduction of both nitro groups will give tetraaniline 15D, which can be converted to amidobenzimidazole dimer (15E) in one of two ways: 1) treatment with cyanogen bromide to give bis-aminobenzimidazole followed by amide coupling with a pyrazole carboxylic acid such as 15F; or 2) treatment with isothiocyanate (15G) until the reaction is complete, then adding EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) and the reaction mixture is stirred until the reaction is complete. When suitable functional groups are present in 15E, further functionalization of these groups allows for the preparation of additional compounds.

Method 15:

2.EDC, base

Method 16: All variables have the meanings defined in formula (I-N), formula (I-P) or formula (I). The appropriately substituted halogenated nitrophenyl derivative (16A) is reacted with a monoprotected diamine containing a linker group (B), such as 16B, to provide nitroaniline 16C. Removal of protection by removal of the amino protecting group provides amine 16D, which can be reacted with a halogenated nitrophenyl derivative 16E to provide bis-nitrodimer 16F. Reduction of both nitro groups will provide tetraaniline 16G, which can be converted to amidobenzimidazole dimer (16H) in one of two ways: 1) treatment with cyanogen bromide to give bis-aminobenzimidazole followed by amide coupling with a pyrazole carboxylic acid such as 16I; or 2) treatment with isothiocyanate (16J) until the reaction is complete, then adding EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) and the reaction mixture is stirred until the reaction is complete. When suitable functional groups are present in 16H, further functionalization of these groups allows for the preparation of additional compounds.

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Method 16:

Method 17: M ² is C1-C ₆ alkyl, or COOM ² can be any ester that is inactive for hydrogenolysis of the benzyl ester. All other variables have the meanings defined in formula (IN), formula (IP) or formula (I). Substituted pyrazole ester such as (pent-4-yn-1-yl) -1H-pyrazole carboxylate (17D) can be prepared by esterification of substituted 1H pyrazole carboxylate (17A) to give ester 17B followed by N-alkylation under the conditions Mitsunobu's reactions. 4-Iodopyrazole ester (17G) can be prepared by esterification of the corresponding pyrazole carboxylic acid (17E) followed by iodination using 1iodopyrrolidine-2,5-dione (NIS). Palladium catalyzed coupling of an alkylated pyrazole such as 17D with a 4-iodo-pyrazole ether (17G) gives the bound bis-pyrazole (17H). Reduction and hydrogenolysis of the bound bis-pyrazole will give bis-pyrazolecarboxylic monoacid (17I).

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Method 17:

Method 18: All variables have the values defined in formula (A). (E) -1- (4- (5-carbamoyl-2 (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole-1- yl) but-2en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1 H-benzo ^] imidazole-5-carboxamide (18A) can be treated with methanesulfonyl chloride and triethylamine. After completion of stage 1, the resulting mesylate (18B) is treated with amine (NHRARB) and K ₂ CO ₃ and the reaction mixture is heated at 5080 ° C until the reaction is complete to obtain the target compounds (18 ° C).

Method 18:

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Method 19: All variables have the meanings defined in formula (I-N), formula (I-P) or formula (I). A suitably substituted bis-aminobenzimidazole (19B) containing a linker group (B), prepared by one of the methods described herein, is reacted with bis-pyrazole (19A) containing a linker group (C) and amide coupling reagents to give macrocyclic bis-amidobenzimidazole.

Method 19:

R ⁵ pz

19A 19C

Method 20: All variables have the meanings defined in formula (I-N), formula (I-P) or formula (I). The appropriately substituted halogenated nitrophenyl derivative (20A) is reacted with a monoprotected diamine containing a linker group (B), such as 20B, to provide nitroaniline 20C. Reduction of the nitro group under appropriate conditions gives dianiline 20D, which can be converted to amidobenzimidazole 20F in one of two ways: 1) treatment with cyanogen bromide followed by amide coupling with a pyrazole carboxylic acid such as 20E; or 2) treatment with isothiocyanate (20L) until the reaction is complete, then adding EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) and stirring the reaction mixture until the reaction is complete. Removal of protection by removal of the amino protecting group provides amine 20G, which can be reacted with a halogenated nitrophenyl derivative 2H to give dimeric nitroaniline 20I. Reduction of the nitro group will provide bis-aniline 20J, which can be converted to an amidobenzimidazole dimer (20K) in one of two ways: 1) treatment with cyanogen bromide to give bis-aminobenzimidazole followed by amide coupling with a pyrazole carboxylic acid such as 20M; or 2) treatment with isothiocyanate (20N) until the reaction is complete, then adding EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) and the reaction mixture is stirred until the reaction is complete. When suitable functional groups are present at 20K, further functionalization of these groups makes it possible to obtain additional compounds.

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Method 20:

R ⁴

R ⁴

RA1 20A

R ³

1. BrCN

2. Amide

H-.N B

R ⁴

О ₂ 1

R ³

1. BrCN

2. Amide combination

Recovery of nitro group

2. EDC,

Method 21: All variables have the meanings defined in formula (I-N), formula (I-P) or formula (I). Two molecules of a suitable functionalized nitrohalophenyl (21A) are dimerized to provide bis-nitrodimer 21B containing a linker group (A). Then compound 21B is reacted with an amine or diamine to obtain dianilin 21C. Reduction of nitro groups gives tetraaniline 21C, which can be converted to amidobenzimidazole dimer (21G) in one of two ways: 1) treatment with cyanogen bromide to give bis-aminobenzimidazole followed by amide coupling with a pyrazole carboxylic acid such as 21E; or 2) treatment with isothiocyanate (21F) until the reaction is complete, then adding EDC (or other suitable coupling reagent) and triethylamine (or other suitable base) and the reaction mixture is stirred until the reaction is complete. When suitable functional groups are present on 21G, further functionalization of these groups allows additional compounds to be obtained. A common example of dimerization is the reaction of a suitable nitrophenol (21H) with a bis-halide and a base to give bis-phenol dimer 21J.

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Method 21:

2. Amide combination

2.edc, base

Method 22: All variables have the values defined in formula (I-N), formula (I-P), or formula (I). A suitably substituted aminobenzimidazole (22A), prepared by one of the methods described herein, is reacted with bis-pyrazole (22B) containing a linker group (C) and amide coupling reagents to form a dimeric bis-amidobenzimidazole. When suitable functional groups are present at 22C, further functionalization of these groups allows additional compounds to be prepared.

Method 22:

Method 23: All variables have the meanings as defined for formula (I-N), formula (I-P), or formula (I). A suitably substituted aminobenzimidazole (23A), prepared by one of the methods described herein, is reacted with bis-pyrazole monocarboxylic acid (23B) containing a linker group (C) and amide coupling reagents to provide an amidobenzimidazole ester such as 23C. Hydrolysis of the pyrazole ester gives the acid 23D, which can be combined with a second aminobenzimidazole (23E) to give the dimeric bisamidobenzimidazole (23F). When suitable functional groups are present at 23F, further functionalization of these groups allows for the preparation of additional compounds.

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Method 23:

Method 24: M ² is C ₁ -C ₆ alkyl, or COOM ² can be any ester that is inactive for hydrogenolysis of the benzyl ester. All other variables have the meanings defined for formula (IN), formula (IP), or formula (I). Substituted pyrazole ester such as (pent-4-yn-1-yl) -1H-pyrazole carboxylate (24D) can be prepared by esterification of substituted 1H-pyrazole carboxylate (24A) to give ester 24B followed by Nalkylation under appropriate conditions. such as the presence of an alkyl halide and a base. In the case of using (5-chloropent-1-yn-1-yl) trimethylsilane, subsequent desilylation gives the pyrazole ester 24D. The 4-iodopyrazole ester (24G) can be prepared by esterification of the corresponding pyrazole carboxylic acid (24E) followed by iodination using 1-iodopyrrolidine-2,5-dione (NIS). Palladium catalyzed coupling of an alkylated pyrazole such as 24D with a 4-iodopyrazole ether (24G) gives the bound bis-pyrazole (24H). Reduction and hydrogenolysis of the bound bispyrazole will provide bis-pyrazole-monocarboxylic acid (241), which can be further hydrolyzed to give bis-pyrazole-diacid 24 J.

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Method 24:

It will be understood that in any of the methods described above, the exact order of the synthetic steps by which the various groups and moieties are introduced into the molecule can be changed. A person skilled in the art should understand that subsequent transformations and reactions will not affect the groups or moieties introduced in one step of the process and, accordingly, the choice of the order of the synthetic steps.

Intermediate and final compound names described here were generated using ChemDraw Pro 12.0.2.1076 Plug-In inside Perkin Elmer E-Notebook or MarvinSketch 5.11.4_b82 (Chemaxon) chemical naming programs.

Those skilled in the art will understand that in some cases, these programs may refer to the structurally depicted compound as a tautomer or isomer of that compound. It should be understood that any reference to a named compound or structurally depicted compound is intended to encompass all tautomers or isomers of such compounds and any mixtures of tautomers and / or isomers thereof.

The definitions for LC-MS analysis conditions are listed below and apply to all compounds.

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LC-MS method LC-MS Method A Wavelength 214 nm and 254 nm Device Agilent 1200-6110 Column Halo C18 4.6x50 μm Flow rate 1.8 ml / min Gradient method Time (min) ACN (0.05% TPA) H ₂ 0 (0.05% TPA) 0 5 95 one 95 5 2 95 5 2.5 5 95 LC-MS method LC-MS Method B Wavelength 214 nm and 254 nm Device Shimadzu 2020 Column Halo C18 4.6x50 μm Flow rate 1.5 ml / min Gradient method Time (min) ACN (0.05% TPA) H ₂ 0 (0.05% TPA) 0 5 95 one 95 5 4 95 5 4.5 5 95 5 5 95

LC-MS method: LC-MS method C Instrument

LC: Shimadzu ΙΟΑνρ (controller, pumps and UV detector) UV: Shimadzu lOAVp (214 nm) ELS Sedere Sedex 75C (45C) MS: PE Sciex 150EX Single Quadrupole Polarity (positive); mode (profile); scanning time (0.33 s); stage (0.2 m / z) capillary B (5500); cone B (25-45) or Waters ZQ single quadrupole Polarity (positive); mode (continuum); scanning time (0.25 s); stage (0.2 m / z) capillary B (3500); cone B (25-35) Autosampler CTC Leap; loop 3 μl; standard injection volume = 2 μl (standard) Column: Thermo Hypersil Gold (С18, 20x2.1 mm, diameter particles 1.9 μm) Heater Phenomenex 50-55 ° C Solvent A: H ₂ O, 0.02% TPA Solvent B: MeCN, 0.02% TPA Gradient: Time (min) Flow (ml / min) Solvent B 0.02 1.4 4, 0 1.90 95, 0 1.91 4, 0 2.00 Stop

LC-MS Method: LC-MS Method D Instrument

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LC: Waters Acquity Binary Solvent Manager Column Manager 55C Autosampler CTC Leap PAL UV: Waters Acquity PDA (210-360 nm) ELS Waters Acquity ELSD (50C) or Sedere Sedex 75C (45C) MS: Waters Acquity SQD Polarity (positive or negative); mode (continuum); scan time (0.15 s); capillary B (3500); cone B (25-35); Column: Thermo Hypersil Gold (С18, 20x2.1 mm, diameter particles 1.9 μm) Solvent A: H2O, 0.02% TPA Solvent IN: MeCN, 0.02% TPA Gradient Time (min) Flow (ml / min) Solvent B 0.02 16 2.0 1.90 95, 0 1.91 Stop 4.0

LC-MS method: LC-MS method E

Device

LC: Waters Acquity I-Class Binary Solvent Manager Column Manager 55C Autosampler: CTC Leap PAL 3 UV: Waters Acquity PDA (210-360 nm) ELS Waters Acquity ELSD (50C) or Sedere Sedex 75C (45C) MS: mass detector Waters Acquity QDa Polarity (positive or negative); mode (continuum); scan time (10 Hz); capillary kV (0.8); cone B (12); Column: Thermo Hypersil Gold (C18, 20x2.1 mm, particle diameter 1.9 μm) Solvent A: H ₂ O, 0.02% TPA Solvent IN: MeCN, 0.02% TPA Gradient Time (min) Flow (ml / min) Solvent B 0.02 16 0.5 1.90 90 to 95 1.91 Stop 0.5

LC-MS method: LC-MS method F

Device

LC: Waters Acquity Binary Solvent Manager Column Manager 55C Autosampler: CTC Leap PAL 3 UV: Waters Acquity PDA (210-360 nm) ELS Waters Acquity ELSD (50C) or Sedere Sedex 75C (45C) MS: Waters Acquity SQD Polarity (positive or negative); mode (continuum); scan time (0.15 s); capillary B (3500); cone B (22-35); Column: Waters VEN (C18, 30x2.1 mm, particle diameter 1.7 μm) Solvent A: H ₂ O, 0.02% TPA Solvent B: MeCN, 0.02% TPA Gradient Time (min) Flow (ml / min) Solvent B 0.02 1.5 1.0 4, 90 85, 0 4, 91 1.0 5, 00 Stop 1.0

LC-MS method: LC-MS method G

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The UPLC analysis was performed on an Acquity UPLC CSH C18 column (50 mm x 2.1 mm) Packing I.D. 1.7 μm at 40 ° C.

The solvents used were:

A = 0.1% solution of formic acid in water v / v

B = 0.1% solution of formic acid in acetonitrile v / v

The gradient used was:

Time (min) Flow rate (ml / min) %A %IN 0 one 97 3 1.5 one 5 95 1.9 one 5 95 2.0 one 97 3

UV detection was a total signal at wavelengths from 210 nm to 350 nm.

Injection volume: 0.5 μL.

MC conditions

MS: Waters ZQ

Ionization mode: alternating positive and negative electrospray scan.

LC-MS method: LC-MS method H

UPLC analysis was performed on an Acquity UPLC CSH C18 column (50 mm x 2.1 mm) with a 1.7 μm internal diameter of the package at 40 ° C.

The solvents used were:

A = 10 mM ammonium bicarbonate in water, adjusted to pH 10 with ammonia solution.

B = acetonitrile.

The gradient used was:

Time (min) Flow rate (ml / min) %A %IN 0 one 97 3 0.05 one 97 3 1.5 one 5 95 19 one 5 95 twenty one 97 3

UV detection was a total signal at wavelengths from 210 nm to 350 nm.

Injection volume: 0.3 μL.

MC conditions

MS: Waters ZQ

Ionization mode: alternating positive and negative

LC-MS method: LC-MS method I

UPLC analysis was performed on an Acquity UPLC CSH C18 column (50 mm x 2.1 mm) with a 1.7 μm internal diameter of the package at 40 ° C.

The solvents used were:

A = 10 mM ammonium bicarbonate adjusted to pH 10 with 25% ammonium hydroxide solution.

B = acetonitrile

The gradient used was:

Time (min) Flow rate (ml / min) %A %IN 0 one 97 3 0.05 one 97 3 1.5 one 5 95 1.9 one 5 95 2.0 one 97 3

UV detection was a total signal at wavelengths from 210 nm to 350 nm.

Injection volume: 0.5 μL.

MC conditions

MS: Waters Acquity SQD or QDa Mass Detector

Ionization mode: alternating positive and negative

LC-MS method: LC-MS method J

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UPLC analysis was performed on an Acquity UPLC CSH C18 column (50 mm x 2.1 mm) with a 1.7 μm internal diameter of the package at 40 ° C.

The solvents used were:

A = 0.1% solution of formic acid in water v / v

B = 0.1% solution of formic acid in acetonitrile v / v

The gradient used was:

Time (min) Flow rate (ml / min) %A %IN 0 one 97 3 1.5 one 5 95 1.9 one 5 95 2.0 one 97 3

UV detection was a total signal at wavelengths from 210 nm to 350 nm.

Injection volume: 0.5 μL.

MC conditions

MS: Waters Acquity SQD or QDa Mass Detector

Ionization mode: alternating positive and negative

LC-MS method: LC-MS method K

UPLC analysis was performed on an Acquity UPLC CSH C18 column (50 mm x 2.1 mm) with a 1.7 μm internal diameter of the package at 40 ° C.

The solvents used were:

A = 0.1% solution of TPA in water v / v

B = 0.1% solution of TPA in acetonitrile v / v

The gradient used was:

Time (min) Flow rate (ml / min) %A %IN 0 one 97 3 1.5 one 5 95 1.9 one 5 95 2.0 one 97 3

UV detection was a total signal at wavelengths from 210 nm to 350 nm.

Injection volume: 0.5 μL.

MC conditions

MS: Waters Acquity SQD or QDa Mass Detector

Ionization mode: alternating positive and negative

LC-MS method: LC-MS method L

Device

LC: Waters Acquity Binary Solvent Manager Column Manager 55C Autosampler: CTC Leap PAL 3 UV: Waters Acquity PDA (210-360 nm) ELS Sedere Sedex 85C (45C) MS: mass detector Waters Acquity SQD Polarity (positive or negative); mode (continuum); scan time (10 Hz); capillary B (0.8); cone B (12); Column: Thermo Hypersil Gold (C18, 20x2.1 mm, particle diameter 1.9 μm) Solvent A: H ₂ O, 0.02% TPA Solvent B: MeCN, 0.02% TPA

Gradient Time (min) Flow (ml / min) Solvent B 0.02 16 0.5 1.90 95 1.91 0.5 2.00 Stop

LC-MS method: LC-MS method M

LC-MS analysis was performed on a Waters Sunfire C18 column (50 mm x 3.0 mm internal package diameter 5 μm) at room temperature on an Agilent 1200 HPLC with a Model 6140 Quad mass spectrometer.

The solvents used were: A = 0.1% solution of TPA in water, v / v. B = 0.1% solution of TPA in acetonitrile v / v

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The gradient used was:

Time Sweat to %A %IN 0 1 ml 90 ten 2.5 1 ml 0 one hundred 4.2 1 ml 0 one hundred

UV detection wavelength (bandwidth 8): 220 nm and 254 nm.

Injection volume: 1 μL

MC conditions

MS: Agilent 6140 Quad MS

Ionization mode: positive

LC-MS method: LC-MS method N

LC-MS analysis was performed on an Agilent Zorbax XDB-C18 (150 mm x 4.6 mm package inner diameter 5 μm) at room temperature on an Agilent 1200 HPLC with a Model 6140 Quad MS mass spectrometer.

The solvents used were: A = 0.1% solution of TPA in water, v / v. B = 0.1% solution of TPA in acetonitrile v / v

The gradient used was:

Time Sweat to %A %IN 0 1 ml 90 ten 12 1 ml 0 one hundred 13 1 ml 0 one hundred

UV detection wavelength (bandwidth 8): 220 nm and 254 nm.

Injection volume: 1 μL

MC conditions

MS: Agilent 6140 Quad MS Ion mode: positive

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The following abbreviations may be used in this description:

Abbreviated designation Value Ace OH acetic acid aq. water VVg ₃ boron tribromide BOC, tBOC tert-butoxycarbonyl brine saturated salt solution BuOH butanol CDC1 ₃ deuterated chloroform GDI 1,1'-carbonyldiimidazole CH ₂ C1 ₂ or DCM methylene chloride or dichloromethane CH ₃ CN or MeCN acetonitrile CH3NH2 methylamine d day DAST diethylaminosulfur trifluoride DOE 1,2-dichloroethane DIEA or DIPEA diisopropylethylamine DMA dimethylacetamide DMAP 4-dimethylaminopyridine DMF N, N-dimethylformamide DMSO dimethyl sulfoxide EDO 1-ethyl-3- (3- dimethylaminopropyl) carbodiimide equiv eq Et ethyl Et ₃ N or TEA triethylamine Et ₂ O diethyl ether EtOAc ethyl acetate EtOH ethanol ECO columnar flash romatography h, hr h (s) HATU hexafluorophosphate 0- (7-azabenzotriazole-1yl) -Ν, Ν, Ν ', N'-tetramethyluronium HCl hydrochloric acid HOAt 1-hydroxy-7-azabenzotriazole HOBt hydroxybenzotriazole HPLC high performance liquid chromatography IC1 iodine monochloride

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IPA isopropyl alcohol i-Pr ₂ NEt Ν ', N'-diisopropylethylamine K ₂ CO ₃ potassium carbonate KHMDS potassium bis (trimethylsilyl) amide KOt-Bu potassium tert-butoxide KOH potassium hydroxide lx-ms liquid chromatography-mass spectroscopy LiAlH ₄ lithium aluminum hydride L1HDMS lithium hexamethyldisilazide LiOH lithium hydroxide Me methyl MeOH or CH ₃ OH methanol MgSO ₄ magnesium sulfate min min (s) MS mass spectrum pw microwave NaBH ₄ sodium borohydride Na ₂ CO ₃ sodium carbonate NaHCO ₃ bicarbonate of soda NaOH sodium hydroxide Na ₂ SO ₄ sodium sulfate NBS Ν-bromosuccinimide N ₂ H ₂ hydrazine nh ₄ ci ammonium chloride nh ₄ oh ammonium hydroxide NiCl ₂ -6H ₂ O nickel (11) chloride hexahydrate NMR N-methyl-2-pyrrolidone NMR nuclear magnetic resonance Pd / C palladium on charcoal Ph phenyl POC1 ₃ phosphoryl chloride PSI pound-force per square inch RB round bottom rm or rxn mixture reaction mixture rt / RT room temperature satd. saturated sm starting material TBAF tetra-n-butylammonium fluoride TEA trifluoroacetic acid THE tetrahydrofuran TMEDA tetrame thylethylenediamine TMSI trimme tilsilyl iodide TMSN ₃ trimethylsilyl azide

TZR 2,4,6-tripropyl-1,3,5,2,4,6- trioxatriphosphorinan-2,4,6-trioxide t _R or Rf or R _t Hold Time TsOH p-toluenesulfonic acid

Intermediate connection 1

Stage 1:

4-chloro-3-methoxy-5-nitrobenzamide

Methyl 4-chloro-3-methoxy-5-nitrobenzoate (1000 mg, 4.07 mmol) was stirred in NH ₄ OH (10 ml, 7 7 mmol) at room temperature for 24 h. Then the reaction temperature was raised to 50 ° C for 2 hours. An additional 2 ml (about 3.7 equiv.) NH4OH was added to the vessel. After additional stir

- 34 036435 for 2 h at 50 ° C (total 4 h), the reaction mixture was cooled to room temperature. The solid was filtered off and washed with cold water. The solid was dried in vacuo and lyophilized to give 4-chloro-3-methoxy-5-nitrobenzamide (710 mg, 2.99 mmol, 73% yield) as a tan solid. 1H NMR (400 MHz, DMSOA) δ ppm 8.31 (br s, 1H), 8.06 (d, J = 1.77 Hz, 1H), 7.88 (d, J = 1.77 Hz, 1H, 7.81 (br s, 1H), 4.02 (s, 3H). LCMS (LCMS Method D): retention time = 0.71 min, [M + H] + = 230, nine.

Stage 2: 4-chloro-3-hydroxy-5-nitrobenzamide

4-Chloro-3-methoxy-5-nitrobenzamide (1 g, 4.34 mmol) was suspended in dry DCM (15 ml) and stirred at room temperature. BBr ₃ (17.4 ml, 1 M solution in DCM) was added dropwise to the reaction mixture. A slurry formed quickly and was stirred overnight at room temperature under nitrogen. The reaction mixture was poured into ice water (300 ml) and stirred vigorously for 30 minutes. The resulting slurry was filtered and the solids dried to give the title compound (610 mg, 2.82 mmol, 65% yield). ¹ H NMR (400 MHz, DMSO-06) δ; ppm 11.53 (br s, 1H), 8.17 (br s, 1H), 7.92 (s, 1H), 7.72 (s, 1H), 7.66 (br s, 1H ). LCMS (LCMS Method D): retention time = 0.60 min, [M + H] ⁺ = 217.

Intermediate connection 2

4- (5- (5-Carboxy-3-methyl-1 N-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1 N-pyrazole-5-carboxylic acid

Stage 1:

Ethyl 3-methyl-1- (5 - (trimethylsilyl) pent-4-yn-1 -yl) -1 H-pyrazole-5-carboxylate

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A mixture of ethyl 3-methyl-1Н-pyrazole-5-carboxylate (22 g, 143 mmol), (5-chloropent-1-yn-1-yl) trimethylsilane (24.94 g, 143 mmol), K ₂ CO ₃ ( 39.4 g, 285 mmol) and DMF (4 ml) were stirred at 60 ° C overnight under nitrogen gas. The mixture was then dissolved in DCM and washed with water. The organic phase was dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure and purified by column chromatography on silica gel (petroleum ether / EYOAc = 10: 1) to give ethyl 3-methyl-1- (5- (trimethylsilyl) pent- 4-yn-1-yl) -1H-pyrazole-5-carboxylate (12.5 g, 42.7 mmol, 30% yield) as a colorless oil. LCMS (LCMS Method A): retention time = 2.43 min, [M + H] + = 293.

Stage 2:

Ethyl 3-methyl-1 - (pent-4-yn-1-yl) -1H-pyrazole-5-carboxylate ° S

Me

A mixture of ethyl 3-methyl-1- (5- (trimethylsilyl) pent-4-yn-1-yl) -1H-pyrazole-5-carboxylate (37.7 g, 129 mmol), K ₂ CO ₃ (44.5 g, 322 mmol) and EtOH (800 ml) were stirred at room temperature overnight. The mixture was then filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM, washed with water, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give ethyl 3-methyl-1- (pent-4-yn-1-yl) -1H-pyrazole-5carboxylate (20 d, 91 mmol, yield 70.4%) as a colorless oil. LCMS (LCMS Method A): retention time = 2.08 min, [M + H] + = 221.

Stage 3:

Benzyl 1-ethyl-3-methyl-1H-pyrazole-5-carboxylate

About ^ -Me | „

Me

A mixture of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (20 g, 130 mmol), (bromomethyl) benzene (22.2 g, 130 mmol), K ₂ CO ₃ (26.9 g, 195 mmol) and DMF (200 ml) were stirred at 60 ° C overnight. The mixture was then dissolved in DCM, washed with water, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / EtOAc = 10: 1) to give benzyl 1-ethyl-3-methylpyrazole -5-carboxylate (31.4 g, 129 mmol, 99% yield) as a colorless oil. LCMS (LCMS Method A): retention time = 2.09 min, [M + H] + = 245.

Stage 4:

Benzyl 1-ethyl-4-iodo-3-methyl-1H-pyrazole-5-carboxylate

A mixture of benzyl 1-ethyl-3-methyl-1H-pyrazole-5-carboxylate (31.6 g, 129 mmol), 1-iodopyrrolidine-2.5 dione (34.9 g, 155 mmol) and DMF (400 ml) was stirred at 90 ° C for 2 days. The mixture was then allowed to cool to room temperature, dissolved in DCM and washed with saturated aqueous sodium thiosulfate solution. The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / EYOAc = 10: 1) to give benzyl 1-ethyl-4-iodo-3-methyl-1H- pyrazole-5carboxylate (42.6 g, 115 mmol, 89% yield). LCMS (LCMS Method A): holding time = 2.31 min, [M + H] + = 371.

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Stage 5:

Benzyl 4- (5 - (5- (ethoxycarbonyl) -3-methyl-1 H-pyrazol-1-yl) pent-1-yn-1-yl) -1-ethyl-3-methyl- 1H-pyrazole-5 -carboxylate

EY

Me

Pd (PPh ₃ ) ₂ CI ₂ , Cui

TEA, 60 ° C

Me

A mixture of ethyl 3-methyl-1- (pent-4-yn-1-yl) -1H-pyrazole-5-carboxylate (10.0 g, 45.4 mmol), benzyl 1ethyl-4-iodo-3-methyl- 1Н-pyrazole-5-carboxylate (16.8 g, 45.4 mmol), copper (I) iodide (0.864 g, 4.54 mmol), bis (triphenylphosphine) palladium (II) chloride (0.319 g, 0.454 mmol) and Et ₃ N (200 ml) was stirred at 60 ° C overnight under nitrogen gas. The mixture was then dissolved in DCM and washed with water. The organic phase was dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure and purified by column chromatography on silica gel (petroleum ether / EuAc = 5: 1) to give benzyl 4- (5- (5- (ethoxycarbonyl) -3- methyl 1H-pyrazol-1-yl) pent-1-yn-1yl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylate (9.5 g, 20.5 mmol, 45.3% yield ) as a yellow solid. LCMS (LCMS Method B): retention time = 2.66 min, [M + H] + = 463.

Stage 6:

4- (5 - (5- (Ethoxycarbonyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5carboxylic acid

A mixture of benzyl 4- (5 - (5- (ethoxycarbonyl) -3-methyl-1H-pyrazol-1-yl) pent-1-yn-1-yl) -1-ethyl-3-methyl1H-pyrazole-5-carboxylate (19.0 g, 41.10 mmol), 10% Pd / C (0.22 g, 2.05 mmol) and THF (500 ml) were stirred at room temperature in an atmosphere of hydrogen gas (4 atm) for 2 days ... Then, the reaction mixture was filtered and concentrated under reduced pressure. The resulting residue was recrystallized from EtOAc / petroleum ether (1: 5, v / v) to give 4- (5- (5 (ethoxycarbonyl) -3-methylpyrazol-1-yl) pentyl) -1-ethyl-3- methylpyrazole-5-carboxylic acid (10.5 g, 27.90 mmol, 67.9% yield). ¹ H NMR (400 MHz, CDC13) δ ppm 6.63 (s, 1H), 4.57-4.48 (m, 4H), 4.38-4.32 (m, 2H), 2.74- 2.62 (m, 2H), 2.32 (s, 3H), 2.23 (s, 3H), 1.91-1.86 (m, 2H), 1.59-1.54 (m, 2h), 1.45-1.37 (m, 8h). LCMS (LC-MS Method A): retention time = 1.59 min, [M + H] + = 377.

Stage 7:

4-4- (7- (5-Carboxy-3-methyl-1 N-pyrazol-1-yl) heptyl) -1-ethyl-3-methyl-1 N-pyrazole-5-carboxylic acid

To a suspension of 4- (5 - (5- (ethoxycarbonyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (9.0 g , 23.9 mmol) in MeOH (120 ml) and water (120 ml), stirring at room temperature, was added 2 M aqueous NaOH solution (60 ml, 119.5 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The mixture was then acidified to pH 4 with the addition of 6 M HCl solution, and a solid precipitated from the reaction mixture. The solid was collected by filtration and dried under reduced pressure to give 4- (5- (5carboxy-3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (6.5 g, 18.7 mmol, 78.1% yield) as a white solid. ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 6.57 (s, 1H), 4.40-4.34 (m, 4H), 2.53 (t, J = 7.6 Hz, 2H) , 2.16 (s, 3H), 2.09 (s, 3H), 1.74-1.67 (m, 2H), 1.44-1.37 (m, 2H), 1.27-1 , 16 (m, 5H). LCMS (LCMS Method A): retention time = 1.40 min, [M + H] + = 349.

Intermediate 3 (3-Bromopropoxy) (tert-butyl) dimethylsilane

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Br

HE

Br

Si O

TBDMSCI | ^x imidazole

To 1H-imidazole (13.4 g, 197 mmol) in DCM (100 ml) was added 3-bromopropan-1-ol (13.7 g, 99 mmol), then slowly tert-butylchlorodimethylsilane (17.8 g, 118 mmol ) in DCM (20 ml). After 3 h at room temperature, the reaction mixture was concentrated to a volume of about 100 ml and poured into EtOAc (800 ml), washed with 5% aqueous citric acid solution (2 x 200 ml) and brine. The organic layer was dried over MgSO ₄ , filtered and concentrated to give the title compound (10.0 g, 39.5 mmol, 40% yield). 1H NMR (400 MHz, chloroform-d) δ ppm 3.78 (t, J = 5.70 Hz, 2H), 3.56 (t, J = 6.46 Hz, 2H), 2.07 (t, J = 5.83 Hz, 2H), 0.94 (s, 9H), 0.11 (s, 6H).

Intermediate connection 4

2,2,3,3-Tetrafluorobutane-1,4-diamine

Stage 1:

2,2,3,3-tetrafluorobutane-1,4-diyl bis (4-methylbenzenesulfonate)

To 2,2,3,3-tetrafluorobutane-1,4-diol (10.0 g, 61.7 mmol) in pyridine (150 ml) at 0 ° C was added 4methylbenzene-1-sulfonyl chloride (29.4 g, 154 mmol) for 5 min, and then the reaction mixture was heated to 55 ° C. After 1 day, the reaction was quenched with ice water and the resulting solid was collected by filtration, dissolved in DCM (200 ml) and washed with 5% aqueous H ₂ SO ₄ (100 ml x 3). The organic layer was dried over Na ₂ SO ₄ and concentrated to give the title compound (27.3 g, 58.0 mmol, 94% yield) as a white solid. LC-MS (LC-MS Method A): retention time = 1.750 min, [M + H] + = 470.9

Stage 2:

1,4-diazido-2,2,3,3-tetrafluorobutane

2,2,3,3-Tetrafluorobutane-1,4-diyl bis (4-methylbenzenesulfonate) (10.0 g, 21.3 mmol) and sodium azide (5.53 g, 85.0 mmol) in DMF ( 40 ml) was stirred at 110 ° C overnight. The reaction was quenched with NaClO (aq) and extracted with DCM (5 ml x 3). The combined organic layers were washed with water (10 ml), dried over Na2SO4 and concentrated to give the title compound (3.5 g, 16.5 mmol, 78% yield). LCMS (LCMS Method A): retention time = 1.520 min, [M + H] ⁺ = 213.1.

Stage 3:

2,2,3,3-tetrafluorobutane-1,4-diamine

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To a solution of 1,4-diazido-2,2,3,3-tetrafluorobutane (36.0 g, 170 mmol) in MeOH (350 ml) was added

10% Pd on carbon (18.1 g, 17.0 mmol). The reaction mixture was stirred at 40 ° C under a hydrogen atmosphere (4 atm) for 16 h. The mixture was filtered through a pad of celite, washed with MeOH and the filtrate was concentrated in vacuo to give the title compound (22.0 g, 124 mmol, 73% yield ). ¹ H

NMR (400 MHz, chloroform-d) δ ppm 3.12-3.37 (m, 4H), 1.43 (brs, 4H).

Intermediate 5:

-Ethyl-3-methyl-1 H-pyrazole-5-carbonyl isothiocyanate

To a 1 L round bottom flask was added 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (25 g, 162 mmol) and DCM (500 ml). To this heterogeneous solution was added DMF (0.1 ml, 1.291 mmol) followed by the slow addition of oxalyl chloride (15.61 ml, 178 mmol). Bubbling was noted during addition. After stirring for 1 hour at room temperature, the volatiles were removed in vacuo and the crude product was evaporated twice with dichloromethane (100 ml each time). It was assumed that there was 100% yield, and the crude 1-ethyl-3-methyl-1H-pyrazole 5-carbonyl chloride (28.0 g, 162 mmol, 100% yield)) was used directly as such in the next reaction.

KSCN (18.92 g, 195 mmol) and acetone (463 mL) were added to a dry 1 L round bottom flask. This clear homogeneous solution was cooled to 0 ° C. 5 min after stirring at 0 ° C, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride (28 g, 162 mmol) was added as a solution in acetone (25 ml). After the addition was complete, the reaction mixture was stirred at 0 ° C. After 1 min, additional KSCN (ca. 2 g) was added and the reaction was stirred for an additional 20 min. At this time, hexane (200 ml) was added to the reaction mixture and the crude heterogeneous solution was concentrated in vacuo to one third of the volume. The process of adding and concentrating hexane was repeated twice (300 ml of hexane each time). After the last concentration, hexane (200 ml) was added and the solid was removed by filtration, washed with hexane (100 ml). The resulting clear light yellow filtrate was concentrated and purified by chromatography (330 g Gold columns with silica gel, eluting with 0-20% EtOAc / hexane). The desired product elutes at about 7% EtOAc / hexane. The desired fractions were combined and concentrated to give 1-ethyl-3-methyl-1H pyrazole-5-carbonyl isothiocyanate (27.5 g, 139 mmol, 86% yield) as a clear colorless liquid. 1H NMR (400 MHz, chloroform-d) δ ppm 6.77 (s, 1H), 4.54 (q, J = 7.10 Hz, 2H), 2.34 (s, 3H), 1.44 ( t, J = 7.22 Hz, 3 H); LCMS (LCMS Method D): retention time = 1.16 min, [M + H] + = 196.1. The product acyl isothiocyanate degrades over time and therefore an approximately 0.4 M solution in 1,4-dioxane was prepared and frozen in order to avoid / was slow decomposition. This solution was thawed and used directly in subsequent reactions.

Intermediate 6:

(E) -1- (4-Aminobut-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide hydrochloride

N-BOC H

VOS

- 39 036435

Stage 1:

(E) -tert-butyl (4 - ((4-carbamoyl-2-nitrophenyl) amino) but-2-en-1-yl) carbamate

A mixture of 4-fluoro-3-nitrobenzamide (10.0 g, 54.3 mmol), (E) -tert-butyl (4-aminobut-2-en-1yl) carbamate (10.62 g, 57.0 mmol) and K2CO3 (15.01 g, 109 mmol) in DMSO (200 ml) was stirred at room temperature overnight. The reaction mixture was poured into water (2000 ml) and stirred for 30 minutes. The resulting solid was collected by filtration to give the title compound (18.3 g, 52.2 mmol, 96% yield). LCMS (LCMS Method A): retention time = 1.38 min, [2M + H] + = 700.5.

Stage 2:

(E) -tert-butyl (4 - ((2-amino-4-carbamoylphenyl) amino) but-2-en-1-yl) carbamate

K (E) -tert-butyl (4 - ((4-carbamoyl-2-nitrophenyl) amino) but-2-en-1-yl) carbamate (18.3 g, 52.2 mmol) in DMF (300 ml ) was added stannous chloride dihydrate (58.9 g, 261 mmol). After stirring at room temperature overnight, the reaction mixture was added dropwise to saturated NaHCO ₃ solution (2000 ml) and was extracted with EtOAc (5 x 500 ml). The combined organic layers were washed with brine (200 ml), dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (16.5 g, 51.5 mmol, 99% yield) as a yellow oil. LCMS (LCMS Method A): retention time = 1.275 min, [M-BOC + H] + = 221.1.

Stage 3:

(E) -tert-butyl (4- (2-amino-5-carbamoyl-1H-benzo | imidazol-1-yl) but-2-en-1-yl) carbamate

A mixture of (E) -tert-butyl (4- ((2-amino-4-carbamoylphenyl) amino) no-2-en-1-yl) carbamate (16.5 g, 51.5 mmol) and cyanogen bromide (8, 18 g, 77 mmol) in THF (200 ml) was heated under reflux overnight. The reaction mixture was cooled to room temperature, diluted with saturated aqueous NaHCO ₃ (500 ml) and extracted with EtOAc (5 x 300 ml). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified on silica gel eluting with DCM in MeOH 50: 1 to 20: 1 (+ 3% NH4OH) to afford the title compound (13.7 g, 39.7 mmol, 77% yield) as an off-white solid substances. LCMS (LCMS Method A): retention time = 1.150 min, [M + H] + = 346.1.

Stage 4:

(E) -tert-butyl (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [d] imidazol-1-yl) but-2-ene -1-yl) carbamate

- 40 036435

To 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (9.17 g, 59.5 mmol) in DCM (500 ml) at 0 ° C was added EDC (20.53 g, 107 mmol) and HOBT (18.22 g, 119 mmol). After 15 min, a mixture of (E) tert-butyl (4- (2-amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) carbamate (13.7 g , 39.7 mmol) in DMF (50 ml), followed by the addition of TEA (27.6 ml, 198 mmol). The reaction mixture was warmed to room temperature, stirred overnight and concentrated. The residue was diluted with water (500 ml) and extracted with ethyl acetate (3 x 300 ml) and the combined organic phases were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified on silica gel eluting with DC: MeOH 50: 1 to 20: 1 to give the crude product, which was washed with DCM (300 ml) and collected by filtration to give the title compound (14.0 g, 29.1 mmol, yield 73%) as an off-white solid. ¹ H NMR (400 MHz, DMSO-ab) δ ppm 12.84 (s, 1H), 8.00-7.97 (m, 2H), 7.80-7.78 (m, 1H), 7, 49 (d, J = 8.4 Hz, 1 H), 7.34 (s, 1H), 6.95 (t, J = 5.5 Hz, 1H), 6.66 (s, 1H), 5 , 73-5.65 (m, 2H), 4.83 (d, J = 4.3 Hz, 2H), 4.62 (q, J = 7.0 Hz, 2H), 3.52 (s, 2H), 2.18 (s, 3H), 1.38-1.33 (m, 12H); LCMS (LCMS Method A): retention time = 1.4 09 min, [M + H] + = 482.0.

Stage 5:

(E) -1 - (4-aminobut-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzoI imidazole 5-carboxamide hydrochloride

To a suspension of (E) -tert-butyl (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo ^] imidazol-1-yl) but-2- en-1-yl) carbamate (3.00 g, 6.23 mmol) in dioxane (60 ml) was added 4N HCl in dioxane (15.6 ml, 62.3 mmol) followed by the addition of MeOH (15 ml) to dissolve some remaining solid. After 30 minutes at room temperature, the reaction mixture became cloudy and was stirred for about 3 days. The resulting solid was collected by filtration and washed with DCM to give the title compound (2.0 g, 4.8 mmol, 77% yield) as a white solid. ¹ H NMR (400 MHz, DMSO ^ 6) δ ppm 7.97-8.09 (br s, 1H), 7.82 (d, J = 8.11 Hz, 1H), 7.50 (d, J = 8.11 Hz, 1H), 7.38 (brs, 1H), 6.70 (s, 1H), 5.97-6.08 (m, 1H), 5.68-5.80 (m, 1H), 4.91 (d, J = 4.31 Hz, 2H), 4.60 (q, J = 6, 67 Hz, 2H), 3.42 (brs, 2H), 2.18 (s, 3H), 1.36 (t, J = 6.97 Hz, 3H); LCMS (LCMS Method D): retention time = 0.53 min, [M + H] + = 382.2.

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Intermediate connection 7

1- (5- (5- (Ethoxycarbonyl) -1-ethyl-3-methyl-1H-pyrazol-4-yl) pentyl) -3-methyl-1H-pyrazole-5carboxylic acid

Stage 1:

Benzyl 3-methyl-1 H-pyrazole-5-carboxylate

A mixture of 3-methyl-1H-pyrazole-5-carboxylic acid (50 mg, 0.396 mmol) and KHCO3 (47.6 mg, 0.476 mmol) in DMSO (2 ml) was stirred for 30 min and (bromomethyl) benzene (0.045 ml, 0.377 mmol). The mixture was stirred for 4 h at room temperature, diluted with EtOAc (20 ml), washed with water and brine, and dried over Na ₂ SO ₄ . The mixture was filtered and concentrated, and the residue was purified by column chromatography (Combiflash, 0-50% EtOAc in hexane) to give the title compound (66 mg, 0.305 mmol, 77% yield) as a white solid. ¹ H NMR (400 MHz, DMSOC) δ ppm 13.19 (br s, 1H) 7.34-7.48 (m, 5H), 6.52 (s, 1H), 5.29 (s, H 2H) 2.27 (s, 3H). LCMS (LCMS Method D): retention time = 0.86 min, [M + H] + = 216.9.

Stage 2:

Benzyl 3-methyl-1 - (pent-4-yn-1-yl) -1 H-pyrazole-5-carboxylate

Me

A mixture of DIAD (25.9 ml, 133 mmol) and triphenylphosphine (34.9 g, 133 mmol) in tetrahydrofuran (THF) (600 ml) was stirred for 30 min at 0 ° C and pent-4-yn-1- ol (11.36 ml, 122 mmol). The mixture was stirred for 30 min and benzyl 3-methyl-1H-pyrazole-5-carboxylate (24 g, 111 mmol) was added. The mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was diluted with EtOAc (1000 ml), washed with saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The oily residue was treated with 10% EtOAc in hexane (500 ml) and a white precipitate formed. The precipitate was filtered off and washed with 10% EtOAc in hexane. The combined filtrates were concentrated and the residue was purified by column chromatography (Combiflash, 0-15% EtOAc in hexanes) to afford the title compound (27.5 g, 97 mmol, 88% yield) as a white solid. ¹ H NMR (400 MHz, chloroform-d) δ ppm 7.34-7.47 (m, 5H), 6.68 (s, 1H) 5.33 (s, 2H), 4.63 (t, J = 7.03 Hz, 2H), 2.30, 3H), 2.19-2.26 (m, 2H), 2.09 (quint, J = 7.09 Hz, 2H), 1.97 (br s, 1H); LCMS (LCMS Method D): retention time = 1.21 min, [M + H] + = 283.0.

Stage 3:

Ethyl 1-ethyl-3-methyl-1 H-pyrazole-5-carboxylate

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Oxalyl chloride (5.68 ml, 64.9 mmol) was added to a suspension of 1-ethyl-3-methyl-1H-pyrazole-5 carboxylic acid (5 g, 32.4 mmol) in DCM (40 ml) at room temperature under N2 and two drops of DMF were added. The mixture was stirred for 2 hours at room temperature, concentrated and dried in vacuo. Ethanol (50 ml, 856 mmol) was added and the mixture was stirred for 1 hour at room temperature. The reaction mixture was concentrated and dried in vacuo to give a light yellow oil which was taken up in EtOAc (100 ml), washed with saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ , filtered, concentrated and the resulting residue was dried in vacuo with obtaining the title compound (5.5 g, 30.2 mmol, 93% yield) as a light yellow oil. Ή NMR (400 MHz, chloroform-d) δ ppm 6.63 (s, 1H) 4.56 (q, J = 7.11 Hz, 2H), 4.35 (q, J = 7.11 Hz, 2H ), 2.30 (s, 3H) 1.44 (t, J = 7.2 Hz, 3H), 1.39 (t, J = 7.2 Hz, 3H). LCMS (LC-MS Method E): retention time = 0.81 min, [M + H] + = 183.1.

Stage 4:

Ethyl 1-ethyl-4-iodo-3-methyl-1H-pyrazole-5-carboxylate

A mixture of ethyl 1-ethyl-3-methyl-1H-pyrazole-5-carboxylate (5.5 g, 30.2 mmol) and NIS (8.15 g, 36.2 mmol) in DMF (100 ml) was heated to 90 ° C and stirred for 3 days in an atmosphere of N2. The reaction mixture was cooled to room temperature, diluted with EtOAc (200 ml), washed with saturated Na ₂ S ₂ O ₃ solution, 5% LiCl solution and brine, dried over Na ₂ SO ₄ , filtered, concentrated and the resulting residue was purified by column chromatography ( Combiflash, 0-7% EtOAc in hexane) to afford the title compound (9.1 g, 29.5 mmol, 98% yield) as a colorless oil. 1H NMR (400 MHz, chloroform-d) δ ppm 4.57 (q, J = 7.03 Hz, 2H), 4.43 (q, J = 7.03 Hz, 2H), 2.32 (s, 3H) 1.45-1.50 (m, 3H) 1.39-1.45 (m, 3H). LCMS (LCMS Method D): retention time = 1.12 min, [M + H] ⁺ = 308.9.

Stage 5:

-Ethyl 4- (5- (5 - ((benzyloxy) carbonyl) -3 -methyl-1 H-pyrazol-1 -yl) pent-1-yn-1 -yl) -1-ethyl-3-methyl1H-pyrazole -5-carboxylate

The flask, which was pre-purged with nitrogen, was charged with Cs ₂ CO ₃ (23.08 g, 70.8 mmol), 1.10 phenanthroline (1.915 g, 10.63 mmol), copper (1) chloride (0.175 g, 1.771 mmol) , benzyl 3-methyl-1- (pent-4-yn1-yl) -1H-pyrazole-5-carboxylate (10 g, 35.4 mmol), ethyl 1-ethyl-4-iodo-3-methyl-1H- pyrazole-5-carboxylate (13.10 g, 42.5 mmol), Pd [P (o-tolyl) ₃ ] ₂ (0.760 g, 1.063 mmol) and degassed toluene (100 ml). The mixture was degassed for 15 min, heated to 100 ° C and stirred overnight (18 h) under N2. The reaction mixture was cooled to room temperature and diluted with EtOAc. Inorganic solids were filtered off and washed with EtOAc. The combined organic phases were concentrated and the residue was purified by silica gel chromatography (EtOAc / hexane 0-25%) to afford the title compound (11.38 g, 24.60 mmol, 69.5% yield) as a colorless oil. 1H NMR (400 MHz, chloroform-d) δ ppm 7.34-7.47 (m, 5H), 6.68 (s, 1H), 5.31 (s, 2H), 4.67 (t, J = 7.03 Hz, 2H), 4.51 (q, J = 7.19, Hz, 2H), 4.39 (q, J = 7.03 Hz, 2H), 2.51 (t, J = 7.28 Hz, 2H), 2.31 (s, 3H), 2.29 (s, 3H), 2.17 (s, t, J = 7.15 Hz, 2H), 1.40 (t, J = 7.03 Hz, 6H). LCMS (LCMS Method D): retention time = 1.43 min, [M + H] ⁺ = 463.3.

Stage 6:

1- (5- (5 - (Ethoxycarbonyl) -1-ethyl-3-methyl-1 N-pyrazol-4-yl) pentyl) -3 -methyl-1 N-pyrazole-5 carboxylic acid

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In a flask loaded with ethyl 4- (5- (5 - ((benzyloxy) carbonyl) -3-methyl-1H-pyrazol-1-yl) pent-1-yn-1yl) -1-ethyl-3-methyl-1H α-pyrazole-5-carboxylate (11.3 g, 24.43 mmol) and Pd / C (2.60 g, 2.443 mmol) ethanol (200 mL) was added. The flask was purged with N2, then with hydrogen (balloon) and the mixture was stirred under H ₂ overnight (18 hours). The catalyst was filtered off and the filtrate was concentrated in vacuo to afford the title compound (8.89 g, 23.62 mmol, 97% yield) as a white solid. ¹ H NMR (400 MHz, DMSO ^) δ ppm 13.14 (br s, 1H), 6.57 (s, 1H), 4.33-4.43 (m, 4H), 4.28 (m , J = 7.09 Hz, 2H) 2.51-2.56 (m, 2H), 2.16 (s, 3H), 2.10 (s, 3H) 1.72 (m, J = 7, 34 Hz, 2H) 1.41 (m, J = 7.58 Hz, 2H), 1.25-1.31 (m, 6H), 1.16-1.24 (m, 2H). LCMS (LCMS Method D): retention time = 1.07 min, [M + H] + = 377.2.

Example 1. l, l '- ((2R, 3R) -2,3-Dihydroxybutane-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H -benzo ^] imidazole-5-carboxamide) ditrifluoroacetate

Stage 1:

(4R, 5R) -4,5 -bis (azidomethyl) -2,2-dimethyl-1,3-dioxolane

A mixture of ((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene) bis (4-methylbenzenesulfonate) (3.874 g, 8.23 mmol) and sodium azide (1.338 g, 20.58 mmol) in DMF (20 ml) was heated at 8 ° C for 15 h. The reaction mixture was concentrated in vacuo to remove DMF and the residue was dissolved in DCM / water. The two-phase solution was transferred to a separatory funnel and the layers were separated. DCM layer

- 44 036435 Washed twice with water and once with brine, dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a pale yellow liquid (1.564 g, 7.37 mmol, 90% yield). 1H NMR (400 MHz, chloroform-d) δ ppm 4.10 (dd, J = 2.78, 1.26

Hz, 2H), 3.55-3.66 (m, 2H) 3.32-3.44 (m, 2H) 1.51 (s, 6H). LCMS (LCMS Method C): retention time = 0.8 9 min, [M + H] + = 214.0.

Stage 2:

((4R, 5 R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) dimethanamine

To a solution of (4R, 5R) -4,5-bis (azidomethyl) -2,2-dimethyl-1,3-dioxolane (1.561 g, 7.36 mmol) in dry THF (30 ml) at room temperature in an open round bottom flask with a capacity of 250 ml was added dropwise a 2 M solution of LiAlH ₄ in THF (3.68 ml, 7.36 mmol) over 10 min. The reaction mixture was then diluted with THF (30 ml) and the mixture was stirred for 30 min. The reaction was suppressed by adding dropwise to the reaction mixture 1.24 ml of a saturated aqueous solution of Na ₂ SO ₄ . After quenching, the yellow reaction mixture was stirred for 10 min and then allowed to settle. The resulting solid was removed by filtration, and the filtrate was dried over Na ₂ SO ₄ , filtered and concentrated to afford the title compound as a pale yellow oil (977 mg, 6.1 mmol, 83% yield). ¹ H NMR (400 MHz, chloroform-d) δ ppm 3.72-3.91 (m, 2H) 2.71-3.11 (m, 4H) 1.18-1.65 (m, 6H). LCMS (LC-MS Method C): retention time = 0.11 min, [M + H] + = 161.0.

Stage 3:

4,4 '- ((((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (azandiyl)) bis (3-nitrobenzamide)

A mixture of 4-fluoro-3-nitrobenzamide (2.233 g, 12.13 mmol), ((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5diyl) dimethanamine (0.9713 g, 6, 06 mmol) and K2CO3 (1.843 g, 13.34 mmol) in DMSO (20 ml) were stirred at 70 ° C for 90 min. The reaction mixture was cooled slightly and diluted with 200 ml of water. The resulting orange suspension was vigorously stirred for 60 min, isolated by filtration, and the filtered solid was dried on a Buchner funnel for 20 min. The slightly wet solid was transferred to a beaker containing Et ₂ O and the solid was further crushed with a spatula to remove excess water from the solid. The resulting solid was isolated by filtration, transferred to a 250 ml round bottom flask and dried for 3 days at 5b ° C in a vacuum oven to afford the title product (2.31 g, 4.73 mmol, 78% yield) as a yellow powder. 1H NMR (400 MHz, DMSO ^ 6) δ ppm 8.66 (d, J = 2.27 Hz, 2H), 8.50 (t, J = 5.56 Hz, 2H), 7.98-8, 10 (m, 4H), 7.34 (br s, 2H), 7.19 (d, J = 9.09 Hz, 2H), 4.22 (br s, 2H), 3.64-3 , 86 (m, 4H), 1.38 (s, 6H). LCMS (LCMS Method C): retention time = 0.7 8 min, [M + H] + = 489.2.

Stage 4:

4,4 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (azandiyl)) bis (3-aminobenzamide)

- 45 036435

A mixture of 4, 4 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (azandiyl)) bis (3nitrobenzamide) (2.293 g, 4.69 mmol) and 10% wet Pd / C (230 mg) in NMP (25 ml) in a 250 ml round-bottom flask with a reflux condenser, from which air was evacuated, and placed under balloon hydrogen for 18 h at room temperature, followed by heating to 80 ° C for 20 h. Then the reaction mixture was cooled and filtered through Celite®, rinsing with 4 ml of NMP. The filtrate containing the product was used directly in the next reaction as a solution in NMP. LCMS (LC-MS Method C): retention time = 0.50 min, [M + H] + = 429.2.

Stage 5:

1,1 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (2-amino-1H-benzo ^] imidazole 5- carboxamide) dihydrobromide

A solution of 4,4 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (azandiyl)) bis (3aminobenzamide) (1.0 g, 2.334 mmol) in NMP (16 ml) was treated with cyanogen bromide (0.618 g, 5.83 mmol) and the homogeneous reaction mixture was stirred at room temperature for 3 h. Additional cyanogen bromide (0.618 g) was added and the reaction mixture was stirred for 18 h at room temperature. Then added additional cyanogen bromide (1.236 g, 5.0 equiv) and the reaction mixture was stirred at room temperature. After 5.5 h the reaction mixture was heated at 72 ° C for 55 min, cooled and diluted with 160 ml EtOAc. The resulting suspension was stirred for 20 min and the solid was collected by filtration, rinsing with EtOAc. The resulting dark green hygroscopic solid was transferred to a vial and dried in a vacuum oven at 40 ° C for 3 days to afford the title product (1.35 g, 2.11 mmol, 90% yield) as a dark brown solid. substances. ¹ H NMR (400 MHz, DMSO 6) δ ppm: 12.99 (brs, 1H), 8.87 (brs, 3H), 8.10 (brs, 2H), 7.85-7 , 91 (m, 4H) 7.68 (d, J = 9.09 Hz, 2H), 7.49 (br s, 2H), 4.69-4.76 (m, 2H), 4.55 -4.63 (m, 2H), 4.36 (br s) 1.25 (s, 6H). LCMS (LCMS Method C): retention time = 0.40 min, [M + H] + = 479.2.

Stage 6:

1,1 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (2- (1-ethyl-3-methyl-1 N-pyrazole 5-carboxamido) -1H-benzo (^] imidazole-5-carboxamide) ditrifluoroacetate o

about

A mixture of 1,1 '- ((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (2-amino-1H-benzo ^] imidazole-5 -carboxamide) dihydrobromide (0.6647 g, 1.038 mmol), 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (0.32 g, 2.076 mmol), HATU (0.868 g, 2.284 mmol) and DIPEA (1.088 ml, 6.23 mmol) in NMP (4 ml) was heated at 140 ° C for 30 min in a microwave reactor. The reaction mixture was purified directly by reverse phase HPLC (Gilson®, 13-43% MeCN / 0.1% TFA in water, gradient 15 min, Luna column). The desired fractions were combined, concentrated in vacuo and placed under high vacuum for 15 h to afford the title product (14 0.0 mg, 0.143 mmol, 13.7% yield) as a dark green solid. 1H NMR (400 MHz, DMSO ^ 6) δ ppm 12.94 (br s, 2H), 7.98 (s, 4H), 7.66-7.76 (m, 2H), 7.47 (d , J = 8.59 Hz, 2H), 7.38 (br s, 2H), 6.86 (s, 2H), 4.75 (d, J = 10.11 Hz, 2H), 4.55 -4.96 (m, 6H), 4.40 (br s, 6H), 1.37 (t, J = 7.20 Hz, 6H), 1.14 (s, 6H). LCMS (LCMS Method C): retention time = 0.84 min, [M + H] ⁺ = 751.6.

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Stage 7:

1,1 '- ((2R, 3R) -2,3-dihydroxybutane-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1H-benzo ^] imidazole-5-carboxamide) ditrifluoroacetate

1,1 '- (((4R, 5R) -2,2-dimethyl-1,3-dioxolane-4,5-diyl) bis (methylene)) bis (2- (1-ethyl-3-methyl-1 N-pyrazole 5-carboxamido) -1H-benzo (^] imidazole-5-carboxamide) ditrifluoroacetate (113.8 mg, 0.116 mmol) was dissolved in formic acid (3.0 ml) and water (0.3 ml) and stirred at room temperature. temperature for 4 days. The reaction mixture was concentrated in vacuo at room temperature to give a green solid. The crude solid was dissolved in 1.4 ml DMSO and purified by HPLC (Gilson® Autoprep, Luna acid column, 5-35% MeCN: 0.1% in water TFA, 7 min gradient). The target fractions were concentrated in a vacuum and placed in a high vacuum for 3 days to obtain 1,1 '- ((2R, 3R) -2,3-dihydroxybutane-1,4-diyl) bis (2- (1-ethyl-3- methyl 1H-pyrazole-5-carboxamido) -1H-benzo (^] imidazole-5-carboxamide) ditrifluoroacetate (27 mg, 0.029 mmol, 24.7% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSO-d ^ δ ppm 12.89 (br s, 2H) 8.00 (s, 4H), 7.79 (d, J = 8.34 Hz, 2H), 7, 55 (d, J = 8.34 Hz, 2H), 4.51-4.67 (m, 4H), 4.27-4.47 (m, 4H), 4.09 (brs, 2H) , 5.50 (bs, 2H), 4.51-4.67 (m, 4H), 4.27-4.47 (m, 4H), 4.09 (bs, 2H), 2 09 (s, 6H), 1.32 (t, J = 7.07 Hz, 6H) LCMS (LCMS Method C): retention time = 0.67 min, [M + H] + = 711, 6.

Example 2. (E) -1,1 '- (But-2-ene-1,4-diyl) bis (2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H -benzo ^] imidazole-5-carboxamide)

TsOH, DCM

Grubbs-Honeyda II catalyst

1. to ₂ from ₃ , DMF

2. Zn / AcOH

3. BrCN, MeOH

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Stage 1:

-allyl-2-amino-1H-benzo ^] imidazole-5-carboxamide hydrobromide o

To a solution of 4-fluoro-3-nitrobenzamide (10.0 g, 54.3 mmol) in DMF (60 ml) was added allylamine (36, 6 ml, 489 mmol) dropwise at room temperature and the mixture was stirred for 5 min. At the end of this period, K ₂ CO ₃ (15.01 g, 109 mmol) was added in one portion and the mixture was stirred at room temperature for 30 minutes. Then DMF was removed in vacuo, the residue was suspended in 500 ml of water, the resulting orange precipitate was filtered off, washed with water and dried in vacuum.

The above precipitate was dissolved in AcOH (600.0 ml), the flask was placed in a water bath at 20 ° C and zinc (10.65 g, 163 mmol) was carefully added in small portions. The reaction was monitored by LC-MS and additional zinc (about 3 equiv) was added in small portions as needed until the reduction was complete. At the end of the reaction, as indicated by LC-MS, the solids were filtered off and the filtrate was concentrated in vacuo. The evaporation residue was taken up in DCM (500 ml) and EtOH (150 ml) and washed with 15% aqueous K ₂ CO ₃ solution (100 ml). The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo.

The above evaporation residue was dissolved in MeOH (200.0 ml), a 5.0 M solution of cyanogen bromide in CH3CN (11.95 ml, 59.7 mmol) was added in one portion and the mixture was stirred at room temperature for 18 hours. period, the reaction mixture was concentrated in vacuo, then redissolved in MeOH (200.0 ml). A mixture of toluene (100 ml) and CH ₃ CN (100 ml) was added and the resulting mixture was concentrated to dryness at 40 ° C (0-1 mbar) and dried in vacuo for 16 h to give 1-allyl-2-amino-1H- benzo ^] imidazole-5-carboxamide hydrobromide (11.3 g, 38.0 mmol, 70.0% yield) as a dark purple powder. Ή NMR (400 MHz, DMSOD ₆ ) δ ppm 8.83 (s, 2H), 8.07 (br s, 1H), 7.88 (d, J = 1.00 Hz, 1H), 7.82 (dd, 8.41, 1.38 Hz, 1H), 7.52 (d, J = 8.53 Hz, 1H), 7.43 (br s, 1H), 5.87-6.02 ( m, 1H), 5.25 (dd, J = 10.42, 0.88 Hz, 1H), 5.17 (dd, J = 17.32, 1.00 Hz, 1H), 4.84 (d , J = 5.02 Hz, 2H); LCMS (LCMS Method C): retention time = 0.38 min, [M + H] + = 216.9.

Stage 2:

1-Allyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide

1-allyl-2-amino-1H-benzo ^] imidazole-5carboxamide hydrobromide (2.5 g, 8.41 mmol), HATU (3.52 g, 9.25 mmol), 1 was loaded into a round-bottom flask with a capacity of 100 ml. -ethyl-3-methyl-1H-pyrazole-5carboxylic acid (1.427 g, 9.25 mmol) and NMP (25 ml). After stirring for 1 min at room temperature, DIPEA (7.33 ml, 42.1 mmol) was added and the mixture was stirred at room temperature for 40 h. At the end of this period, 2.0 ml of water was added and the mixture was stirred for 30 min. at room temperature. The mixture was then poured into 500 ml ice water and stirred vigorously for 1 hour. The dark purple solid was filtered off, brine (100 ml) was added and the next portion of the slightly lighter precipitate was filtered. The resulting transparent pink filtrate was kept at room temperature for 4 days, during which time the lightest pink precipitate formed from the solution. This final precipitate was filtered off, washed with water and air dried to give 1-allyl-2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide (1.88 g , 5.33 mmol, 63.4% yield) as a pale pink powder. T NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.84 (br s, 1H), 8.01 (s, 1H), 7.96 (br s, 1H), 7.78 (dd, J = 8.44, 1.59 Hz, 1H), 7.46 (d, J = 8.31 Hz, 1H), 7.32 (br s, 1H), 6.66 (s, 1H), 5 , 94-6.05 (m, 1H), 5.21 (dd, J = 10.27, 1.22 Hz, 1H), 5.15 (dd, J = 17.12, 1.22 Hz, 1H ), 4.86 (d, J = 5.14 Hz, 2H), 4.61 (q, J = 6, 93 Hz, 2 H), 2.17 (s, 3H), 1.35 (t, J = 7.09 Hz, 3H). LCMS (LCMS Method E): retention time = 0.75 min, [M + H] ⁺ = 353.2.

Stage 3:

(E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^ ] imidazole-5-carboxamide)

- 48 036435

To a solution of 1-allyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5-carboxamide (70 mg, 0.199 mmol) in DCM (1.0 ml) and MeOH (1.0 ml) was added dropwise a solution of ptoluenesulfonic acid monohydrate (37.8 mg, 0.199 mmol) in MeOH (1.0 ml) and the resulting clear solution was concentrated in vacuo. The glassy evaporation residue was stirred with DCM (4.0 ml) until a milky suspension was obtained. 2nd generation Hoveyda-Grubbs catalyst (18.67 mg, 0.030 mmol) was added to a 5 ml Biotage® sealed tube under N2. Then the above milk suspension was added and the mixture was heated to 8 0 ° C for 4 hours in a microwave reactor. At the end of this period, 5.0 ml of MeOH was added followed by a solution of 1.0 M KHMDS in THF (0.25 ml) in MeOH (1.0 ml). The mixture was stirred for 5 minutes at room temperature, concentrated in vacuo and subjected to normal phase silica gel chromatography (Biotage® Ultra SNAP silica cartridge 25 g, 0-40% MeOH / DCM gradient mixture) to give a greenish white solid. The solid was then washed with 0.2 ml of MeOH to remove the dark green ruthenium residue to give (E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3methyl -1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5-carboxamide) (14 mg, 0.02 mmol, 19.8% yield). ¹ H NMR (400 MHz, DMSO-d;) δ ppm 12.82 (br s, 2H), 7.97 (s, 2H), 7.94 (br s, 2H), 7.71 (dd , J = 8.34, 1.26 Hz, 2H), 7.44 (d, J = 8.34 Hz, 2H), 7.34 (br s, 2H), 6.55 (s, 2H) , 5.93 (br s, 2H), 4.83 (br s, 4H), 4.53 (q, J = 6, 82 Hz, 4H), 2.12 (s, 6H), 1, 27 (t, J = 7.07 Hz, 6H); LCMS (LCMS Method C): retention time = 0.79 min, [M + H] + = 677.5.

The compound obtained by the above method may be in tautomeric or isomeric form, for example, in the form (2E, 2'E) -1,1 '- ((E) -but-2-en-1,4-diyl) bis ( 2- ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl) imino) -2,3-dihydro-1H-benzoU] imidazole-5-carboxamide)

Example 3. 1,1 '- ((Methylazanediyl) bis (ethane-2,1-diyl)) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole -5-carboxamide) trifluoroacetate

- 49 036435

Stage 1:

4,4 '- (((Methylazanediyl) bis (ethane-2,1-diyl) bis (azandiyl)) bis (3-nitrobenzamide)

A mixture of% - (2-aminodhy.l ') -% - methyldga11-1,2-diamine (0.318 g, 2.72 mmol), K ₂ CO ₃ (1.501 g, 10.86 mmol) and 4-fluorine -3-nitrobenzamide (1 g, 5.43 mmol) in DMSO (20 mt) was stirred at room temperature overnight. Water was added and the resulting precipitate was collected by filtration and dried under reduced pressure to give 4,4 '- ((methitazandiit) bis (ethane-2,1-diit)) bis (azandiyl)) bis (3-nitrobenzamide) (800 mg, 1 , 62 mmol, 59.6% yield) as a yellow solid. LCMS (LC-MS Method A): retention time = 1.01 min, [M + H] + = 446.

Stage 2:

1,1 '- ((Metitazandiite) bis (ethane-2,1-diit)) bis (2-amino-1H-benzo [d] imidazole-5-carboxamide)

4,4 '- (((Methitazandiite) bis (ethane-2,1-diit)) bis (azandiite)) bis (3-nitrobenzamide) (700 mg, 1.572 mmol) and 10% Pd / C (84 mg, 0.079 mmol) in a mixture of NMP (20 ml) and MeOH (30 ml) was stirred under an atmosphere of hydrogen gas at room temperature overnight. The catalyst was removed by filtration and the MeOH was removed under reduced pressure. Cyanogen bromide (416 mg, 3.93 mmol) was then added and the reaction mixture was stirred at 60 ° C for 4 h. Et ₂ O was added and the resulting precipitate was collected by filtration and dried under reduced pressure to give 1,1 '- ((methylazandiite) bis (ethane-2,1-diitol)) bis (2-aminobenzo-1H- [d] imidazole-5-carboxamide) (500 mg, 1.03 mmol, 65.8% yield) as a red solid. LCMS (LCMS Method A): retention time = 0.94 min, [M + H] ⁺ = 435.8.

Stage 3:

1,1 '- ((Methylazanediyl) bis (ethane-2, 1-diyl)) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo ^] imidazole-5-carboxamide ) trifluoroacetate

- 50 036435

To a mixture of 1,1 '- ((methylazanediyl) bis (ethane-2,1-diyl)) bis (2-amino-1H-benzoH | imidazole-5-carboxamide) (300 mg, 0.689 mmol), 1-ethyl- DIPEA (267 mg, 2.067 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was quenched with water (30 ml) and the reaction mixture was extracted with DCM (3 x 50 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (Gilson® column, Gemini® C18, gradient 3595% MeCN: H ₂ O 0.1% TFA) to give 1,1 '- ((methylazanediyl) bis (ethane-2,1-diyl) ) bis (2- (1-ethyl-3methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide) trifluoroacetate (130 mg, 0.18 mmol, 26% yield) as a gray solid substances. Ή NMR (400 MHz, DMSO ^ 6) δ ppm 12.82 (s, 1H), 7.98 (s, 4H), 7.74 (d, J = 8.0 Hz, 2H), 7.47 ( s, 2H), 7.37 (s, 2H), 6.59 (s, 2H), 4.56 (d, J = 6.7 Hz, 4H), 4.18 (s, 4H), 3, 35 (s, 8H), 2.09 (s, 6H), 1.32-1.25 (m, 6H), LCMS (LCMS Method A): retention time = 1.14 min, [M + H] + = 708.

Example 4. Methyl 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2 - (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo [d] imidazole-5-carboxylate

- 51 036435

Stage 1:

tert-Butyl (4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) carbamate

A mixture of tert-butyl (4-aminobutyl) carbamate (5.00 g, 26.6 mmol), 4-fluoro-3-nitrobenzamide (4.89 g, 26.6 mmol) and K ₂ CO ₃ (4.04 g , 29.2 mmol) in DMSO (25 ml) was stirred at 70 ° C for 2 h. The reaction mixture was cooled to room temperature and slowly diluted with 125 ml of water through an addition funnel. The resulting solid was isolated by filtration, dried on a Buchner funnel and placed in a vacuum oven at 5b ° C for 3 days to afford the title compound (9.2 g, 26.1 mmol, 98% yield) as a yellow solid. 1H NMR (400 MHz, DMSO 6) δ ppm 8.67 (d,

- 52 036435

J = 2.02 Hz, 1H) 8.40 (t, J = 5.43 Hz, 1H), 8.01 (d, J = 6.82 Hz, 2H), 7.30 (br s, 1H ) 7.12 (d, J = 9.09 Hz, 1H), 6.87 (br s, 1H), 3.42 (q, J = 6.57 Hz, 2H), 2.91-3, 01 (m, 2H) 1.60 (d, J = 6.57 Hz, 2H) 1.43-1.54 (m, 2H) 1.38 (s,

9H). LCMS (LCMS Method C): retention time = 0.86 min, [M + H] + = 353.

Stage 2:

tert-Butyl (4 - ((2-amino-4-carbamoylphenyl) amino) butyl) carbamate

In a round-bottom flask with a capacity of 500 ml was loaded tert-butyl (4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) carbamate (9.2 g, 26.1 mmol), 10% Pd / C (0.920 g, 8 , 64 mmol) (wet, Degussa), EtOH (100 ml) and MeOH (100 ml). Air was evacuated from the flask and placed under balloon hydrogen with stirring. A refrigerator was placed on top of the flask and a hydrogen balloon was placed on top of the refrigerator. The mixture was stirred at room temperature for 20 h, then air was evacuated from the flask and the suspension was filtered through a Celite® pad using EtOH to facilitate washing. The filtrate was concentrated in vacuo and placed under high vacuum to give the title compound (8.4 g, 26.1 mmol, 100% yield) as a black solid. ¹ H NMR (400 MHz,

DMSOO δ ppm 7.44 (br s, 1H) 7.04-7.15 (m, 2H), 6.85 (t, J = 5.43 Hz, 1H), 6.74 (br s 1 H), 6.37 (d, J = 8.08 Hz, 1H), 4.89 (t, J = 5.18 Hz, 1H), 4.60 (br s, 2H), 3.07 ( q, J = 6.48 Hz, 2H), 2.97 (q, J = 6.40 Hz, 2H), 1.45-1.64 (m, 4H), 1.39 (s, 9H). LCMS (LCMS Method C): retention time = 0.68 min, [M + H] + = 323, 1.

Stage 3:

tert-Butyl (4- (2-amino-5-carbamoyl-1H-benzo-imidazol-1-yl) butyl) carbamate hydrobromide

tert-Butyl (4 - ((2-amino-4-carbamoylphenyl) amino) butyl) carbamate (8.40 g, 26.1 mmol) was dissolved in MeOH (110 ml) and a 5 M solution of cyanogen bromide in CH _{3 was} added via syringe CN (5.73 ml, 28.7 mmol). The dark reaction mixture was covered and stirred for 15 hours at room temperature. The reaction mixture was concentrated in vacuo and placed under high vacuum to give the title compound (11.17 g, 26.1 mmol, 100% yield) as a dark solid. 1H NMR (400

MHz, DMSOO δ ppm 12.85 (br s, 1H), 8.74 (br s, 2H), 8.08 (br s, 1H), 7.80-7.90 (m, 2H) 7.64 (d, J = 8.34 Hz, 1H), 7.44 (br s, 1H), 6.89 (t, J = 5.56 Hz, 1H), 4.15 (t, J = 7.20 Hz, 2H), 2.96, J = 6.32 Hz, 2H), 1.66 (d, J = 7.07 Hz, 2H), 1.42-1.50 (m, 2H ), 1.38 (s, 9H). LCMS (LCMS Method C): retention time = 0.62 min, [M + H] + = 34 8.1.

Stage 4:

tert-Butyl (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazol1-yl) butyl) carbamate

A mixture of tert-butyl (4- (2-amino-5-carbamoyl-1H-benzoIimidazol-1-yl) butyl) carbamate hydrobro- 53 036435 mid (11.17 g, 26.1 mmol), 1-ethyl-3- methyl-1H-pyrazole-5-carboxylic acid (4.82 g, 31.3 mmol), HATU (11.90 g, 31.3 mmol), DIPEA (18.22 ml, 104 mmol) and HOBt (1.997 g , 13.04 mmol) in DMF (100 ml) was stirred at room temperature for 21 h. The reaction mixture was diluted with 300 ml of water and 300 ml of EtOAc, transferred to a separatory funnel, the layers were separated and the aqueous layer was extracted with EtOAc (2 x 150 ml) ... The combined EtOAc layers were washed with saturated NH ₄ Cl (2x200 ml), water (1x200 ml) and brine (2x200 ml). The organic layer was dried over Na ₂ SO ₄ , filtered, concentrated in vacuo and placed under high vacuum. The solid was purified by silica gel chromatography (Isco® Combiflash, 0-20% MeOH: DCM, 330 g column, loaded in 50 ml DCM). The desired fractions were combined, concentrated in vacuo and placed under high vacuum to give the title compound as a purple solid (9.53 g, 19.71 mmol, 76% yield). 1H NMR (400 MHz, DMSO ^) δ ppm 12.85 (s, 1H), 8.01 (br s, 2H), 7.81 (d, J = 8.34 Hz, 1H), 7.59 (d, J = 8.34 Hz, 1 H), 7.36 (br s, 1H), 6.80-6.86 (m, 1H), 6.68 (s, 1H), 4.64 (q, J = 6, 82 Hz, 2H), 4.23 (t, J = 6.44 Hz, 2 H) 2.98 (d, J = 5.81 Hz, 2H), 2.19 (s , 3H) 1.76 (d, J = 6.57 Hz, 2H) 1.40-1.48 (m, 2H) 1.30-1.40 (m, 13H). LCMS (LCMS Method C): retention time = 0.89 min, [M + H] + = 484.3.

Stage 5:

1- (4-Aminobutyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoHimidazole-5carboxamide 2 hydrochloride

Ice-cooled 500 ml round bottom flask containing tert-butyl (4- (5carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoSimidazol-1-yl) butyl) carbamate (9.53 g, 19.71 mmol) was treated with 4 M HCl in 1,4-dioxane (42.0 ml, 168 mmol). The ice bath was removed and the purple suspension was stirred at room temperature for 2.5 h. The reaction mixture was then concentrated in vacuo, placed in a high vacuum and the resulting solid was placed in a vacuum oven at 50 ° C for 15 h and cooled in a high vacuum to give the crude title compound as a gray solid which also contained 1,4-dioxane (11.89 g, presumably 19.7 mmol, 100% yield). The substance was used as such without further purification. 1H NMR (400 MHz, DMSO-06) δ ppm 12.91 (br s, 1H), 8.03 (d, J = 1.26 Hz, 2H), 7.77-7.87 (m, 4H ), 7.62 (d, J = 8.34 Hz, 1H), 7.38 (br s, 1H), 6.70 (s, 1H), 6-5 (br s, 1H), 4 , 63 (q, J = 7.07 Hz, 2H), 4.28 (t, J = 6.57 Hz, 2H), 2.77-2.87 (m, 2H), 2.20 (s, 3H) 1.81-1.91 (m, 2H) 1.52-1.60 (m, 2H) 1.38 (t, J = 7.07 Hz, 3H). LCMS (LCMS Method C): retention time = 0.60 min, [M + H] + = 384, 2.

Stage 6:

Methyl 4 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoShimidazol-1yl) butyl) amino) -3-methoxy-5-nitrobenzoate

In a three-necked round-bottom flask with a capacity of 250, equipped with a condenser, a large stirrer and an internal thermometer, was charged 1- (4-aminobutyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -1H-benzo-thimidazole-5-carboxamide dihydrochloride (9.38 g, 20.55 mmol) and methyl 4-chloro3-methoxy-5-nitrobenzoate (5.048 g, 20.55 mmol). DMSO (50 ml) was added followed by DIPEA (17.95 ml, 103 mmol) and the dark suspension was heated at 100 ° C for about 24 h, cooled and added dropwise to 500 ml of stirring water. After the addition was complete, the resulting orange suspension was stirred for 20 minutes and filtered. The isolated orange-red paste was washed with water and hexane, dried on a Buchner funnel and then in a vacuum oven at 56 ° C for 20 hours. Then the reddish solid was triturated with Et ₂ O (60 ml) and separated by filtration. Trituration and filtration were repeated. The resulting solid was placed in a vacuum oven at 5b ° С for 3 days.

54 036435 current to give the title compound (11.17 g, 18.85 mmol, 92% yield) as a reddish solid. ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.78 (br s, 1H) 8.12 (s, 1H), 7.99 (s, 1H), 7.93 (d, J = 7 , 53 Hz, 2H) 7.79 d, J = 8.28 Hz, 1H), 7.53 (d, J = 7.78 Hz, 1H), 7.36 (s, 1H), 7.31 ( br s, 1H), 6.60 (s, 1H), 4.60 (d, J = 7.03 Hz, 2H), 4.23 (br s, 2H), 3.84 (s, 3H ), 3.80 (s, 3H) 3.53 (d, J = 5.77 Hz, 2H), 2.15 (s, 3H) 1.82 (br s, 2H), 1.62 (br s, 2H), 1.35 (t, J = 7.03 Hz, 3H). LCMS (LCMS Method D): retention time = 0.67 min, [M + H] + = 711.6.

Stage 7:

Methyl 3-amino-4 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) butyl) amine ) -5-methoxybenzoate

Methyl 4 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoShimidazol-1yl) butyl) amino) -3-methoxy-5-nitrobenzoate (5.0 g, 8.44 mmol) was mainly dissolved in DMF (50 ml) with stirring at room temperature in a 250 ml round bottom flask. Raney nickel (Raney nickel in water, about 10 ml slurry, Aldrich) was added and a condenser was attached to the flask. A three-way stopcock with an attached hydrogen cylinder was placed on top of the refrigerator, and air was evacuated from the installed system, filled with hydrogen, evacuated, and finally filled with hydrogen. The reaction mixture was heated at 70 ° C for 7 h. An additional 8 ml of Raney nickel suspension was added and the reaction mixture was heated at 70 ° C for 14 h. The reaction mixture was cooled and filtered through Celite®, washing with DMF. The filtrate, a DMF solution of about 100 ml and 20 ml of water from the Raney nickel suspension containing the desired product were used as a solution directly in the next reaction. Presumably a quantitative yield. LCMS (LCMS Method D): retention time = 0.73 min, [M + H] + = 563.4.

Stage 8:

Methyl 2-amino-1 - (4- (5-carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) - 7-methoxy-1H-benzoSimidazole-5-carboxylate hydrobromide

Methyl 3-amino-4 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) butyl) amino ) -5-methoxybenzoate (solution in DMF / water from the previous step) was treated with a 5 M solution of cyanogen bromide in CH ₃ CN (1.875 ml, 9.37 mmol) and the resulting solution was stirred at room temperature for 22 h. The reaction mixture was concentrated into vacuum and placed under high vacuum to give a brown semi-solid. The semi-solid was triturated with EtOAc, stirred vigorously for 30 min and the resulting solid was collected by filtration and dried on a Buchner funnel to give the crude title product as a tan solid (5.08 g). This crude material was used without purification. LCMS (LCMS Method D): retention time = 0.72 min, [M + H] + = 588.5.

Example 4.

Stage 9:

Methyl 1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazol-1 yl) butyl) -2- (1-ethyl-3 -methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo Schimidazole-5-carboxylate

- 55 036435

A mixture of methyl 2-amino-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoHimidazol-1-yl) butyl) -7-methoxy -1H-benzo-hydroxyimidazole-5-carboxylate hydrobromide (5.073 g, 7.59 mmol), 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (1.277 g, 8.35 mmol), HATU (3.46 g, 9.11 mmol) and DIPEA (3.98 ml, 22.76 mmol) in DMF (30 ml) were stirred at room temperature for 17 h. The reaction mixture was concentrated in vacuo, then the resulting residue was triturated with water (100 ml ) and stirred for 30 minutes. The resulting slurry was filtered and partially dried on a Buchner funnel to give a dark yellow-brown solid. The solid was mainly dissolved in 150 ml of a mixture of 10% 1PA: chloroform, diluted with water and filtered. The layers of the filtrate were then separated and the organic layer was dried over Na ₂ SO ₄ , filtered, concentrated and placed under high vacuum to give a tan solid. The solid was triturated with a warm mixture of 10% 1PA: chloroform (100 ml) and filtered. The layers of the filtrate were separated, the organic layer was dried over Na ₂ SO ₄ , filtered, added to the starting yellow-brown solid, concentrated in vacuo, and placed under high vacuum. The solid was purified by silica gel chromatography (Biotage® Isolera, 120 g Gold column, 0-10% MeOH: DCM for 30 min, applied as a solution in DCM / MeOH). The desired product fractions were combined, concentrated and placed under high vacuum to give a light yellow brown solid. The solid was triturated with DCM (50 ml) and collected by filtration and placed in a vacuum oven at 56 ° C for 30 h to give methyl 1- (4- (5-carbamoyl-2- (1-ethyl-3methyl-1H-pyrazole- 5-carboxamido) -1H-benzoSimidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -7-methoxy-1H-benzoShimidazole-5-carboxylate as a white solid (1.0 g, 1.4 mmol, 18% yield). ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.89 (s, 1H) 12.82 (s, 1H) 7.90-8.01 (m, 2H) 7.70-7.81 (m , 2H), 7.53 (d, 8.28 Hz, 1H), 7.30-7.40 (m, 2H), 6.59 (d, J = 5.02 Hz, 2H), 4.50 -4.44 (m, 4H), 4.38 (br s, 2H), 4.27 (br s, 2H), 3.87 (d, J = 3.76 Hz, 6H), 2, 10 (s, 6H), 1.86 (brs, 4H), 1.23-1.39 (m, 6H). LCMS (LC-MS Method D): retention time = 1.0-0 min, [M + H] + = 724.5.

Example 5. 1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazol 1 -yl) butyl) -2- (1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo Schimidazole-5carboxamide ditrifluoroacetate

Methyl 1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazol-1 yl) butyl) -2- (1-ethyl-3 -methyl-1Н-pyrazole-5-carboxamido) -7-methoxy-1Н-benzo-imidazole-5-carboxylate (0.1624 g, 0.224 mmol) was suspended in NH4OH (50 ml, 725 mmol) and the reaction mixture was stirred for 6 days at room temperature. The reaction mixture was concentrated in vacuo and the residue was purified by HPLC (Gilson® Autoprep, Luna acid column, loaded as a solution in DMSO, 20-50% MeCN: water with 0.1% TFA). The desired fractions were combined and concentrated to give a white solid. The solid was purified again (Gilson® Autoprep, Luna acid column, loaded as a solution in DMSO, 20-50% MeCN: water with 0.1% TFA) and the desired fractions were pooled, concentrated, placed under high vacuum and then dried in vacuum oven for 15 hours at 56 ° C to obtain 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -1H-benzo-^ imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7 methoxy-1H-benzo-imidazole-5-carboxamide ditrifluoroacetate as a white solid (76

- 56 036435 mg, 0.081 mmol, yield 36%). 1H NMR (400 MHz, DMSO-de) δ ppm 12.79 (br s, 1H) 7.97 (d, J = 1.47 Hz,

3H) 7.76 (dd, J = 8.56, 1.47 Hz, 1H) 7.64 (d, J = 1.22 Hz, 1H), 7.53 (d, J = 8.31 Hz, 1H), 7.27-7.39 (m, 3H), 6.60 (d,

J = 8.31 Hz, 2 H) 4.57 (quint, J = 7.09 Hz, 4H), 4.37 (br s, 2H), 4.28 (br s, 2H), 3, 82 (s, 3H), 2.11 (d, J = 4.16

Hz, 6H), 1.86 (br s 4H) 1.31 (td, J = 7.03, 4.52 Hz, 6H). LCMS (LCMS Method E): retention time = 0.85 min, [M + H] + = 709.5.

Example 6 (E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- ( 3hydroxypropoxy) -1H-benzoOl | imidazole-5-carboxamide) ditrifluoroacetate

Stage 1:

3- (3 - ((tert-Butyldimethylsilyl) oxy) propoxy) -4-chloro-5-nitrobenzamide

(3-Bromopropoxy) (tert-butyl) dimethylsilane (7.3 g, 28.8 mmol) was dissolved in dry DMF (75 ml), 4-chloro-3-hydroxy-5-nitrobenzamide (4.8 g, 22 , 16 mmol) and then K2CO3 (6.13 g, 4 4.3 mmol) and stirred for 2 h at 100 ° C under nitrogen atmosphere. The reaction mixture was cooled to room temperature, poured into EtOAc (600 ml), washed with water (600 ml), brine, dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with 20-80% hexane / EtOAc to give the title compound (7.43 g, 19.1 mmol, 86% yield). 1H NMR (400 MHz, DMSOO δ ppm 8.29 (br s, 1H), 8.05 (d, J = 1.71 Hz, 1H), 7.89 (d, J = 1.71 Hz, 1H ), 7.77 (br s, 1H), 4.30 (t, J = 5.99 Hz, 2H), 3.80-0.90 (m, 9H), 0.02 (s, 6H) LCMS (LC-MS Method E): retention time = 1.40 min, [M + H] + = 389.

Stage 2:

4- (Allylamino) -3- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -5-nitrobenzamide

3- (3 - ((tert-Butyldimethylsilyl) oxy) propoxy) -4-chloro-5-nitrobenzamide (2.05 g, 5.27 mmol) was dissolved in dry NMP (12 ml), allylamine (1.204 g, 21 , 08 mmol) and the reaction mixture was heated to 120 ° C in a microwave reactor for 30 min. Additional allylamine (900 mg, 15.8 mmol) was added to the reaction mixture and heated at 120 ° C for an additional 20 min. The reaction mixture was poured into EtOAc (150 ml), washed with water (150 ml), brine, dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with 20-80% hexane / EtOAc to give the title compound (1.99 g, 4.86 mmol, 92% yield). 1H NMR (400 MHz, DMSOD6) δ ppm 8.19 (s, 1H), 8.02 (br s, 1H), 7.74 (t, J = 6.0.02 Hz, 1H), 7.57 ( s, 1H), 7.31 (br s, 1H), 5.89 (ddt, J = 16.53, 10.89, 5.36, 5.36 Hz, 1H), 5.05-5, 19 (m, 2H), 4.09-4.22 (m, 4H), 3.79 (t, J = 5.90 Hz, 2H), 1.99 (t, J = 5.77 Hz, 2H ), 0.87 (s, 9H), 0.04 (s, 6H). LCMS (LCMS Method D): retention time = 1.41 min, [M + H] + = 410.

Stage 3:

4- (Allylamino) -3-amino-5- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) benzamide

- 57 036435

4- (Allylamino) -3- (3- (tert-butyldimethylsilyl) oxy) propoxy) -5-nitrobenzamide (1.91 g, 4.66 mmol) was dissolved in AcOH (13.3 ml), zinc powder (1.220 d, 18.65 mmol) (in one portion) and the reaction mixture was stirred at room temperature under nitrogen atmosphere. After 45 min, additional zinc (610 mg, 9.32 mmol) was added and stirred for an additional 2 h at room temperature. The reaction mixture was filtered, the filtrate was poured into EtOAc (125 ml), washed with 10% aqueous Na ₂ CO ₃ (125 ml), brine, dried over MgSO ₄ , filtered and concentrated in vacuo to give the title compound. 1H NMR (4000 MHz, DMSOhO δ ppm 7.60 (br s, 1H), 6.93 (d, J = 8.80 Hz, 1H), 6.85 (d, J = 1.71 Hz, 1H) (d, J = 1.96 Hz, 1H), 5.825.95 (m, 1H), 5.14 (dd, J = 17.12, 1.96 Hz, 1H), 4.95-5, 08 (m, 1H), 4.68 (br s, 1H), 3.97-4.07 (m, 2H), 3.713.86 (m, 2H), 3.60 (d, J = 5, 87 Hz, 1H), 1.84-1.96 (m, 4H), 0.75-0.92 (m, 9H), 0.02-0.08 (m, 6H). LCMS (LCMS method D): retention time = 1.04 min, [M + H] + = 380.

Stage 4:

1-Allyl-2-amino-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1H-benzoHimidazole-5-carboxamide hydrobromide

4- (Allylamino) -3-amino-5- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) benzamide (1.769 g, 4.66 mmol) was dissolved in dry MeOH (25 ml), cyanogen bromide (0.543 g, 5.13 mmol) and the reaction mixture was stirred overnight at room temperature under nitrogen atmosphere. The reaction mixture was concentrated in vacuo and the residue was stirred with EtOAc (20 ml) at room temperature for 30 min. The solids were isolated by filtration and dried to give the title compound (1.56 g, 3.21 mmol, 69% yield). 1H NMR (400 MHz, DMSOD6) δ ppm 12.94 (brs, 1H), 8.60 (brs, 2H), 8.08 (brs, 1H), 7.51 (d, J = 0.98 Hz, 1H), 7.43 (d, J = 0.98 Hz, 2H), 5.92-6.08 (m, 1H), 5.21 (dd, J = 10.51, 0.98 Hz, 1H), 4.98-5.08 (m, 1H), 4.92 (d, J = 4.65 Hz, 1H), 4.16-4.29 (m, 2H), 3.74-3.81 (m, 2H), 1.93-2.07 (m, 2H) 0.81-0.91 (m, 9H), 0.04-0.07 (m, 6H) ... LCMS (LCMS Method D): retention time = 1.02 min, [M + H] + = 405.

Stage 5:

-Alyl-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -1H-benzoShimidazole-5-carboxamide

1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (0.579 g, 3.76 mmol), HATU (1.429 g, 3.76 mmol) and HOBt (0.240 g, 1.565 mmol) were combined with dry DMF ( 12 ml). Et ₃ N (1.7 ml, 12.52 mmol) was added and the reaction mixture was stirred at room temperature for 5 min. To the reaction mixture was added 1-allyl-2-amino-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1H-benzoSimidazole 5-carboxamide hydrobromide (1.52 g, 3.13 mmol) and stirred at room temperature overnight under nitrogen. The reaction mixture was poured into EtOAc (120 ml), washed with water (120 ml), brine, dried (MgSO ₄ ), filtered and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with 80-100% EtOAc / hexane to afford the title compound (1.07 g, 1.98 mmol, 63% yield). ¹ H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.85 (br s, 1H), 7.91-8.05 (m, 1H), 7.67 (s, 1H), 7.37 (s , 1H), 7.32 (br s, 1H), 6.63 (s, 1H), 5.96-6.13 (m, 1H), 5.14 (d, J = 9.29 Hz, 1H), 4.91-5.03 (m, 3H), 4.61 (q, J = 7.01 Hz, 2H), 4.24 (t, J = 5.87 Hz, 2H), 3, 81 (t, J = 6.11 Hz, 2H), 2.18 (s, 3H) 2.07 (m, 2H), 1.34 (t, J = 7.09 Hz, 3H), 0.80 -0.92 (m, 9H), 0.04 (s, 6H). LC-MS (LC-MS

- 58 036435 method D): retention time = 1.40 min, [M + H] + = 541.

Stage 6:

-Alyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo [d] uMudazole-5-carboxamide one

-Alyl-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo | imidazole-5-carboxamide (700 mg, 1.30 mmol) was dissolved in dry THF (6 ml), AcOH (0.15 ml, 2.60 mmol) was added followed by the addition of TBAF (2.6 ml, 1M solution in THF). The reaction mixture was stirred overnight at room temperature under nitrogen atmosphere and poured into EtOAc and water (40 ml each) and shaken vigorously. The insoluble matter was filtered off and dried to give 1-allyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3hydroxypropoxy) -1H-benzoSimidazole-5-carboxamide (4 60 mg, 1 , 08 mmol, 83%). 1H NMR (400 MHz, DMSO-d) δ ppm 12.84 (br s, 1H), 7.99 (br s, 1H), 7.67 (s, 1H), 7.38 (s, 1H ) 7.32 (br s, 1H), 6.62 (s, 1H), 5.98-6.12 (m, 1H), 5.15 (d, J = 9.05 Hz, 1H), 4.92-5.04 (m, 3H), 4.54-4.68 (m, 3H), 4.24 (t, J = 6.24 Hz, 2H), 3.63 (q, J = 6.11 Hz, 2H), 2.18 (s, 3H), 1.97 (quint, J = 6.17 Hz, 2H), 1.35 (t, J = 7.09 Hz, 3H). LCMS (LCMS Method D): retention time = 0.79 min, [M + H] + = 427.

Example 6.

Stage 7:

(E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy ) -1Н-benzo (U) | imidazole-5-carboxamide) ditrifluoroacetate

-Alyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo H | imidazole-5-carboxamide (100 mg, 0.23 mmol) dissolved in DCM: MeOH 1: 1 (5 ml). To the solution was added TsOH-H ₂ O (45 mg, 0.23 mmol) as a solution in MeOH (1.5 ml) and the reaction mixture was concentrated in vacuo. DCM (5 ml) was added to the residue and the fine suspension was transferred into a microwave vial containing (1,3-dimethylimidazolidin-2-ylidene) (2-isopropoxybenzylidene) ruthenium (VI) chloride (22 mg, 0.035 mmol). The flask was degassed and heated at 80 ° C in a microwave reactor for 3 hours. The reaction mixture was treated with MeOH (3 ml) and evaporated under nitrogen. The residue was purified by HPLC (Gilson®, eluting with 10-60% ACA / water / 0.1% TFA) and the product containing fractions were collected and lyophilized to give (E) -1,1 '- (but-2 -ene-1,4-diyl) bis (2 (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo [d] imidazole-5-carboxamide) ditrifluoroacetate (57 mg, 23% yield). 1H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.84 (brs, 2H), 7.99 (brs, 2H), 7.64 (s, 2H), 7.35 (brs, 2H), 7.31 (s, 2H), 6.52 (s, 2H), 5.81 (br s, 2H), 4.91 (br, 4H), 4.52 (q, J = 6.93 Hz, 5H), 4.02 (t, J = 6.36 Hz, 5H), 3.41 (t, J = 5.99 Hz, 4H), 2.06-2.15 (m, 6H), 1.59-1.70 (m, 4H), 1.27 (t, J = 7.09 Hz, 6H). LCMS (LCMS Method D): retention time = 0.81 min, [M + H] + = 825.

The compound obtained by the above method may be in tautomeric or isomeric form, for example in the form (2E, 2'E) -1,1 '- ((E) -but-2-en-1,4-diyl) bis (2 - ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl) imino) -7- (3-hydroxypropoxy) -2,3-dihydro-1H-benzo-imidazole-5-carboxamide) ditrifluoroacetate

- 59 036435

Example 7. 8-Ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-tetradecahydrobenzo [4 , 5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15, 17] pentaazacyclogeneicosin-3,24-dicarboxamide

Stage 1:

-Alyl-2- (1- (5 - (5 - ((1-allyl-5-carbamoyl-1H-benzoI imidazol-2-yl) carbamoyl) -1-ethyl-3 -methyl1 H-pyrazol-4-yl ) pentyl) -3-methyl-1H-pyrazole-5-carboxamido) -1 H-benzo (^] imidazole-5-carboxamide

A 5.0 ml Biotage® sealed tube was loaded with 4- (5- (5-carboxy-3-methyl1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (634 mg, 1.820 mmol), 1allyl-2-amino-1H-benzo ^] imidazole-5-carboxamide hydrobromide (1352 mg, 4.55 mmol), HATU (1730 mg, 4.55 mmol) and NMP (13 ml). After stirring for 1 min at room temperature, DIPEA (3.17 ml, 18.20 mmol) was added and the mixture was stirred at room temperature for 5 min, then heated in a microwave reactor at 140 ° C for 1 h. At the end of this period, 5.0 ml of water was added and the mixture was stirred at room temperature for 5 minutes. The mixture was then poured into 2 50 ml ice water and stirred vigorously for 1 hour.The resulting solid was filtered, washed with water, washed off the filter using MeOH / DCM, concentrated in vacuo and chromatographed on silica gel (Biotage® Ultra SNAP SiO column ₂ 100 g: 0-40% MeOH / EtOAc mixture) to give 1-allyl-2- (1- (5- (5 - ((1-allyl-5-carbamoyl-1H-benzo ^] imidazole-2

- 60 036435 yl) carbamoyl) -1-ethyl-3-methyl-1H-pyrazol-4-yl) pentyl) -3-methyl-1H-pyrazole-5-carboxamido) -1HbenzoU] imidazole-5-carboxamide (840 mg , 1.128 mmol, 62% yield) as a pink solid. 1H NMR (400 MHz, DMSO-06) δ ppm 12.88 (s, 1H), 12.81 (s, 1H), 7.99-8.02 (m, 2H), 7.97 (br ( ddd, J = 8.34, 3.66, 1.39 Hz, 2H), 7.41 (dd, J = 16.93, 8.34 Hz, 2H), 7.34 (brs, 2H) , 6.65 (s, 1H) 5.87-6.02 (m, 2H), 4.99-5.22 (m, 4H), 4.82 (dd, J = 11.62, 4.80 Hz, 4H), 4.50-4.61 (m, 4H), 2.73 (t, J = 7.45 Hz, 2H), 2.15 (s, 3H), 2.08 (s, 3H ), 1.71-1.85 (m, 2H), 1.45-1.55 (m, 2H), 1.27-1.34 (m, 5H); LC-MS (LC-MS Method C ): retention time = 0.93 min, [M + H] + = 745.7.

Stage 2:

8-Ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,31-dodecahydrobenzo [4,5] imidazo [1, 2-a] benzo [4,5] imidazo [2,1-p] dipirozolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacycloneicosin-3,24 -dicarboxamide

1-allyl-2- (1 (5- (5 - ((1-allyl-5-carbamoyl- 1H-benzoI imidazol-2-yl) carbamoyl) -1 - ethyl 3-methyl- 1H-pyrazole-4yl) pentyl) -3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5-carboxamide (160 mg, 0.215 mmol), Hoveyda-Grubbs catalyst of the second generation (26.9 mg, 0.043 mmol) and only degassed 1,2-dichloroethane (DCE) (80 ml). Sealed tubes were heated in a microwave reactor for 4 h at 100 ° C. After the mixture was cooled to room temperature, MeOH (1.0 ml) was added to each tube and the resulting clear solution was stirred at room temperature for 5 minutes. A solution of potassium 2-isocyanoacetate (15 mg in 1.5 ml MeOH) was added to each tube and the resulting mixture was stirred at room temperature for 5 minutes. The contents of the tubes were combined, concentrated in a vacuum, then the residue was evaporated in a minimum volume of a DCM / MeOH mixture and purified by chromatography on silica gel (Biotage® Ultra SNAP SiO ₂ column 100 g, 0-40% MeOH / EtOAc mixture) to obtain the target product (61 mg ) as a pale green solid with a mixture of alkene isomers. The product was further purified (Biotage® Ultra SNAP SiO ₂ column 25 g, gradient 0-20% MeOH / DCM) to give 8-ethyl-10,18-dimethyl-7,20-dioxo 6,7,8,11,12, 13,14,15,20,21,28,31-dodecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1е: 4 ' , 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-3,24-dicarboxamide as a 7: 1 mixture of trans and cis isomers (54 mg, 0.075 mmol, 35% yield). Characterization of the trans isomer: ¹ H NMR (400 MHz, DMSO ^) δ ppm 12.87 (s, 1H), 12.84 (s, 1H), 7.98 (br, 4H), 7.77 (dd , J = 7.71, 3.16 Hz, 2H), 7.33-7.48 (m, 4H), 6.55 (s, 1H), 5.89-5.98 (m, 1H), 5.66-5.75 (m, 1H) d, J = 7.83 Hz, 4H), 4.73 (t, J = 6.95 Hz, 2H), 4.47 (q, J = 6, 99 Hz, 2H), 2.72-2.80 (m, 2H), 3H), 2.10 (s, 3H), 1.72 (br s, 2H), 1.44 (br s, 2H) 1.30 (t, J = 7.07 Hz, 5H); LCMS (LCMS Method C): retention time = 0.82 min, [M + H] + = 717.6.

Example 7.

Stage 3:

8-Ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-3,2 4 dicarboxamide

- 61 036435

A round bottom flask was charged with 10% Pd / C (200 mg, 0.188 mmol) and purged with nitrogen. A solution of 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31-dodecahydrobenzo was added [4, 5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17 ] pentaazacyclogeneicosine-3,24-dicarboxamide (100 mg, 0.140 mmol, 7: 1 mixture of trans and cis isomers) in a mixture of MeOH (20.0 ml) and THF (20.0 ml), the flask was purged with hydrogen and the reaction mixture stirred in a hydrogen atmosphere (1 atm) for 23 h. Then the flask was opened to air, stirred vigorously for 15 min and filtered, the Pd / C was washed with MeOH / TPA, the filtrate was concentrated in vacuo and chromatographed on silica gel (Biotage® column Ultra SNAP SiO ₂ 25 g, 0-20% MeOH / DCM mixture) to give 8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15, 20,21,28,29,30,31tetrahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3 '-1] [1,3,6,15,17] pentaazacyclogenaicosin-3,24-dicarboxamide (56 mg, 0.078 mmol, 55.8% yield) as a pale pink solid. ¹ H NMR (400 MHz, DMSO-a6) δ ppm 12.88 (brs, 2H), 8.02 (s, 4H), 7.79-7.87 (m, 2H), 7.67 ( d, J = 8.34 Hz, 1H), 7.63 (d, J = 8.34 Hz, 1H), 7.37 (br s, 2H), 6.57 (s, 1H), 4, 74 (t, J = 6.57 Hz, 2H), 4.48 (m, 2H), 2.16 (s, 3H), 2.08 (s, 3H), 1.91 (s, (m, 2H), 1.44-1.54 (m, 2H), 1.35-1.42 (m, 2H), 1.29 (t, J = 7.07 Hz, 3H)); LCMS (LCMS Method C): retention time = 0.81 min, [M + H] + 719.7.

Example 8 8-Ethyl-10,18,30-trimethyl-7,20-dioxo-7,8,11,12,13,14,15,20,21,28,29,30,31,32-tetradecahydro -1Н-benzo [4,5] imidazo [2,1-b] benzo [4,5] imidazo [1,2-i] dipyrazolo [5,1-m: 4 ', 3'-t] [1, 3,6,9,11,14] hexaazacyclodocosin-3,2 4-dicarboxamide

Example 8 8-Ethyl-10,18,30-trimethyl-7,20-dioxo-7,8,11,12,13,14,15,20,21,28,29,30,31,32-tetradecahydro -1Н-benzo [4,5] imidazo [2,1-b] benzo [4,5] imidazo [1,2-i] dipyrazolo [5,1-m: 4 ', 3'-t] [1, 3,6,9,11,14] hexaazacyclodocosin-3,24-dicarboxamide

To a solution of hexafluorophosphate 2- (7-aza-1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium (576 mg, 1.516 mmol), 1,1 '- ((methylazandiyl) bis (ethane-2 , 1-diyl)) bis (2-amino-1H-benzo ^] imidazole-5-carboxamide) (300 mg, 0.689 mmol, from example 3) and 4- (5- (5-carboxy-3-methyl-1H -pyrazol-1-yl) pentyl) -1-ethyl3-methyl-1H-pyrazole-5-carboxylic acid (240 mg, 0.689 mmol) in NMP (10 ml) was added DIPEA (267 mg, 2.067 mmol). The reaction mixture was stirred at room temperature for 0.5 h and then the reaction mixture was heated in a microwave reactor at 140 ° C for 1 h (150 W). The reaction mixture was allowed to cool to room temperature, water was added and the mixture was extracted with DCM. The organic phase was washed with water (2x20 ml), dried over anhydrous Na ₂ SO ₄ , filtered, and

- 62 036435 centered under reduced pressure. The residue was purified by preparative HPLC (Gilson® column, Gemini® C18, gradient 2-95% MeCN: H2O 0.1% TFA) to give 8-ethyl-10,18,30-trimethyl-7,20 dioxo-7,8,11 , 12,13,14,15,20,21,28,29,30,31,32-tetradecahydrobenzo [4,5] imidazo [2,1-b] benzo [4,5] imidazo [1,2-i ] dipyrazolo [5,1-m: 4 ', 3'-t] [1,3,6,9,11,14] hexaazacyclodocosine-3,24-dicarboxamide (25 mg, 0.03 mmol, 4.56% ) as a brown solid. 1H NMR (400 MHz, CD3OD) δ ppm 8.00 (d, J = 12.0 Hz, 2H), 7.92-7.87 (m, 2H), 7.57 (d, J = 8.0 Hz, 2H), 6.76 (s, 1H), 4.77-4.77 (m, 2H), 4.70-4.67 (m, 2H), 4.57-4.51 (m, 4H), 3.98 (s, 2H), 3.78 (s, 2H), 3.15 (s, 3H), 2.78 (t, J = 8.0 Hz, 2H), 2.18- 2.15 (m, 6H), 1.81-1.74 (m, 2H), 1.39-1.33 (m, 6H), 1.17-1.07 (s, 2H). LCMS (LCMS Method A): retention time = 1.26 min, [M + H] + = 748.

Example 9 1,15-bis (1-Ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -8,9,16,17,18,19-hexahydro-7Н6,10-dioxa-2,14 , 15а, 19а-tetrazacyclopentadeca [1,2,3-cd: 11,10,9-c'd '] diindene-4,12-dicarboxamide

Stage 1:

5,5 '- (Propane-1,3-diylbis (oxy)) bis (4-chloro-3-nitrobenzamide)

4-Chloro-3-hydroxy-5-nitrobenzamide (2 g, 9.23 mmol), 1,3-dibromopropane (932 mg, 4.62 mmol), DIPEA (3.23 ml, 18.47 mmol) were stirred in NMP (5 ml) in a 20 ml microwave vial. The reaction mixture was then heated in a microwave reactor at 100 ° C for 15 minutes. Water (30 ml) was added to the reaction mixture and the precipitate was collected by filtration and washed with water. The solid was then dried in vacuo at 55 ° C overnight to give the title compound (3 g, 5.71 mmol, 61.8% yield). LC-MS (LC-MS Method D): retention time = 0.99

- 63 036 435 min, [M + H] ⁺ = 473.1.

Stage 2:

1,13-Dinimro-7,8,14,15,16,17,18,19-octahydro-6H-dibenzo [b, j] [1,12,4,9] dioxadiazacyc pentadecin-3,11-dicarboxamide lo -

5,5 '- (Propane-1,3-diylbis (oxy)) bis (4-chloro-3-nitrobenzamide) (2700 mg, 5.71 mmol), butane-1, 4 diamine (503 mg, 5.71 mmol ), DIPEA (2.491 ml, 14.26 mmol) was stirred in DMSO (8 ml) in a 20 ml microwave vial. Then it was heated in a microwave oven at 120 ° C for 15 minutes. The reaction mixture was diluted with water and filtered to give the title product as an orange solid. The solid was then dried in vacuo at 55 ° C overnight to give an orange solid (ca. 2.5 g, 5.12 mmol, 90% yield). ¹ H NMR (400 MHz, DMSO <) δ ppm 8.20-8.16 (m, 2H), 8.05-7.95 (m, 4H), 7.62 (d, J = 1.71 Hz , 2H), 4.32 (t, J = 5.26 Hz, 4H), 3.56-3.42 (m, 4H), 2.47-2.38 (m, 2H), 1.65 ( br s, 4H). LCMS (LCMS Method D): retention time = 0.93 min, [M + H] + = 489.2.

Stage 3:

1,13-Damino-7,8,14,15,16,17,18,19-octahydro-bN-dibenzo [b, j] [1,12,4,9] dioxadiazacyclopentadecin-3,11-dicarboxamide

NH ₂ | о = / ^NH2 h ₂ n nh nh ₂

In a 50 ml round-bottomed flask 1,13-dinitro-7,8,14,15,16,17,18,19-octahydro-6Hdibenzo [b, j] [1,12,4,9] dioxadiazacyclopentadecin-3,11 β-dicarboxamide (2.5 g, 5.12 mmol) was stirred in AcOH (14.7 ml, 256 mmol). Then zinc (1.67 g, 25.6 mmol) was added in portions and the reaction mixture was stirred at room temperature for 1 hour. Then the reaction mixture was filtered and the filtrate was concentrated in vacuo. The material was purified by HPLC (Gilson® column, Gemini®: CH3CN, gradient 0.1% NH4OH / water 2-20%) to give the title compound (90 mg, 0.2 mmol, 3.9% yield). LCMS (LCMS Method D): retention time = 0.39 min, [M + H] + = 429.3.

Stage 4:

1,15-Diamino-8,9,16,17,18,19-hexahydro-7H-6,10-dioxa-2,14,15а, 19а-tetraazacyclopentadeca [1,2,3-cd: 11,10, 9-c'd '] diindene-4,12-dicarboxamide

In a 20 ml reaction vial, 1,13-dinitro-7,8,14,15,16,17,18,19-octahydro-6Нdibenzo ^] [1,12,4,9] dioxadiazacyclopentadecin-3,11-dicarboxamide ( 140 mg, 0.377 mmol) was stirred in MeOH (5 ml) and treated with cyanogen bromide (69.2 mg, 0.653 mmol). The reaction mixture was stirred at room temperature overnight. Then water (10 ml) was added to the reaction mixture, the solid was isolated by filtration and dried in vacuo overnight to give the title compound (100 mg, 0.188 mmol, 57.6% yield). 1H NMR (400 MHz, methanol d4) δ ppm 7.04 (d, J = 2.01 Hz, 2H), 6.97 (d, J = 1.76 Hz, 2H), 4.30 (s, 4H) , 3.04 (br s, 4H), 2.46-2.33 (m, 2H), 1.45 (br s, 4H). LCMS (LCMS Method D): retention time = 0.47 min, [M + H] + = 479.3.

Example 9 /

Stage 5:

1,15 -bis (1-Ethyl-3-methyl-1Н-pyrazole-5-carboxamido) -8,9,16,17,18,19-hexahydro-7Н-6,10-dioxa 2.14,15а, 19а -tetraazacyclopentadeca [1,2,3-cd: 11,10,9-cU] diindene-4,12-dicarboxamide

- 64 036435

To a 20 ml reaction vial was added 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (90 mg, 0.581 mmol), hexafluorophosphate 2- (3H- [1,2,3] triazolo [4,5- B] pyridin-3-yl) -1,1,3,3-tetramethylisouronium (V) (221 mg, 0.581 mmol), DIREA (0.135 ml, 0.775 mmol), DMF (5 ml) followed by the addition of 1.15- diamino-8,9,16,17,18,19-hexahydro-7Н-6,10-dioxa-2,14,15а, 19а-tetraazacyclopentadeca [l, 2,3-cd: U, 10,9-c ' d '] diindene-4,12-dicarboxamide (90 mg, 0.194 mmol). The reaction vial was sealed and heated to 140 ° C for 30 minutes. Water (20 ml) was added to the solution, and the resulting solid was collected by filtration and air dried to give a brown solid. Then the crude product was dissolved in DMSO (6 ml) and water (20 ml) was added. The resulting solid was isolated by filtration and dried in vacuo to give 1,15-bis (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -8,9,16,17,18,19-hexahydro-7H- 6,10-dioxa2,14,15а, 19а-tetraazacyclopentadeca [1,2,3-cd: 11,10,9-c'd '] diindene-4,12-dicarboxamide (50 mg, 0.063 mmol, yield 32, 7%) as a light brown solid. ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.84 (br s, 2H), 8.00 (br s, 2H), 7.67 (s, 2H), 7.46 (s, 2H) 7.37 (br s, 2H), 6.59 (s, 2H), 4.60 (d, J = 6.78 Hz, 4H), 4.48 (d, J = 4.52 Hz , 4H), 4.38 (br s, 4H), 2.55 (s, 6H), 2.12 (s, 4H), 2.06 (d, J = 6.02 Hz, 2H), 1 , 33 (t, J = 7.03 Hz, 6H) LC-MS (LC-MS Method D): retention time = 0.92 min, [M + H] + = 751.5.

Example 10 (E) -1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) but -2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1Hbenzo [d] imidazole-5-carboxamide

- 65 036435

Stage 1:

(E) -1- (4 - ((2- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4-carbamoyl-6-nitrophenyl) amino) but-2en-1-yl) -2- (1 -ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide

In a tube for a microwave reactor containing (E) -1- (4-aminobut-2-en-1-yl) -2- (1-ethyl-3methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide hydrochloride (517 mg, 1.24 mmol, in DMSO (10 ml), added TEA (0.28 ml, 2.0 mmol) followed by K ₂ CO ₃ (274 mg, 1.98 mmol) and 3- (3 ((tert-butyldimethylsilyl) oxy) propoxy) -4-chloro-5-nitrobenzamide (385 mg, 0.990 mmol). The reaction mixture was heated to 75 ° C. After 7 h, the mixture was concentrated and the residue was purified on silica gel, eluting with 10-90% EtOAc to remove impurities, followed by elution with 0-10% MeOH in DCM to give the title compound (200 mg, 0.273 mmol, 28% yield) as an orange solid. ¹ H NMR (400 MHz, DMSO ^), 1.6 (d, J = 8.11 Hz, 2H), 7.50 (s, 1H), 7.31-7.43 (d, J = 1.52 Hz, 1H) , 7.94-8.08 (m, 3H) (m, 3H), 6.62 (s, 1H), 5.74-5.81 (m, 2H), 4.80 (brs, 2H ), 4.59 (d, J = 6.84 Hz, 2H), 4.13 (br s, 2H), 4.01 (t, J = 6.08 Hz, 2H), 3.63 (t , J = 5.96 Hz, 2H), 2.16 (s, 3H), 1.76-1.88 (m, 2H), 1.33 (t, J = 7.1 0 Hz, 3H), 0.74-0.82 (m, 9H), -0.06 (s, 6H); LCMS (LCMS Method D): retention time = 1.23 min, [M + H] + = 734.6.

- 66 036435

Stage 2:

(E) -1- (4 - ((2-amino-6- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4-carbamoylphenyl) amino) but-2en-1-yl) -2- (1 -ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo (^] imidazole-5-carboxamide

(E) -1- (4 - ((2- (3 - ((tert-Butyldimethylsilyl) oxy) propoxy) -4-carbamoyl-6-nitrophenyl) amino) but-2en-1-yl) -2- (1 -ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide (1 g, 1.363 mmol) was suspended in MeOH (20 ml) and ammonium hydroxide (4.62 ml, 34.1 mmol) and stirred for 5 min at room temperature. Then added sodium salt of hydrosulfurous acid (1.675 g, 8.18 mmol) in water (5 ml). After 60 min EtOAc (300 ml) was added and the mixture was extracted with water (50 ml x 3). The organic phase was separated, dried over Na ₂ SO ₄ and concentrated in vacuo to give the title compound (710 mg, 1.009 mmol, 74.0% yield) as a light yellow solid that was used without further purification. ¹ H NMR (400 MHz, DMSO-06) δ ppm 12.80 (brs, 1H), 8.00 (s, 1H), 7.97 (brs, 1H), 7.75 (dd, J = 8.49, 1.14 Hz, 1H), 7.63 (br s, 1H), 7.28-7.41 (m, 2H), 7.00 (br s, 1H), 6 , 84 (d, J = 1.52 Hz, 1H), 6.74 (d, J = 1.52 Hz, 1H), 6.65 (s, 1H), 5.79-5.96 (m, 1H), 5.64-5.78 (m, 1H), 4.81 (d, J = 4.82 Hz, 2H), 4.68 (br s, 2H), 4.61 (d, J = 7.10 Hz, 2H), 3.92 (t, J = 5.83 Hz, 2H), 3.84 (br s 1H), 3.63 (t, J = 6.08 Hz, 2H) , 3.57 (br s, 2H), 2.17 (s, 3H), 1.70-1.82 (m, 2H), 1.34 (t, J = 7.10 Hz, 3H), 0.68-0.83 (m, 9H); -0.06 (s, 6H); LCMS (LCMS Method J): retention time = 1.05 min, [M + H] + = 704.3.

Stage 3:

(E) -2-amino-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl1 H-pyrazole-5- carboxamido) -1 N-benzo ^] imidazole-1-yl) but-2-en-1 -yl) -1 N-benzo ^] imidazole-5 carboxamide

To a solution of (E) -1- (4 - ((2-amino-6- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4-carbamoylphenyl) amino) but-2-en-1-yl) - 2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide (120 mg, 0.177 mmol) in MeOH (5 ml) cyanogen bromide (36 mg, 0 , 34 mmol) at room temperature. After 2 h, the reaction mixture was concentrated and EtOAc (10 ml) was added. After stirring for 30 min, the solid was isolated by filtration and washed with EtOAc to give the title compound (120 mg, 0.165 mmol, 97% yield) as a light brown solid, which was used without further purification. 1H NMR (400 MHz, MeOH-d4) δ ppm 8.00 (d, J = 1.27 Hz, 1H), 7.81 (dd, J = 8.36, 1.77 Hz, 1H), 7, 49 (d, J = 1.27 Hz, 1 H), 7.39-7.45 (m, 1H), 7.36 (d, J = 1.27 Hz, 1H), 6.61 (s, 1H), 5.82-5.99 (m, 2H), 4.96-5.01 (m, 2H), 4.56-4.65 (m, 2H), 4.12 (t, J = 6.21 Hz, 2H), 3.62-3.75 (m, 2H), 2.18-2.29 (m, 3H), 1.79 (t, J = 6.21 Hz, 2H), 1.24-1.54 (m, 5H), 0.84-0.98 (m, 9H), -0.01-0.11 (m, 6H); LCMS (LCMS Method D): retention time = 0.97 min, [M + H] ⁺ = 729.5.

Stage 4:

(E) -7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) -1 H -benzo ^] imidazol-1-yl) but-2-en-1 -yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5 carboxamido) -1H-benzo ^] imidazole-5- carboxamide

- 67 036435

To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (33 mg, 0.21 mmol) in DMF (3 ml) were added HATU (75 mg, 0.20 mmol) and HOBt (12.6 mg, 0.082 mmol). After stirring at room temperature for 10 min, triethylamine (0.09 ml, 0.66 mmol) was added followed by the addition of (E) -2-amino-7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1 - (4- (5-carbamoyl-2- (1-ethyl-3methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -1H -benzo (d)] imidazole-5carboxamide (120 mg, 0.165 mmol) and the reaction was continued at room temperature. After 3 days, a solid was precipitated from the reaction mixture by the dropwise addition of water. The solid was isolated by filtration and washed with water. The solid was then purified on silica gel (Gold HP 12 g column) eluting with 0-20% MeOH in DCM. The desired fractions were combined and concentrated to give the title compound (29 mg, 0.034 mmol, 20% yield) as an off-white solid. ¹ H NMR (400 MHz, THF-d ^ δ ppm 12.53 (br s, 2H), 8.00 (d, J = 1.01 Hz, 1H), 7.61 (d, J = 1, 01 Hz, 1H) (dd, J = 8.36, 1.52 Hz, 1H), 7.36 (d, J = 6, 84 Hz, 2H), 7.29 (d, J = 1.01 Hz , 1H), 7.12 (d, J = 8.36 Hz, 1 H), 6.83 (br s, 2H), 6.66 (d, J = 2.28 Hz, 2H), 6, 06 (dt, J = 15.46, 5.58 Hz, 1H), 5.87 (dt, J = 15.46, 5.83 Hz, 1H), 5.09 (d, J = 5.32 Hz , 2H), 4.89 (d, J = 5.58 Hz, 2H), 4.59-4.72 (m, 4H), 3.97 (t, J = 6.21 (M, 2H), 1.40 (td, J = 7.03, 1.14 Hz, 6H), 0.82-0.94 (m, 9H), -0.03-0.09 (m, 6H); LC-MS (LCMS method D): retention time = 1.21 min, [M / 2 + H] + = 433.6.

Stage 5:

(E) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) - 1H-benzo ^] imidazol-1 yl) but-2-ene -1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole-5-carboxamide

To a solution of (E) -7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -1 - (4- (5-carbamoyl-2- (1-ethyl-3 methyl-1H-pyrazole-5-carboxamido) -1Н-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) -1Н-benzo ^] imidazole-5- carboxamide (25 mg, 0.029 mmol) and 1 M TBAF in THF (0.058 ml, 0.058 mmol) in THF (2 ml), acetic acid (3.3 μl, 0.058 mmol) was added at room temperature. After 12 h, the reaction mixture was concentrated, triturated with diethyl ether and EtOAc and purified on silica gel (Gold column 12 g) eluting with 0-25% methanol in DCM. The desired fractions were concentrated to afford the title compound (7 mg, 9 μmol, 32% yield) as an off-white solid. 1H NMR (400 MHz, THF-d ^ δ ppm 12.51 (br s, 2H), 8.01 (d, J = 1.01 Hz, 2H), 7.55-7.65 (m, 3H ), 7.33 (d, J = 1.01 Hz, 2H), 7.14-7.20 (m, 2H), 6.00-6.15 (m, 2H), 5.82-5, 96 (m, 2H), 5.055.13 (m, 4H), 4.04 (t, J = 6.59 Hz, 4H), 3.78-3.90 (m, 5H), 2.19 (d , J = 2.03 Hz, 6H), 1.87-2.00 (m, 2H), 1.36-1.44 (m, 6H); LC-MS (LC-MS method D): retention time = 0.79 min, [M + H] + = 751.4.

The compound obtained by the above method may be in tautomeric or isomeric form, for example in the form (E) -1 - ((E) -4 - ((E) -5-carbamoyl-2 - ((1-ethyl-3-methyl -1Н-pyrazole-5carbonyl) imino) -2,3-dihydro-1 N-benzo (^] imidazol-1-yl) but-2-en-1 -yl) -2 - ((1-ethyl-3 -methyl -1 H-pyrazole 5 -carbonyl) imine) -7- (3-hydroxypropoxy) -2,3-dihydro-1 H-benzo [d] imidazole-5-carboxamide

- 68 036435

or ^) - 1 - ((E) -4 - (^) - 5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -2,3-dihydro1H- benzo ^] imidazol-1-yl) -but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -7- (3-hydroxypropoxy ) -2,3-dihydro-III-benzo ^] imidazole-5-carboxamide

Example 11 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3hydroxypropoxy) -1 H-benzo ^] imidazole -1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-III-benzo ^] imidazole-5-carboxamide

- 69 036435

Stage 1:

(E) -tert-butyl (4 - ((4-carbamoyl-2-methoxy-6-nitrophenyl) amino) but-2-en-1-yl) carbamate

To a suspension of 4-chloro-3-methoxy-5-nitrobenzamide (1.50 g, 6.50 mmol) in EtOH (25 ml) was added (E) tert-butyl (4-aminobut-2-en-1-yl) carbamate (1.454 g, 7.81 mmol) and DIEA (3.4 ml, 20 mmol). The reaction mixture was stirred at 120 ° C in a sealed tube overnight and allowed to cool to room temperature. The resulting orange solid was collected by filtration and washed with EtOH to give the title compound (2.10 g, 5.52 mmol, 85% yield). ¹ H NMR (400 MHz, DMSO-de) δ ppm 8.19 (d, J = 1.77 Hz, 1H), 8.03 (br s, 1H), 7.76 (t, J = 6, 08 Hz, 1H) J = 1.52 Hz, 1H), 7.34 (br s, 1H), 6.95 (t, J = 5.45 Hz, 1H), 5.53 (br s, 2H), 4.09 (br s, 2H) 3.88 (s, 3H), 3.48 (br s, 2H), 1.35 (s, 9H); LC-MS (LC-MS Method D): retention time = 0.8 9 min, [Mt-Bu + H] + = 325, 1.

Stage 2:

(E) -4 - ((4-aminobut-2-en-1-yl) amino) -3-methoxy-5-nitrobenzamide hydrochloride

To a suspension of tert-butyl (E) - (4 - ((4-carbamoyl-2-methoxy-6-nitrophenyl) amino) but-2-en-1yl) carbamate (20 g, 47.3 mmol) in methanol (50 ml) was slowly added 4 M HCl in dioxane (100 ml, 400 mmol). The reaction mixture was stirred at room temperature for 1 hour, then the resulting

70 036435 the solid was isolated by filtration, washed 3 times with Et2O (100 ml x 3) and dried under high vacuum to give the title compound (13.90 g, 43.9 mmol, 93% yield). ¹ H NMR (400

MHz, DMSO ^) δ ppm 8.22 (d, J = 2.03 Hz, 1H), 7.76-8.16 (br s, 5H), 7.60 (d, J = 2.03 Hz , 1H), 7.37 (br. S,

1H), 5.87 (dt, J = 15.52, 5.80 Hz, 1H), 5.62 (dt, J = 15, 65, 6.37 Hz, 1H), 4.18 (d, J = 5.32 Hz, 2H), 3.90 (s, 3H),

3.40 (t, J = 5.70 Hz, 2H); LCMS (LCMS Method K): retention time = 0.41 min, [M + H] + = 281.1.

Stage 3:

(E) -3- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4 - ((4 - ((4-carbamoyl-2-methoxy-6-nitrophenyl) amino) but-2-ene-1 - yl) amino) -5-nitrobenzamide)

To a suspension of (E) -4 - ((4-aminobut-2-en-1-yl) amino) -3-methoxy-5-nitrobenzamide hydrochloride (9.77 g, 30.9 mmol) in 1-butanol (90 ml) sodium bicarbonate (5.18 g, 61.7 mmol) and DIEA (22.45 ml, 129 mmol) were added. The mixture was stirred at room temperature for 10 min, then 3 (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4-chloro-5-nitrobenzamide (10 g, 25.7 mmol) was added and the reaction mixture was stirred at 120 ° From during the night. The solution was allowed to cool to room temperature and the resulting dark orange solid was isolated by filtration and washed with EtOH (15 mL). Then the crude material was stirred in water (100 μl) for 10 min, filtered and washed again with water (100 ml), EtOAc (50 ml) and EtOH (20 ml). The material was dried in a vacuum oven to give the title compound (10 g, 14.54 mmol, 56.5% yield). ¹ H NMR (400 MHz, DMSO ^) δ ppm 8.16 (t, J = 1.77 Hz, 2H), 8.04 (brs, 2H), 7.72 (d, J = 5.83 Hz, 2H) (s, 2H), 7.35 (br s, 2H), 5.53-5.68 (m, 2H), 3.99-4.16 (m, 6H), 3.74 (t, J = 6.08 Hz, 2H), 3.43 (br s, 3H), 1.92 (t, J = 6.08 Hz, 2H), 0.740.88 (m, 9H), 0 .00 (s, 6H); LCMS (LCMS Method K): retention time = 1.32 min, [M + H] ⁺ = 633.4.

Stage 4:

(E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6-methoxyphenyl) amino) but-2-en-1-yl) amino) -5- (3 - (( tert-butyldimethylsilyl) oxy) propoxy) benzamide

To a solution of (E) -3- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -4 - ((4 - ((4-carbamoyl-2-methoxy-6nitrophenyl) amino) but-2-ene-1- yl) amino) -5-nitrobenzamide (5 g, 7.90 mmol) in methanol (120 ml) at 0 ° С sodium salt of hydrosulfurous acid (16.19 g, 79 mmol) in water (50 ml) and ammonium hydroxide were added (25.6 ml, 198 mmol). The reaction mixture was allowed to warm to room temperature. After 10 min at room temperature, the mixture was extracted with EtOAc (100 ml), dried over Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by silica gel chromatography (Isco column) eluting with hexane: (EtOH: EtOAc 3: 1) with the addition of 2% NH ₄ OH (0-100% gradient) to afford the title compound (2.1 g, 3 , 34 mmol, yield 42.2%). ¹ H NMR (400 MHz, DMSO-d ^ δ ppm 7.63 (br s, 2H), 6.99 (d, J = 5.58 Hz, 2H), 6.72-6.91 (m, 6H), 5.62-5.73 m, 2H), 4.66 (d, J = 8.36 Hz, 4H), 4.00 (t, J = 5.96 Hz, 2H), 3.69 -3.84 (m, 4H), 3.40-3.49 (m, 2H), 3.35 (s, 3H), 1.90 (t, J = 6.08 Hz, 2H), 0, 79-0.91 (m, 9H), 0.03-0.07 (m, 6H); LCMS (LCMS Method K): retention time = 0.4 6 min, [M + H] ⁺ = 573.3.

Stage 5:

(E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1Hbenzo U | imidazol-1 -yl ) but-2-en-1-yl) -2- (1-ethyl-3 -methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1NbenzoSimidazole-5-carboxamide

- 71 036435

To a solution of (E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6-methoxyphenyl) amino) but-2-en-1 -yl) amino) 5- (3- ( (tert-butyldimethylsilyl) oxy) propoxy) benzamide (1.02 g, 1.78 mmol) in MeOH (15 mL) was added cyanogen bromide (943 mg, 8.90 mmol). After stirring at room temperature for 20 min, a light yellow solid precipitated which was collected by filtration, washed with EtOAc and determined by LC-MS as a mixture containing about 2/3 TBDMS-protected compound (E) -2 amino -1 - (4- (2-amino-5-carbamoyl-7-methoxy-1 H-benzo ^] imidazol-1-yl) but-2-en-1-yl) 7- (3 - ((tert- butyldimethylsilyl) oxy) propoxy) -1H-benzo ^] imidazole-5-carboxamide and about 1/3 alcohol (E) -2-amino-1- (4- (2-amino-5-carbamoyl-7- (3- hydroxypropoxy) -1H-benzo (^] imidazole-1yl) but-2-en-1-yl) -7-methoxy-1H-benzo (^] imidazole-5-carboxamide, deprotected. This mixture (about 90 0 ml) was added after TEA (1.07 ml, 7.7 mmol) to a solution of 1-ethyl-3-methyl-1H-pyrazole-5 carboxylic acid (0.89 g, 5.78 mmol), HATU (2.2 g, 5.7 8 mmol) and HOBt (443 mg, 2.8 9 mmol) in DMF (10 ml), the mixture was stirred for 15 min at room temperature. After 20 h, a 5N aqueous NaOH solution (3 ml) was added. After 30 min at room temperature, water (30 mL) was added and the resulting white precipitate was collected by filtration and purified on silica gel (40 g Isco column) eluting with 0-30% MeOH in DCM to give the title compound (545 mg, 0.664 mmol , yield 38% for 2 stages). ¹ H NMR (400 MHz, DMSO-06) δ ppm 12.83 (br s, 2H), 7.99 (br s, 2H), 7.64 (d, J = 3.04 Hz, 2H) , 7.28-7.42 (m, 4H), 6.52 (s, 2H), 5.84 (bs, 2H), 4.91 (bs, 4H), 4.53 (d , J = 6.34 Hz, 4H) 4.06 (t, J = 6.34 Hz, 2H), 3.75 (s, 3H), 3.45 (t, J = 5, 96 Hz, 2H) , 2.10 (d, J = 2.53 Hz, 6H), J = 6.08 Hz, 2H), 1.27 (td, J = 7.03, 1.90 Hz, 6H); LCMS (LCMS Method D): retention time = 0.85 min, [M / 2 + H] + = 391.3833.

The compound obtained by the above method may be in tautomeric or isomeric form, for example in the form (E) -1 - ((E) -4 - ((E) -5-carbamoyl-2 - ((1-ethyl-3-methyl -1Н-pyrazol-5-carbonyl) imino) -7- (3-hydroxypropoxy) -2,3-dihydro-1Н-benzo (^] imidazol-1-yl) but-2-en-1-yl) -2- ((1-ethyl-3methyl-1H-pyrazole-5-carbonyl) imino) -7-methoxy-2,3-dihydro-1H-benzo ^] imidazole-5-carboxamide

or (7) -1 - ((E) -4 - ((7) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -7- (3 -hydroxypropoxy) -2,3-dihydro-1 H-benzo ^] imidazol-1 -yl) but-2-en-1 -yl) -2 - ((1-ethyl-3-methyl-1 H-pyrazole- 5carbonyl) imine) -7-methoxy-2,3-dihydro-1 H-benzo ^] imidazole-5-carboxamide

- 72 036435

Example 12 (E) -1,1 '- (But-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- ( 3morpholinopropoxy) -1H-benzo (^] imidazole-5-carboxamide)

HATU, HOBt.

TEA, DMF

Stage 1:

4-Chloro-3- (3-morpholinopropoxy) -5-nitrobenzamide

To a suspension of 4-chloro-3-hydroxy-5-nitrobenzamide (1.00 g, 4.62 mmol), 3-morpholinopropan-1-ol (1.00 g, 6.89 mmol) and triphenylphosphine (1.82 g , 6.93 mmol) in DCM (46 ml) was added DIAB (1.35 ml, 6.93 mmol). After stirring for 1 h, additional triphenylphosphine (480 mg, 1.83 mmol) was added and after an additional 30 min, DIEA (0.40 mL, 2.1 mmol) was added. After 1 hour, the reaction mixture was partitioned between saturated aqueous ammonium chloride and DCM. The organic layer was dried and purified on silica gel (ISCO-Rf 4 g column), eluting with 0-100% (3: 1

- 73 036435

EtOAc: EtOH) in hexane to give the title compound (630 mg, 1.83 mmol, yield

40%). 1H NMR (400 MHz, DMSO-d;) δ ppm 8.30 (s, 1H), 8.05 (d, J = 1.77 Hz, 1H), 7.88 (d, J = 1.77 Hz , 1H),

7.80 (s, 1 H), 4.28 (t, J = 6.21 Hz, 2H), 3.57 (t, J = 4.56 Hz, 4H), 2.41-2.47 ( m, 2H), 2.37 (br s, 4H), 1.97 (dd,

J = 13.94, 7.35 Hz, 2H); LCMS (LCMS Method D): retention time = 0.51 min, [M + H] + = 344.1.

Stage 2:

(E) -4,4 '- (but-2-ene-1,4-diylbis (azandiyl)) bis (3- (3-morpholinopropoxy) -5-nitrobenzamide)

To a suspension of (E) -but-2-en-1,4-diamine dihydrochloride (171 mg, 1.07 mmol) and 4-chloro-3- (3morpholinopropoxy) -5-nitrobenzamide (630 mg, 1.65 mmol) in EtOH (4 ml) DIEA (1.0 ml, 5.8 mmol) was added. The reaction mixture was heated at 120 ° C. in a heating block and additional (E) -but-2-ene-1,4-diamine dihydrochloride (30 mg, 0.19 mmol) was added after 47 hours. Heating was continued at 120 ° C for about 3 days, then the reaction mixture was dry loaded and purified on silica gel (ISCO-Rf 120 g column) eluting with 0-40% MeOH in DCM to give the title compound (130 mg, 0.186 mmol , 11% yield) as a light orange solid. ¹ H NMR (400

MHz, DMSO-46) δ ppm 8.18 (d, J = 1.77 Hz, 2H), 8.04 (br s, 2H), 7.76-7.85 (m, 2H) J = 1 , 52 Hz, 2H), 7.35 (br s, 2H), 5.63 (br s, 2H), 4.13 (br s, 4H), 4.01 (t, J = 6, 34 Hz, 4H), 3.55 (t, J = 4.56 Hz, 8H), 2.27-2.42 (m, 12H), 1.86 (t, J = 6.72 Hz, 4H) ; LCMS (LCMS Method D): retention time = 0.53 min, [M + H] + = 701.4623.

Stage 3:

(E) -4,4 '- (but-2-en-1,4-diylbis (azandiyl)) bis (3-amino-5- (3-morpholinopropoxy) benzamide) dihydrochloride

To tin (II) chloride (40.6 mg, 0.244 mmol) in a concentrated aqueous solution of HCl (8 92 μL, 10.7 mmol) was added (E) -4,4 '- (but-2-en-1,4 -diylbis (azandiyl)) bis (3- (3-morpholinopropoxy) -5nitrobenzamide) (15 mg, 0.021 mmol). After 20 minutes, the reaction mixture was allowed to cool in the refrigerator, and after 15 minutes, the resulting solid was collected by filtration to give the title compound (12 mg, 0.017 mmol, 79% yield) as a white solid. 1H NMR (400 MHz, DMSOX6) δ ppm 11.07 (brs, 2H), 7.99 (brs, 2H), 7.41 (brs, 2H), 7.05 (s, 2 H), 6.91 (br s, 2H), 5.94 (br s, 2H), 3.97-4.08 (m, 8H), 3.78-3.88 (m, 8H) , 3.48 (d, J = 12.17 Hz, 4H), 3.35-3.34 (m, 4H), 3.053.19 (m, 4H), 2.21 (bs, 4H); LCMS (LCMS Method D): retention time = 0.34 min, [M / 2 + H] + = 321.3990.

Stage 4:

(E) -1, G- (but-2-en-1,4-diyl) bis (2-amino-7- (3-morpholinopropoxy) -1H-benzoSimidazole-5carboxamide) dihydrobromide

- 74 036435

To a solution of (E) -4,4 '- (but-2-ene-1,4-diylbis (azandiyl)) bis (3-amino-5- (3-morpholinopropoxy) benzamide) dihydrochloride (102 mg, 0.143 mmol) in water (1.4 ml) was added cyanogen bromide (136 mg, 1.29 mmol). After 2 days at room temperature, the reaction mixture was added dropwise to acetonitrile (about 100 mL) and the resulting white solid was collected by filtration to give the title compound (76 mg, 0.0 9 mmol, 62% yield). ¹ H NMR (400 MHz, DMSO-de) δ ppm 13.21 (brs, 2H), 11.16 (brs, 2H), 8.88 (brs, 2H), 8.13 ( br s, 2H), 7.55 (s, 2H), 7.50 (br s, 2H), 7.42 (s, 2H), 5.71 (br s, 2H), 4.91 (brs, 4H), 4.13 (brs, 4H), 3.97 (brs, 4H), 3.82 (brs, 4H), 3.293.40 (m, 4H), 3.17 (br s, 4H), 2.99-3.09 (m, 4H) 2.08 (br s, 4H); LCMS (LCMS Method D): retention time = 0.2-8 min, [M + H] + = 691, 6058.

Stage 5:

(E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy ) -1H-benzo ^] imidazole-5-carboxamide)

To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (133 mg, 0.862 mmol), HATU (328 mg, 0.862 mmol) and HOBt (66.0 mg, 0.431 mmol) in Ν, Ν- dimethylformamide (DMF) (1150 μL) was added a suspension of (E) -1, G- (but-2-en-1,4-diyl) bis (2-amino-7- (3-morpholinopropoxy) -1H-benzo ^] imidazole-5carboxamide) dihydrobromide (209 mg, 0.245 mmol) and TEA (240 μl, 1.724 mmol) in Ν,-dimethylformamide (DMF) (4598 μl). The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness under a stream of nitrogen. The resulting residue was dissolved in methanol and dry loaded onto silica gel for purification on a 40 g ISCO-Rf column, 0-50% methanol, DCM. The target fractions were concentrated to dryness to give (E) -1,1'- (but-2-en-1,4-diyl) bis (2 (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) - 7- (3-morpholinopropoxy) - 1H-benzo (^] imidazole-5-carboxamide) (112 mg, 0.115 mmol, 47% yield) as an off-white solid. 1H NMR (400 MHz, DMSO-d ^) δ ppm 12.84 (br s, 2H), 7.98 (br s, 2H), 7.66 (d, J = 1.01 Hz, 2H) , 7.36, s, 2H), 7.25 (s, 2H), 6.58 (s, 2H), 5.82 (br s, 2H), 4.92 (br s, 4H), 4.57 (q, J = 7.10 Hz, 4H), 3.85 (t, J = 5, 96 Hz, 4H), 3.45 (t, J = 4.18 Hz, 8H), 2, 09-2.24 (m, 18H), 1.54 (t, J = 6.72 Hz, 4H), 1.32 (t, J = 7.10 Hz, 6H); LCMS (LCMS Method D): retention time = 0.65 min, [M + H] + = 963.938.

Example 13 (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) but -2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) -1Нbenzo ^] imidazole-5-carboxamide

- 75 036435

Stage 1:

K (E) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazol-1 yl) but-2 -in-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1 H-benzo ^] imidazole-5-carboxamide (17 mg, 0.023 mmol) in THF (3 ml) was added triethylamine (9.5 μl, 0.068 mmol). After 10 min at room temperature, methanesulfonyl chloride (2.1 μl, 0.027 mmol) was added. After 2 h, LCMS results showed the presence of (E) -3- ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H -benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-7-yl) hydroxy) propylmethanesulfonate and the reaction mixture was used directly in the next reaction. LCMS (LCMS Method D): retention time = 0.80 min, [M + H] + = 751, 6010.

Stage 2:

To a solution of (E) -3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1 H-benzo ^] imidazole -1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1-pyrazole-5-carboxamido) -1 H-benzo (^] imidazol-7-yl) oxy) propylmethanesulfonate (18 mg, 0.022 mmol) in THF (5 ml) was added morpholine (9.5 μl, 0.11 mmol) and K2CO3 (9.0 mg, 0.0 65 mmol). After 5 h at room temperature, the reaction mixture was heated to 45 ° C for 2 h and then concentrated. The residue was purified on silica gel eluting with 0-20% MeOH in DCM to give the title compound (7 mg, 9 μmol, 39% yield). ¹ H NMR (400 MHz, MeOH-d4) δ ppm 7.99 (d, J = 1.27 Hz, 1H), 7.73 (dd, J = 8.36, 1.52 Hz, 1H), 7 , 59 (d, J = 1.27 Hz, 1 H), 7.36 (d, J = 8.62 Hz, 1H), 7.28 (d, J = 1.27 Hz, 1H), 6, 64 (s, 1H), 6.57 (s, 1H), 5.92-6.05, 1H), 5.73-5.88 (m, 1H), 4.51-4.71 (m , 4H), 4.00 (t, J = 6.21 Hz, 2H), 3.56-3.67 (m, 8H) 2.46 (m, 6H), 2.22 (d, J = 10 , 39 Hz, 6H), 1.83 (dt, J = 14.19, 6.84 Hz, 2H), 1.26-1.44 (m, 6H); LCMS (LCMS Method D): retention time = 0.7-3 min, [M / 2 + H] ⁺ = 410, 9876.

The compound obtained by the above method may be in tautomeric or isomeric form, for example in the form (E) -1 - ((E) -4 - ((E) -5-carbamoyl-2 - ((1-ethyl-3-methyl -1Н-pyrazole-5carbonyl) imino) -2,3-dihydro-1 N-benzo (^] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl -1 H-pyrazole5-carbonyl) imine) -7- (3-morpholinopropoxy) -2,3-dihydro-1H-benzo (^] imidazole-5-carboxamide

or ^) - 1 - ((E) -4 - (^) - 5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -2,3-dihydro- 76 036435

N-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1 N-pyrazole-5-carbonyl) imino) -7- ( 3 morpholinopropoxy) -2,3-dihydro-1H-benzo ^] imidazole-5-carboxamide

Example 14. (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) -1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 7-methoxy-1H-benzo ^] imidazole-5-carboxamide tris hydrochloride

Stage 1:

(E) -4 - ((4 - ((4-carbamoyl-2- (3-morpholinopropoxy) -6-nitrophenyl) amino) but-2-en-1-yl) amino) -3methoxy-5-nitrobenzamide

- 77 036435

(E) -4- ((4-Aminobut-2-en-1-yl) amino) -3-methoxy-5-nitrobenzamide hydrochloride (1.7 g, 5.37 mmol), 4-chloro-3- ( 3-morpholinopropoxy) -5-nitrobenzamide (1.655 g, 4.81 mmol), i-PrOH (15 ml) and DIPEA (2.94 ml, 16.85 mmol) were divided into two 24 ml vials, then the vials were closed and heated to 120 ° C for 42 h. The solid was isolated by filtration, washed with i-PrOH (2x3 ml) to give (E) -4 - ((4 - ((4-carbamoyl-2- (3-morpholinopropoxy) -6 -nitrophenyl) amino) but-2-en-1-yl) amino) -3methoxy-5-nitrobenzamide (1.95 g, 2.79 mmol, 51.9% yield) as a brick red solid. LCMS (LCMS Method K): retention time = 0.60 min, [M + H] + = 588.2.

Stage 2:

(E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6- (3-morpholinopropoxy) phenyl) amino) but-2-en-1yl) amino) -5-methoxybenzamide

K (E) -4 - ((4-carbamoyl-2- (3-morpholinopropoxy) -6-nitrophenyl) amino) but-2-en-1-yl) amino) -3methoxy-5-nitrobenzamide (4.6 g , 6.65 mmol) in MeOH (83.0 ml), sodium hydrosulfurous acid (19.08 g, 93.0 mmol) in water (70 ml) was added at room temperature. Solid sodium bicarbonate (24 g) was added after 15 minutes. After 10 min, the reaction mixture was filtered and the solid was washed with MeOH (4 x 20 ml). The combined filtrates were concentrated on celite and purified by dry loading onto silica gel (80 g Gold column) eluting with 2-40% (10: 1 MeOH: NH4OH aqueous solution) in DCM to give the title compound (1.81 g, 3.26 mmol, yield 49%) in the form of a dark yellow film. 1H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 7.64 (br s, 2H), 6.99 (br s, 2H), 6.85 (dd, J = 5.07, 1.77 Hz , 2H) (dd, J = 4.31, 1.77 Hz, 2H), 5.63-5.72 (m, 2H), 4.66 (d, J = 8.11 Hz, 4H), 3 , 96 (t, J = 6.21 Hz, 2H) 3.74 (s, 3H), 3.51-3.60 (m, 6H), 3.17 (br s, 4H), 2.43 (t, J = 7.10 Hz, 2H), 2.35 (br s, 4H), 1.87 (t, J = 6.72 Hz, 2H); LCMS (LCMS Method K): retention time = 0.37 min, [M + H] ⁺ = 528.4.

Stage 3:

(E) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) 1 H-benzo ^] imidazole- 1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1Hbenzo ^] imidazole-5-carboxamide tris- hydrochloride

- 78 036435

K (E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6- (3-morpholinopropoxy) phenyl) amino) -but-2-en-1yl) amino) -5- methoxybenzamide (368 mg, 0.697 mmol) in DMF (6.97 ml) at 0 ° C was added a 0.4 M solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate in dioxane (2.0 ml, 0 , 80 mmol). After about 10 minutes, another portion of a 0.4 M solution of 1-ethyl-3-methyl-1H-pyrazole-5carbonyl isothiocyanate in dioxane (0.5 ml, 0.20 mmol) was added, followed by the addition of the final portion (0, 5 ml, 0.20 mmol). After 35 min total reaction time, EDC (334 mg, 1.74 mmol) was added followed by triethylamine (0.486 mL, 3.49 mmol). The mixture was allowed to warm to room temperature and stirred overnight (about 14 hours). The reaction was quenched with a 3: 1 mixture of water: saturated aqueous NH4Cl (40 ml) and the reaction mixture was extracted with a mixture of chloroform: ethanol (2 x 40 ml) 3: 1. The combined organic phases were washed with water (20 ml), dried over MgSO ₄ and concentrated. The resulting residue was purified on silica gel (40 g Gold column) eluting with 2-40% (10: 1 MeOH: NH4OH aqueous solution) in DCM to give the pure material as the free base. This product was dissolved in MeOH and treated with 4M HCl in dioxane (0.35 ml, 1.40 mmol), then concentrated. The residue was dissolved in MeCN-water and lyophilized to afford the title compound (403.6 mg, 0.421 mmol, 60% yield) as an off-white solid. ¹ H NMR (400 MHz, methano.i-d4) δ ppm 7.70 (dd, J = 2.66, 1.14 Hz, 2H), 7.42 (d, J = 1.27 Hz, 2H) , 6.72 (d, J = 3.04 Hz, 2H), 5.79-6.12 (m, 2H), 5.19 (dd, J = 11.03, 5.45 Hz, 4H), 4.61-4.81 (m, 4H), 4.00-4.25 (m, 4H), 3.79-3.96 (m, 5H), 3.45 (d, J = 12.42 Hz, 2H), 3.28-3.36 (m, 2H), 3.14 (td, J = 12.23, 3.68 Hz, 2H), 2.28 (s, 6 H), 2, 07-2.25 (m, 2H), 1.46 (td, J = 7.10, 3.80 Hz, 6H); LCMS (LCMS Method K): retention time = 0.68 min, [M + H] + = 850.6.

The compound obtained by the above method may be in tautomeric or isomeric form, for example, in the form (E) -1 - ((E) -4 - (^) - 5-carbamoyl-2 - ((1-ethyl-3-methyl- 1Н-pyrazole-5-carbonyl) imino) -7 - (3-morpholinopropoxy) -2,3-dihydro-1Н-benzo (d) imidazol-1-yl) but-2-en-1-yl) -2- ( (1-ethyl3-methyl-1 H-pyrazole-5-carbonyl) imino) -7-methoxy-2,3-dihydro-1 H-benzo ^] imidazole-5-tris-hydrochloride carboxamide or (Z) -1 - ((E) -4 - ((Z) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5 -carbonyl) imino) -7- (3-morpholinopropoxy) -2.3 -dihydro- 1H-benzo ^] imidazol-1-yl) but-2-en-1 -yl) -2 - ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl) imino) -7-methoxy- 2,3-dihydro-1H-benzo ^] imidazole-5-tris-hydrochloride carboxamide

Example 15 (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7 - ((4-methoxybenzyl) oxy) -1HbenzoI imidazole-5-carboxamide

- 79 036435

PMBCI CsnCO

Stage 1:

4-chloro-3 - ((4-methoxybenzyl) oxy) -5-nitrobenzamide

N32S2O4 NH.OH

- 80 036435

4-Chloro-3-hydroxy-5-nitrobenzamide (942 mg, 4.35 mmol) was dissolved in DMF (7 ml), Cs ₂ CO ₃ (1.559 g, 4.78 mmol) was added followed by the addition of 4-methoxybenzyl chloride (0.622 ml, 4.57 mmol) and the reaction mixture was stirred for 24 h at room temperature. With vigorous stirring, water (15 ml) was added dropwise and the resulting solid was stirred for 5 min, collected by filtration and washed with water to give the title compound (1.26 g, 3.74 mmol, 82% yield) as a light - orange solid. ¹ H NMR (400 MHz,

CDCl3) δ ppm 7.80 (d, J = 1.8 Hz, 1H), 7.76 (d, J = 1.8 Hz, 1H), 7.43 (d, J = 8.6 Hz, 2H ) d, J = 8.6 Hz, 2H), 6.13 (br s, 1H), 5.82 (br s, 1H), 5.25 (s, 2H), 3.87 (s, 3H); LCMS (LCMS Method D): retention time = 1.03, [M + H] + = 337, 1.

Stage 2:

(E) -1- (4 - ((4-carbamoyl-2- ((4-methoxybenzyl) oxy) -6-nitrophenyl) amino) but-2-en-1-yl) -2- (1-ethyl- 3methyl-1 H-pyrazole-5-carboxamido) -1 H-benzoI imidazole-5-carboxamide

To the mixture (E) -1 - (4-aminobut-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazole -5-carboxamide hydrochloride (1.543 g, 3.69 mmol, intermediate 6) and TEA (1.871 ml, 13.42 mmol), stirred in EtOH (7 ml) for 5 min, 4-chloro-3- ( (4methoxybenzyl) oxy) -5-nitrobenzamide (1.13 g, 3.36 mmol) and the mixture was heated in a sealed microwave vial at 120 ° C for 18 h. After cooling to room temperature, the mixture was diluted with DCM (50 ml ) and water (50 ml) and a dark residue appeared. The layers were separated and the residue was combined with the organic phases and concentrated. To the crude mixture was added 10% MeOH in DCM and the resulting solid was collected by filtration and washed with DCM. To the concentrated filtrate was again added 10% MeOH in DCM and the resulting solid was collected by filtration and washed with DCM. Both batches of solid were combined to give the title compound (559 mg, 0.82 mmol, 22% yield) as an orange solid. ¹ H NMR (400 MHz, DMSO-f) δ ppm 12.86 (br s, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.93-8.05 (m, 3H), 7.86 (t, J (D, J = 8.4, 1.3 Hz, 1H), 7.62 (s, 1H) (m, 1H), 4.93 (s, 2H), 4.75 Hz, 2H), 4.58 (q, J = 7.1 Hz, 2H), 4.05 (t, J = 5.6 Hz, 2H), 3.69 (s, 3H), 2 , 16 (s, 3H), 1.31 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.98, [M + H] + = 682.5.

Stage 3:

(E) -1- (4 - ((2-amino-4-carbamoyl-6 - ((4-methoxybenzyl) oxy) phenyl) amino) but-2-en-1-yl) -2- (1-ethyl -3methyl-1 H-pyrazole-5-carboxamido) -1 H-benzoI imidazole-5-carboxamide

K (E) -1- (4- ((4-carbamoyl-2- ((4-methoxybenzyl) oxy) -6-nitrophenyl) amino) but-2-en-1-yl) -2- (1ethyl-3 -methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide (557 mg, 0.817 mmol) in DMF (10 ml) was added ammonium hydroxide (1.136 ml, 8.17 mmol) followed by the addition of dropwise sodium hydrogen sulfite (837 mg, 4.09 mmol) in water (5 ml). After 1 h at room temperature, the reaction mixture was diluted with EtOAc and water. The aqueous extract was extracted with EtOAc, the combined organic fractions were washed with saturated NH ₄ Cl solution and brine and concentrated to give the title compound (335 mg, 0.51 mmol, 57% yield) as

- 81 036435 orange solid. 1H NMR (400 MHz, DMSO-dg) δ ppm 12.85 (br s, 1H), 8.00 (s, 2H), 7.75 (dd, J = 8.4, 1.5 Hz, 1H ), 7.66 1H), 7.36 (br s, 2H), 7.25-7.31 (m, 4H), 7.03 (br s, 1H), 6.83-6.91 (m, 4H),

6.64 (s, 1H) (M, 1H), 4.89 (s, 2H), 4.78 (d, J = 5, l Hz, 2H), 4.69 (br s, J = 7 , 0 Hz, 2H), 4.59 (q, J = 7.0 Hz, 2H),

3.91 (t, J = 7.0 Hz, 1H), 3.71 (s, 3H), 3.56 (br m, 2H), 2.17 (s, 3H), 1.32 (t , J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.76, [M + H] + = 652.5.

Stage 4:

(E) -2-amino-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) but -2-en-1-yl) -7 - ((4-methoxybenzyl) oxy) -1Н-benzo ^] imidazole-5-carboxamide dihydrobromide

To the suspension (E) -1 - (4 - ((2-amino-4-carbamoyl-6 - ((4-methoxybenzyl) oxy) phenyl) amino) but-2-en-1 yl) -2- (1 - ethyl 3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazole-5-carboxamide (333 mg, 0.460 mmol) in MeOH (3 ml), cyanogen bromide (97 mg, 0.920 mmol) was added and the reaction mixture was stirred at room temperature for 2 hours. The resulting solid was collected by filtration and washed with MeOH to give the title compound (235 mg, 0.28 mmol, 58% yield) as a light orange solid. 1H NMR (400 MHz, DMSO-dg) δ ppm 13.07 (brs, 1H), 12.90 (s, 1H), 8.70 (brs, 1H), 8.09 (brs , 7.98-8.04 (m, 2H), 7.73 (d, J = 8.6 Hz, 1H), 7.51 (s, 2H), 7.46 (brs, 1H), 7.32-7.40 (m, 2H) d, J = 8.6 Hz, 2H), 6.77 (d, J = 8.4 Hz, 2H), 6.54 (s, 1H), 5 , 83-5.92 (m, 1H), 5.53-5.62 (m, 1H), 2H), 4.79 (s, 2H), 4.78 (s, 2H), 4.53 ( q, J = 7.2 Hz, 2H), 3.66 (s, 3H) 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.72, [M + H] + = 677.5.

Stage 5:

(E) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazol-1 yl) but-2- en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7 - ((4-methoxybenzyl) oxy) -1Hbenzo | c1 | imidazole-5-carboxamide

A solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (51.4 mg, 0.333 mmol) and CDI (63.1 mg, 0.389 mmol) in DMF (3 ml) was stirred at 60 ° C for 10 min, then (E) -1- (4- (2amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido) -7 - ((4-methoxybenzyl) oxy) -1H-benzo ^] imidazole-5-carboxamide dihydrobromide (233 mg, 0.278 mmol) and DIPEA (0.194 ml, 1.111 mmol ). After heating at 90 ° C for 4 hours, additional CDI (20 mg) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (20 mg) were added and heating was continued for 1.5 hours. cooling to room temperature and adding small pieces of ice with stirring was added dropwise water (5 ml). The resulting solid was collected by filtration and washed with water to give the title compound (225 mg, 0.27 mmol, 95% yield) as a light brown solid. 1H NMR (400 MHz, DMSO-dg) δ ppm 12.88 (br s, 2H), 8.00 (m, 2H), 7.64-7.72 (m, 2H), 7.46 (br s, 1H), 7.20 (d, J = 7.9 Hz, 2H), 6.75 (d, J = 7.6 Hz, 2H), 6.54 (s, 2H), 5.94 (m, 1H), 5.49, 1H), 4.97 (s, 2H), 4.85 (br s, 2H), 4.78 (br s, 2H), 4.53 (br. s, 4H), 3.65 (s, 3H) s, 3H), 2.10 (s, 3H), 1.22-1.33 (m, 6H); LCMS (LC-MS Method F) RT 82 036435 live = 2.27 min, [M + H] + = 813.9.

Example 16 (E) -1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -1H-benzo [d] imidazol-1-yl) but -2-yl-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-hydroxy-1H-benzoSimidazole-5-carboxamide

K (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoH | imidazol-1yl) 2-en-1-yl ) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7 - ((4-methoxybenzyl) oxy) -1H-benzoHimidazole-5-carboxamide (210 mg, 0.258 mmol) in dioxane ( 2 ml) was added 4N HCl solution in dioxane (0.258 ml, 1.03 mmol). After 30 min at room temperature, the reaction mixture was heated to 50 ° C for 18 h. An additional 4N HCl solution in dioxane (0.2 ml) was added and after heating for an additional 5 h, the reaction mixture was cooled and the resulting solid collected by filtration and washed with DCM to give the title compound (168 mg, 0.229 mmol, 81% yield) as a light brown solid. 'H NMR (400 MHz, MeOH-d ₄ ) δ ppm 8.01 (d, J = 1.3 Hz, 1H), 7.82 (dd, J = 8.5, 1.6 Hz, 1H), 7.49-7.55 (m, 2H) 7.29 (d, J = 1.5 Hz, 1H), 6.68 (s, 1H), 6.58 (s, 1H), 6.08- 6.18 (m, 1H), 5.89-5.99 (m, 1H), 5.30 (d, J = 5.6Hz, 2H), 4.98-5.04 (m, 2H) , 4.65 (dq, J = 14.6, 7.2 Hz, 4H), 2.23 (s, 3H), 2.20 (s, 3H), 1.37-1.46 (m, 6H); LCMS (LCMS Method F): retention time = 1.7-3 min, [M + H] + = 693.4.

Example 17 1,1 '- (2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazole-5-carboxamide)

- 83 036435

Stage 1:

4,4 '- ((2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (azandiyl)) bis (3-nitrobenzamide)

To 2,2,3,3-tetrafluorobutane-1,4-diamine (intermediate 4) (1.25 g, 7.81 mmol) and potassium carbonate (3.24 g, 23.4 mmol) in DMF (50 ml) 4-fluoro-3-nitrobenzamide (3.59 g, 19.5 mmol) was added at room temperature over 5 min and the reaction mixture was stirred overnight. The mixture was quenched with water and the resulting solid was collected by filtration and triturated with MeOH to give the title compound (600 mg, 1.23 mmol, 16% yield) as a yellow solid. LCMS (LCMS Method A): retention time = 1.3 67 min, [M + H] + = 489.0.

Stage 2:

4,4'- ((2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (azandiyl)) bis (3-aminobenzamide)

- 84 036435

4,4 '- ((2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (azavdiyl)) bis (3-nitrobenzamvd) (1.15 g, 2.36 mmol) and Pd on carbon ( 0.251 g, 2.36 mmol) in MeOH (100 ml) was stirred under a hydrogen atmosphere at 30 ° C overnight. The reaction mixture was filtered and the filtrate was concentrated to give the title compound (250 mg, 0.584 mmol, 25% yield). LCMS (LCMS Method A): retention time = 1.165 min, [M + H] + = 429.1.

Stage 3:

1, D- (2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (2-amino-1H-benzoU] imidazole-5-carboxamide)

To 4,4 '- ((2,2,3,3-tetrafluorobutane-1,4-diyl) bis (azandiyl)) bis (3-aminobenzamide) (20 mg, 0.047 mmol) in MeOH (1 ml) and water (2 ml) cyanogen bromide (29.7 mg, 0.280 mmol) was added and the reaction mixture was stirred at 30 ° C overnight. The MeOH was removed in vacuo and the resulting solid was collected by filtration to give the title compound (15 mg, 0.031 mmol, 67% yield). LCMS (LCMS Method A): retention time = 0.629 min, [M + H] + = 479.0.

Stage 4:

1,1 '- (2,2,3,3-Tetrafluorobutane-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1НbenzoShimidazole-5-carboxamide)

To HATU (763 mg, 2.01 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (227 mg, 1.47 mmol) in DMF (20 ml) at room temperature was added EDC (385 mg, 2.01 mmol), 1,1 '- (2,2,3,3 tetrafluorobutane-1,4-diyl) bis (2-amino-1H-benzoU] imidazole-5-carboxamide) (320 mg, 0.667 mmol ) and DIEA (0.467 ml, 2.68 mmol) in one serving. The reaction mixture was heated to 70 ° C for 12 h, concentrated and purified to give the title compound (8 mg, 0.01 mmol, 2% yield). ¹ H NMR (400 MHz, DMSO-06) δ ppm 13.05 (s, 2H), 8.01 (d, J = 8.6 Hz, 4H), 7.81 (d, J = 8.2 Hz , 2H), 7.53 (d, J = 8.3 Hz, 2H), 7.38 (s, 2H), 6.73 (s, 2H), 5.32 (t, J = 16.0 Hz , 4H), 4.59 (dd, J = 14.0, 6.9 Hz, 4 H), 2.06 (s, 6H), 1.33 (t, J = 7.1 Hz, 6H); LCMS (LCMS Method A): retention time = 1.367 min, [M + H] ⁺ = 751.1.

Example 18 Di-tert-butyl (3 - (((2) -6-carbamoyl-3 - ((E) -4 - ((2) -5-carbamoyl-2 - ((1-ethyl-3-methyl -1H pyrazole-5-carbonyl) imino) -7-methoxy-2,3-dihydro-1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl- 3- 85 036435 methyl-1 H-pyrazole-5-carbonyl) imino) -2,3-dihydro-1 H-6eH3o [d] imidazol-4-yl) oxy) propyl) phosphate

Di-tert-butyl (3 - (((2) -6-carbamoyl-3 - ((E) -4 - ((2) -5-carbamoyl-2 - ((1-ethyl-3 -methyl-1 H -pyrazole-5carbonyl) imino) -7 -methoxy-2,3-dihydro-1 H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3 -methyl1H-pyrazole-5-carbonyl) imino) -2,3-dihydro-III-benzo (^] imidazol-4-yl) oxy) propyl) phosphate

Suspension (E) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1 H-benzoI imidazole- 1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7 methoxy-1H-benzo ^] imidazole-5-carboxamide ( 1.00 g, 1.28 mmol) and a 0.45 M solution of 2H-tetrazole in acetonitrile (14.2 ml, 6.40 mmol) in DMF (5 ml) were concentrated on a rotary evaporator to remove acetonitrile. The resulting heterogeneous mixture in DMF was cooled to 0 ° C, then a solution of tert-butyldiisopropylphosphoramidite (1.617 ml, 5.12 mmol) in 5 ml of DMF was added. Shortly after the addition, the solution became homogeneous but became heterogeneous again when the reaction mixture was stirred at room temperature for an additional 2 h. The temperature was lowered to 0 ° C and H ₂ O ₂ (30 wt% in water, 2.62 ml, 25.6 mmol). After stirring for 20 min, another 10 eq of H ₂ O _{2 was} added and the reaction mixture was stirred until homogeneous (30 min). To 200 ml of water was added

- 86 036435 a portion of 2 ml of an aqueous solution of NaHCO ₃ and Na ₂ S ₂ O ₃ (0.4 M NaHCO ₃ solution, 2M Na ₂ S ₂ O ₃ solution). When the reaction mixture was poured into this solution, a precipitate formed. The precipitate was then collected on a filter, dissolved in 200 ml THF, dried over MgSO ₄ and concentrated to give the title compound as an off-white solid (1.1 g, 1.13 mmol, 88% yield). 1H NMR (400 MHz, DMSO-A) δ ppm 12.8 (s, 2H), 10.2 (s, 1H), 7.98 (m, 2H), 7.65 (d, J = 2.5 Hz, 2H), 7.34 (m, 4H), 6.51 (d, J = 2.5 Hz, 2H), 5.83 (m, 2H), 4.91 (m, 4H), 4, 52 (m, 4H), 4.09 (m, 2H), 3.93 (m, 2H), 3.74 (s, 3H), 3.60 (m, 2H), 2.11 (s, 6H ), 1.90 (m, 2H), 1.76 (m, 2H), 1.4-1.3 (m, 18H, 1.27 (m, 6H), LC-MS (LC-MS method I ): retention time = 1.0 9 min, [M + H] + = 973.3.

Example 19 3 - (((2) -6-Carbamoyl-3 - ((E) -4 - ((2) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl ) imino) -7-methoxy-2,3-dihydro-1H-benzoY] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl1H-pyrazole-5 -carbonyl) imino) -2,3-dihydro-1H-benzoIimidazol-4-yl) oxy) propyldihydrogen phosphate

To di-tert-butyl (3 - (((Z) -6-carbamoyl-3 - ((E) -4 - ((Z) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1Н -pyrαzol-5carbonyl) imino) -7-methoxy-2,3-dihydro-1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl1H -pyrazole-5-carbonyl) imino) -2,3-dihydro-1H-benzo (^] imidazol-4-yl) oxy) propyl) phosphate (18 mg, 0.018 mmol) suspended in dioxane (1 ml), at room temperature was added 4N solution of HCl in dioxane (0.02-8 ml, 0.11 mmol). Some precipitate formed immediately. The reaction mixture was stirred for 2 h and additional 4N HCl in dioxane (0.028 mL, 0.11 mmol) was added. After 2 hours, the reaction mixture was placed in a freezer and after 16 hours, the reaction mixture was diluted with diethyl ether. The pH of the mixture was adjusted to 2-3 with concentrated ammonium hydroxide. The precipitate was collected by filtration and washed with diethyl ether to give the title compound (15 mg, 0.017 mmol, 92% yield) as a white solid. ¹ H NMR (600 MHz, DMSO-de) δ ppm 12.85 (br s, 1H), 8.02 (br d, J = 6.6 Hz, 2H), 7.65 (d, J = 5.7 Hz, 2H) 7.41 (m, 2H), 7.34 (br d, J = 10.6 Hz, 2H), 6.51 (d, J = 12.8 Hz, 2H), 5.74-5.89 (m, 2H), 4.92 (br.dd, J = 12.0, 4.9 Hz, 4H), 4.50 (quint, J = 7.0 Hz, 4H) , 4.10 (br t, J = 6.1 Hz, 2H), 3.91-3.94 (m, 2H), 3.75 (s, 3H), 2.10 (d, J = 3 , l Hz, 6H), 1.84-1.93 (m, 2H), 1.22-1.28 (m, 6H); LCMS (LCMS Method I): retention time = 0.68 min, [M + H] + = 861.2.

The compound obtained by the above method may be in tautomeric or isomeric form, for example, in the form (E) -3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl- 1H-pyrazole-5carboxamido) -7-methoxy-1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) - 1H-benzoI imidazol-7-yl) oxy) propyldihydrogen phosphate

or 3 - (((E) -6-carbamoyl-3 - ((E) -4 - ((E) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl ) imino) -7-methoxy-2,3-dihydro-1 H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl- 1H-pyrazole-5carbonyl) imino) -2,3-dihydro-1H-benzoU] imidazol-4-yl) oxy) propyldihydrogen phosphate

- 87 036435

Example 20.

Stage 8:

(E) -7-bromo-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-methoxypropoxy) -1H-benzo ^ ] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [c1] imidazole-5-carboxamide

- 88 036435

Stage 1:

4-Chloro-3 - (3-methoxypropoxy) -5-nitrobenzamide

To a suspension of 4-chloro-3-hydroxy-5-nitrobenzamide (1.00 g, 4.62 mmol) in DMF (15 ml) was added 1-bromo-3-methoxypropane (1.06 g, 6.93 mmol) and K ₂ CO ₃ (1.91 mg, 13.9 mmol). The reaction mixture was stirred at 60 ° C in a sealed tube. After 3 hours, the reaction mixture was cooled to room temperature and poured into water. The resulting light yellow precipitate was collected by filtration and washed with diethyl ether to give the title compound (1.1 g, 3.8 mmol, 83% yield). LCMS (LCMS Method D): retention time = 0.84 min, [M + H] + = 289.0.

Stage 2:

(E) -tert-butyl (4 - ((4-carbamoyl-2- (3-methoxypropoxy) -6-nitrophenyl) amino) but-2-en-1yl) carbamate

- 89 036435

To a suspension of 4-chloro-3-methoxy-5-nitrobenzamide (1.50 g, 6.50 mmol) in EtOH (25 ml) was added (E) tert-butyl (4-aminobut-2-en-1-yl) carbamate (1.45 g, 7.81 mmol) and DIEA (3.41 ml, 19.5 mmol). The reaction mixture was heated to 120 ° C in a sealed tube overnight and left to cool to room temperature. The resulting orange precipitate was collected by filtration and washed with EtOH to give the title compound (2.1 g, 5.5 mmol, 85% yield). LCMS (LCMS Method D): retention time = 0.96 min, [M + H] + = 439.2.

Stage 3:

(E) -4 - ((4-aminobut-2-en-1-yl) amino) -3- (3-methoxypropoxy) -5-nitrobenzamide dihydrochloride

K (E) -tert-butyl (4 - ((4-carbamoyl-2- (3-methoxypropoxy) -6-nitrophenyl) amino) but-2-en-1-yl) carbamate (1.43 g, 3, 26 mmol) in dioxane (10 ml) was added 4N HCl solution in dioxane (12.2 ml, 48.9 mmol). After 1 h at room temperature, the reaction mixture was concentrated and the residue triturated with diethyl ether to give the title compound (1.3 g, 3.1 mmol, 96% yield). LCMS (LCMS Method D): retention time = 0.52 min, [M + H] + = 339.2.

Stage 4:

(E) -3-bromo-4 - ((4 - ((4-carbamoyl-2- (3-methoxypropoxy) -6-nitrophenyl) amino) but-2-en-1-yl) amino) 5-nitrobenzamide

To a solution of (E) -4- ((4-aminobut-2-en-1-yl) amino) -3- (3-methoxypropoxy) -5-nitrobenzamide dihydrochloride (361 mg, 0.878 mmol) in DMF (5 ml) 3-bromo-4-fluoro-5-nitrobenzamide (220 mg, 0.836 mmol) and TEA (0.47 ml, 3.4 mmol) were added. After stirring at room temperature overnight, water (20 ml) was added and the resulting light brown solid was collected by filtration to give the title compound (475 mg, 0.719 mmol, 86% yield). LCMS (LCMS Method D): retention time = 0.91 min, [M + H] + = 583.2.

Stage 5:

(E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6- (3-methoxypropoxy) phenyl) amino) but-2-en-1yl) amino) -5-bromobenzamide

To a solution of (E) -3-bromo-4 - ((4 - ((4-carbamoyl-2- (3-methoxypropoxy) -6-nitrophenyl) amino) but-2-ene-1- 90 036435 yl) amino) -5-nitrobenzamide (400 mg, 0.585 mmol) in concentrated aqueous 12 M HCl solution (5 ml, 60 mmol) was added tin (II) chloride (665 mg, 3.51 mmol). After 5 minutes at room temperature, a 6N aqueous NaOH solution was added to neutralize the reaction mixture, and the resulting solid was collected by filtration to give the title compound (150 mg, 0.288 mmol, 49% yield). LCMS (LCMS Method D): retention time = 0.55 min, [M + H] + = 521.2.

Stage 6:

(E) -2-amino-1 - (4- (2-amino-5-carbamoyl-7- (3-methoxypropoxy) -1H-benzo Sh | imidazol-1-yl) but-2-en1-yl) - 7-bromo-1H-benzo [d] imidazole-5-carboxamide o

about

To a solution of (E) -3-amino-4 - ((4 - ((2-amino-4-carbamoyl-6- (3-methoxypropoxy) phenyl) amino) but-2-en1-yl) amino) -5- bromobenzamide (150 mg, 0.288 mmol) in MeOH (3 ml) and DMSO (1 ml) was added cyanogen bromide (183 mg, 1.73 mmol). The reaction mixture was stirred at room temperature overnight, during which time a solid precipitated. This solid was collected by filtration to give the crude title compound (120 mg, 0.210 mmol, 73% yield). LCMS (LC-MS Method D): retention time = 0.4 7 min, [M + H] + = 573.2.

Stage 8:

(E) -7-bromo-1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-methoxypropoxy) -1H-benzo ^ imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -1Нbenzo Schimidazo l-5 -carboxamide

To 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (97 mg, 0.63 mmol) in DMF (2 ml) were added HATU (240 mg, 0.630 mmol) and HOBt (48 mg, 0.32 mmol). After 15 min at room temperature, TEA (0.18 ml, 1.3 mmol) was added followed by the addition of (E) -2-amino-1- (4- (2-amino-5-carbamoyl7- (3-methoxypropoxy) - 1H-benzo-S-imidazol-1-yl) but-2-en-1-yl) -7-bromo-1H-benzo-imidazole-5-carboxamide (120 mg, 0.210 mmol). After stirring overnight, water (5 ml) was added and the resulting light yellow precipitate was collected by filtration. This crude material was purified on silica gel (Isco column Rf 25 g, eluting with 0-2 0% MeOH in DCM) to afford the title compound (45 mg, 0.050 mmol, 24% yield). 1H NMR (400 MHz, DMSOD6) δ ppm 13.06 (brs, 1H), 12.83 (brs, 1H), 8.07 (brs, 1H), 8.00 (d, J = 8.62 Hz, 2H), 7.88-7.96 (m, 1H), 7.64 (s, 1H), 7.47 (brs, 1H), 7.28-7.39 ( m, 2H), 6.49-6.59 (m, 2H), 5.70-5.92 (m, 2H), 5.09 (bs, 2H), 4.93 (bs, 2H), 4.44-4.60 (m, 4H), 4.03 (t, J = 6.46 Hz, 2H), 3.23-3.30 (m, 2H), 3.14 (s , 3H), 2.11 (d, J = 12.17 Hz, 6H), 1.71-1.83 (m, 2H), 1.27 (q, J = 7.10 Hz, 6H); LCMS (LCMS Method D): retention time = 0.96 min, [M / 2 + H] + = 422.1.

The compound obtained by the above method may be in tautomeric or isomeric form, for example in the form (E) -7-bromo-1 - ((E) -4 - ((E) -5-carbamoyl-2 - ((1-ethyl -3-methyl-1H-pyrazole-5carbonyl) imino) -7 - (3-methoxypropoxy) -2,3-dihydro-1H-benzo (d-imidazol-1-yl) but-2-en-1-yl) -2 - ((1 ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -2,3-dihydro-1H-benzo-imidazole-5-carboxamide

- 91 036435

or (Z) -7-6poM-1- ((E) -4- (^) - 5-carbamoyl-2- ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) 7- (3methoxypropoxy) -2,3-dihydro-1 H-6eH3o [d] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1 H pyrazole 5 - carbonyl) imine) -2,3-dihydro-1 H-6eH3o [d] imidazole-5-carboxamide o

about

Example 21.

Stage 9:

Ethyl (E) -3 - (5-carbamoyl-1 - (4- (5-carbamoyl-2- (1-ethyl-3 -methyl- 1H-pyrazole-5-carboxamido) -7- (3 methoxypropoxy) -1H -benzo [d] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazole -7-yl) propanoate

A mixture of (E) -7-bromo-1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -7- (3-methoxypropoxy) -1 H -benzo ^] imidazol-1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -1Hbenzo ^] imidazole-5-carboxamide ( 50 mg, 0.059 mmol), Pd (amphos) Cl ₂ (20 mg, 0.028 mmol) and bromide (3-ethoxy-3-oxopropyl) zinc (II) (1.5 ml, 0.750 mmol) were heated in a microwave reactor at 100 ° C ... After 15 min, the reaction mixture was cooled, concentrated and the crude material was purified by reverse phase HPLC (Gilson HPLC (CH3CN / H2O 10-60%) to afford the title compound (20 mg, 0.023 mmol, 38.2% yield). 1H NMR ( 400 MHz, MeOH-d ₄ ) δ ppm 7.85 (d, J = 1.52 Hz, 1H), 7.59 (d, J = 1.01 Hz, 1H), 7.54 (d, J = 1.52 Hz, 1H) 7.32 (d, J = 1.01 Hz, 1H), 6.61 (d, J = 7.10 Hz, 2H), 5.92 (d, J = 15.72 Hz, 1H), 5.61-5.77 (m, 1H), 4.99-5.23 (m, 4H), 4.50-4.99 (m, 4H), 4.08 (t, J = 6.34 Hz, 2H), 4.00 (q, J = 7.18 Hz, 2H), 3.41 (t, J = 6.21 Hz, 2H), 3.28 (s, 3H) , 3.12 (t, J = 7.86 Hz, 2H), 2.53-2.65 (m, 2H), 2.22 (s, 6H), 1.85 (quinn, J = 6.27 Hz, 2H), 1.24-1.47 (m, 9H), 1.15 (t, J = 7.10 Hz, 3H), LC-MS (LC-MS Method K): retention time = 0, 8 7 min, [M + H] + = 865.2.

Example 22.

Stage 10:

Ethyl (E) -3 - (5-carbamoyl-1 - (4- (5-carbamoyl-2- (1-ethyl-3 -methyl- 1H-pyrazole-5-carboxamido) -7- (3 methoxypropoxy) -1H -benzo [d] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazole -7-yl) propanoic acid

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K (E) -ethyl 3 - (5-carbamoyl-1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3 methoxypropoxy) - 1H-benzoO | imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoO | imidazol-7-yl ) propanoate (15 mg, 0.014 mmol, example 21) in THF (0.3 ml) was added 1 M equiv of LiOH (0.042 ml, 0.042 mmol) in water (0.3 ml). After 1 h at room temperature, THF was removed in vacuo and a 5N aqueous HCl solution was added to neutralize the reaction mixture. The resulting white solid was collected by filtration and this crude material was purified by HPLC (Gilson column, Sunfire C18 OBD 30 x 100 mm with a gradient of 10-60% acetonitrile (0.1% TFA) / water (0.1% TFA) and a flow rate of 30 ml / min, gradient time: 15 min) to give the title compound (2.4 mg, 3 μmol, 20% yield). 'H NMR (400 MHz, methanol D ₄ ) δ ppm 7.86 (d, J = 1.52 Hz, 1H), 7.58 (dd, J = 3.55, 1.27 Hz, 2H), 7, 31 (d, J = 1.01 Hz, 1 H), 6, 60 (d, J = 5.83 Hz, 2H), 5.92 (d, J = 15.46 Hz, 1H), 5.69 (d, J = 15.46 Hz, 1H), 5.12 (br H), 5.06 (d, J = 4.82 Hz, 2H), 4.60 (dc, J = 14.61, 7.21 Hz, 4H), 4.09 (t, J = 6.46 Hz, 2H), 3.42 (t, J = 6.08 Hz, 2H), 3.28 (s, 3H), 3 , 09-3.18 (m, 2H), 2.56-2.65 (m, 2H), 2.21 (d, J = 4.82 Hz, 6H) t, J = 6.21 Hz, 2H ), 1.35 (dt, J = 12.42, 7.10 Hz, 6H); LCMS (LCMS Method D): retention time = 1.90 min, [M + H] + = 838.0.

Example 23. Methyl 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo-imidazol-1-yl) butyl) -2- (1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoSHimidazole-5carboxylate bis-trifluoroacetate

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Stage 1:

4 - ((4 - ((tert-butoxycarbonyl) amino) butyl) amino) -3-nitrobenzoic acid hydrochloride

To tert-butyl (4-aminobutyl) carbamate (4.00 g, 21.3 mmol) and potassium carbonate (8.81 g, 63.7 mmol) in DMSO (70.8 ml) at room temperature was added 4-fluoro -3-nitrobenzoic acid (3.93 g, 21.3 mmol). The reaction mixture was heated to 80 ° C for 18 h, cooled to room temperature and diluted with EtOAc and water. The mixture was vigorously stirred and carefully adjusted to pH 5 with HCl. The organic layer was separated, washed with water and brine, dried over sodium sulfate, filtered and concentrated to give the crude title compound (11.86 g, 21.3 mmol, quantitative yield) as a yellow amorphous solid, which was used without subsequent cleaning. ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.89 (br s, 1H), 8.63 (d, J = 2.02 Hz, 1H), 8.50-8.60 (m, 1H) J = 9.09 Hz, 1H), 7.16 (d, J = 9.35 Hz, 1H), 6.88 (br s, 1H), 3.44 (q, J = 6.57 Hz, 2H), 3.38 (br s, 1H), 2.98 (d, J = 6.06 Hz, 2H), 1.61 (d, J = 6.57 Hz, 2H), 1, 43-1.54 (m, 2H), 1.39 (s, 9H); LCMS (LCMS Method C): retention time = 0.91 min, [M + H] + = 354, 1.

Stage 2:

tert-Butyl (4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) carbamate

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To 4 - ((4 - ((tert-butoxycarbonyl) amino) butyl) amino) -3-nitrobenzoic acid hydrochloride (10.7 g, 27.4 mmol) and HATU (12.5 g, 32.9 mmol) in DCM (91 ml) was added DIEA (10.5 ml, 60.3 mmol) at room temperature, followed by the addition of 7M ammonia in MeOH (7.83 ml, 54.8 mmol). After 3 hours, the resulting yellow solid was collected by filtration and washed with DCM to give the title compound (8.52 g, 21.8 mmol, 79% yield). 1H NMR (400 MHz, DMSOX ₆ ) δ ppm 8.41 (t, J = 5.68 Hz, 1H), 8.34 (dd, J = 4.04, 1.26 Hz, 1H), 8.13 (dd, J = 8.34, 1.26 Hz, 1H), 7.31 (br s, 2H), 7.07-7.15 (m, 1H), 3.39-3.46 (m , 2H), 3.36 (br. C, 1H), 2.97, J = 6.57 Hz, 2H), 1.55-1.67 (m, 2H), 1.42-1.53 ( m, 2H), 1.38 (s, 9H); LCMS (LCMS Method C): retention time = 0.8 4 min, [M + H] + = 353.1.

Stage 3:

4 - ((4-aminobutyl) amino) -3-nitrobenzamide dihydrochloride

A 4 M solution of HCl in dioxane ( 62.8 ml, 251 mmol). After 2 to 4 hours, the reaction mixture was concentrated to afford the title compound (4.08 g, 11.9 mmol, 95% yield) as a yellow-orange solid. 1H NMR (400 MHz, DMSOX ₆ ) δ ppm 8.78 (dd, J = 4.42, 1.39 Hz, 1H), 8.68 (d, J = 2.02 Hz, 1H), 8.55 (dd, J = 8.34, 1.52 Hz, 1 H), 8.03 (dd, J = 9.09, 2.02 Hz, 2H), 7.90 (br s, 3H), 7 , 09-7.17 (m, 2H), 3.46 (d, J = 6.06 Hz, 2H), 2.78-2.91 (m, 2H), 1.67 (br C, 4H ); LCMS (LCMS Method C): retention time = 0.40 min, [M + H] ⁺ = 253.0.

Stage 4:

Methyl 4 - ((4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) amino) -3-nitrobenzoate

To a suspension of 4 - ((4-aminobutyl) amino) -3-nitrobenzamide dihydrochloride (1.84 g, 5.66 mmol) and K2CO ₃ (2.346 g, 16.97 mmol) in DMSO (11.32 ml) at 23 ° C methyl 4-fluoro-3-nitrobenzoate (1.13 g, 5.66 mmol) was added. After 30 min, a bright yellow precipitate formed. The reaction mixture was diluted with water (25 ml) and the solid was collected by filtration and dried in vacuo to give the title compound (4.1 g, 5.6 mmol, 99% yield). ¹ H NMR (400 MHz, DMSOX ₆ ) δ ppm 8.64 (d, J = 2.28 Hz, 1H), 8.61 (d, J = 2.28 Hz, 2H), 8.42 (t, J = 5.70 Hz, 1H), 7.93-8.03 (m, 3H), 7.30 (br s, 1H), 7.17 (d, J = 9.38 Hz, 1H), 7.12 (d, J = 9.12 Hz, 1H), 3.83 (s, 3H), 3.48 (d, 6.08 Hz, 4H), 1.73 (bs, 4H); LCMS (LCMS Method D): retention time = 0.97 min, [M + H] + = 432.2.

Stage 5:

Methyl 2-amino-1 - (4- (2-amino-5-carbamoyl-1 H-benzo ^] imidazol-1-yl) butyl) -1 H-benzoI imidazole 5-carboxylate dihydrobromide

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K methyl 4 - ((4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) amino) -3-nitrobenzoate (3.18 g, 7.37 mmol) and 10% Pd / C (1.2 g , 1.12 8 mmol) at room temperature was added NMP (35 ml). The flask was evacuated and filled with nitrogen then re-evacuated and charged with 1 atm of H ₂ fed from a cylinder. The mixture was stirred at 70 ° C for 18 h, then filtered through celite. The filtrate was cooled to room temperature and treated with cyanogen bromide (2.95 mL, 14.7 mmol). After 2 h, the reaction mixture was heated to 70 ° C for 2 h, cooled to room temperature and diluted with EtOAc (120 ml) with vigorous stirring. The resulting gray solid was collected by filtration to give the title compound (4.91 g, 5.90 mmol, 80% yield). ¹ H NMR (400 MHz, D \ 1CO) -s1 „) δ ppm 12.85 (brs, 2H), 8.91 (brs, 2H), 8.78 (s, 2H), 8, 08 (bs, 1H), 7.82-7.99 (m, 4H), 7.60-7.75 (m, 2H), 7.47 (bs, 1H), 4.13- 4.28 (m, 4H), 3.89 (s, 3H), 1.74-1.84 (m, 4H); LCMS (LCMS Method C): retention time = 0.49 min, [M + H] + = 422.2.

Example 23.

Stage 6:

Methyl 1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo | | imidazol-1 yl) butyl) -2- (1-ethyl -3 -methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo [d] imidazole-5-carboxylate-bistrifluoroacetate

Contents of a microwave reactor vial with 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (0.502 g, 3.26 mmol), HATU (1.27 g, 3.34 mmol) and DIEA (1.497 ml, 8.57 mmol) in NMP (5.71 ml) was stirred at room temperature for 10 min. A separate solution of methyl 2amino-1 - (4- (2-amino-5-carbamoyl-1H-benzoOl | imidazol-1-yl) butyl) -1H-benzo ^ | imidazole-5-carboxylate dihydrobromide (1.0 g , 1.7 mmol), dissolved in a minimum amount of NMP, the vial was sealed and heated in a microwave reactor to 140 ° C. After 30 min, the reaction mixture was cooled to room temperature and diluted with water (about 50 ml). The resulting gray solid was collected by filtration, dissolved in the minimum amount of DMSO and purified by reverse phase HPLC (Luna C18 50x30 mm column, 47 ml / min) eluting with 10-40% acetonitrile in water (0.1% TFA) to give the title compound (400 mg, 0.412 mmol, 24% yield) as an off-white solid. 1H NMR (400 MHz, DMSO-dg) δ ppm 12.89 (s, 1H), 12.83 (s, 1H), 8.09 (s, 1H), 7.93-8.01 (m, 2H ), 7.84 (d, J = 8.59 Hz, 1H), 7.77 (d, J = 8.08 Hz, 1H), 7.58 (dd, J = 17.68, 8.34 Hz , 2H), 7.35 (br s, 1H) (m, 4H), 3.88 1.82-1.94 (m, 4H), 1.31 (t, J = 6.82 Hz, 6H ); LCMS (LCMS Method C): retention time = 0.86 min, [M + H] + = 694.6.

Example 24.

(E) -1,1 '- (but-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1Nbenzo [ d] imidazole-5-carboxamide)

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Example 24 can be prepared according to method 1 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

A solution of 1-allyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1 H-benzo [d] imidazole 5-carboxamide (20 mg, 0.052 mmol) in 1.2 -dichloroethane (2.1 ml) was degassed with N2 at 90 ° C (heating is necessary for dissolution) for 5 min, then Zhang IB's catalyst (CAS 918870-76-5, 5.76 mg, 7.84 μmol) was added. After heating for 18 hours, the reaction mixture was stirred at room temperature for 2 days. An additional 10 mg of liquid Zhang catalyst 1B was added and the reaction mixture was reheated to 90 ° C. After 1 hour, the mixture was cooled, and the resulting solid was collected by filtration to give the title compound (4 mg, 5 μmol, 10% yield). ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.84 (s, 2H), 7.91-8.24 (m, 2H), 7.64 (s, 2H), 7.33 (s, 4H), 6.53 (s, 2H), 5.70-6.16 (m, 2H), 4.91 (bs, 4H), 4.29-4.44 (m, 4H), 3.77 (s, 6H), 2.10 (s, 6H), 1.27 (s, 6H); LCMS (LCMS Method L): retention time = 0.85 min, [M + H] + = 737.5.

Example 25.1,1 '- (Butane-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -7 - (3-hydroxypropoxy) -1H-benzoSimidazole -5-carboxamide) dihydrochloride

Example 25 can be prepared according to method 1 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

K 1,1 '- (butane-1,4-diyl) bis (7- (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide) (12 mg, 0.011 mmol) in 1,4-dioxane (1.5 ml) was added a 4N solution of HCl in dioxane (0.011 ml, 0.045 mmol). After 60 min, the reaction mixture was concentrated and triturated with EtOAc, then the solid was isolated by filtration and dried to give the title compound (10 mg, 0.011 mmol, 98% yield) as a white solid. 1H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.77 (brs, 2H), 8.02 (brs, 2H), 7.57 (s, 2H), 7.297.39 (m, 4H ), 6.58 (s, 2H), 4.56 (d, J = 7.10 Hz, 4H), 4.36 (br s, 4H), 4.15 (t, J = 6.21 Hz , 4H), 3.48-3.54 (m, 6H), 2.10 (s, 6H), 1.86 (br s, 4H), 1.74-1.83 (m, 4H), 1.30 (t, J = 7.10 Hz, 6H); LCMS (LCMS Method D): retention time = 0.78 min, [M + H] + = 827.4.

Example 26 (E) -8-ethyl-1,26-bis (3-hydroxypropoxy) -10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15, 20,21,28,31-dodecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3' 1] [1,3,6,15,17] pentaazacycloneicosin-3,2 4-dicarboxamide-bis-hydrochloride

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Example 26 can be prepared according to method 1 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

K (E) -4,26-bis (3 - ((tert-Butyldimethylsilyl) oxy) propoxy) -8-ethyl-10,18-dimethyl-7,20-dioxo6,7,8,11,12,13, 14,15,20,21,28,31-dodecahydrorbenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] diparazolo [5,1-e: 4 ' , 3'-1] [1,3,6,15,17] pentaazacyclogeneicosine-2,24-dicarboxamide (269 mg, 0.246 mmol) in THF (20 ml) at 0 ° C was added dropwise 4 N HCl solution in dioxane (0.31 ml, 1.23 mmol). After 2 h, the resulting white solid was collected by filtration and washed with Et ₂ O to give the title compound (226 mg, 0.241 mmol, 98% yield). 1H NMR (600 MHz, MeOH-d ₄ ) δ ppm 7.71-7.74 (m, 2H), 7.38-7.41 (m, 2H), 6.82 (s, 1H), 5, 72 (s, 2H) 5.07 (m, 4H), 4.77-4.83 (m, 2H), 4.67 (q, J = 7.2 Hz, 2H), 3.88-3, 98 (m, 4H), 3.39-3.34 (m, 4H), 2.85-2.92 (m, 2H), 2.41 (s, 3H), 2.38 (s, 3H) , 1.93-2.02 (m, 2H), 1.67 (t, J = 7.6 Hz, 2 H), 1.49 (t, J = 7.2 Hz, 5H), 1.34 -1.44 (m, 4H); LC-MS (LC-MS Method L): retention time ^ ,? 3 min, [M + H] + = 865.7017.

Example 27. (E) -1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzoShimidazole-1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7 (3-methoxypropoxy) -1H-benzoHimidazole-5-carboxamide, dihydrochloride

Example 27 can be prepared according to a combination of methods 2, 3 and 4, with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (216 mg, 1.40 mmol) and CDI (22 7 mg, 1.40 mmol) were stirred in DMF (0.7 ml) at 100 ° С ... After 10 min, (E) -2-amino-1- (4- (2-amino-5carbamoyl-7- (3-hydroxypropoxy) -1H-benzoShimidazol-1-yl) but-2-1-yl) -7 - (3-methoxypropoxy) -1N-benzo-imidazole-5-carboxamide dihydrobromide (255 mg, 0.350 mmol) and the reaction mixture was heated at 110 ° C. After 18 h, 10 M NaOH solution (350 μL, 3.50 mmol) was added and the reaction mixture was heated for 1 h at 75 ° C to hydrolyze a small amount of ester by-product (15%). The mixture was then poured into saturated aqueous NH4C1 (10 ml) and the resulting solid was collected by filtration. The solid was suspended in hot MeOH and filtered to give a white powder. This solid was suspended in dioxane (10 ml) and 4N HCl in dioxane (74 μl, 0.30 mmol) was added. After 10 min, the solid was collected by filtration and washed with diethyl ether to give the title compound (110 mg, 0.121 mmol, 35% yield). 1H NMR (400 MHz, MeOH-d4) δ ppm 7.66 (dd, J = 5.32, 1.01 Hz, 2H), 7.39 (dd, J = 13.18, 1.27 Hz, 2H ), 6.68 (d, J = 17.24 Hz, 2H), 5.93 (br s, 2H), 5.17 (br s, 4H), 4.61-4.75 (m, 4H), 4.06-4.22 (m, 4H), 3.613.72 (m, 2H), 3.45 (t, J = 6.08 Hz, 2H), 3.31 (s, 3H), 2.28 (d, J = 4.06 Hz, 6H), 1.88 (td, J = 6.21, 2.28 Hz, 4H), 1.35-1.50 (m, 6H); LCMS (LCMS method L): retention time = 0.78 min, [M + H] + = 839.6204.

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Example 28. (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (2-methoxyethoxy) -1H-benzo

Shimidazole-5-carboxamide

Example 28 can be prepared according to a combination of methods 2 and 3 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

A mixture of HATU (894 mg, 2.35 mmol), DIEA (425 mg, 3.29 mmol), (E) -2-amino-1- (4- (2-amino-5carbamoyl-1 H-benzo [d] imidazol-1-yl) no-2-en-1-yl) -7- (2-methoxyethoxy) -1H-benzo [d] imidazole-5-carboxamide (450 mg, 0.940 mmol) and 1-ethyl-3- methyl 1H-pyrazole-5-carboxylic acid (362 mg, 2.35 mmol) in DMF (8 ml) was stirred at 65 ° C. After 12 hours, the reaction mixture was cooled and treated with water. The resulting solid was collected by filtration and purified by HPLC to give the title compound (350 mg, 0.466 mmol, 50% yield). 1H NMR (400 MHz, DMSOh! ₆ ) δ ppm 12.85 (s, 2H), 7.95-7.99 (m, 4H), 7.65-7.67 (m, 1H), 7.60 (s, 1H) 7.48 (m, 1H), 7.28-7.35 (m, 2H), 6.55 (d, J = 4.0 Hz, 2H), 5.99 (dd, J = 13.2, 7.7 Hz, 1H), 5.85 (dd, J = 13.3, 7.5 Hz, 1H), 4.94 (d, J = 5.3 Hz, 2H), 4 , 83 (d, J = 4.9 Hz, 2H), 4.58-4.49 (m, 4H), 4.15-4.12 (m, 2H), 3.58-3.49 (m , 2H), 3.16 (s, 3H), 2.12 (s, 6H), 1.26-1.35 (m, 6H); LCMS (LCMS Method A): retention time = 1.353 min, [M + H] + = 751.1.

Example 29 (E) -1,1 '- (But-2-en-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-isopropoxy -1 N-benzo Schimidazole-5-carboxamide)

Example 29 can be prepared according to method 2 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (89 mg, 0.58 mmol) and CDI (107 mg, 0.659 mmol) in DMF (2 ml) were stirred for 10 min. (E) -1,1 '- (but-2-ene-1,4-diyl) bis (2-amino-7isopropoxy-III-benzoHimidazole-5-carboxamide) dihydrobromide (180 mg, 0.244 mmol) and DIEA ( 0.18 ml, 1.1 mmol) and the reaction mixture was heated at 90 ° C. After 3 h, ice was added and the resulting solid was collected by filtration, stirred vigorously for 3 h in EtOAc and isolated. The solid contained about 5% monoamide by LC-MS and was treated with CDI (14 mg, 0.086 mmol), 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (12 mg, 0.078 mmol) and DIEA (0.1 ml, 0.6 mmol) in DMF (1.5 ml). The reaction mixture was heated to 90 ° C for 2 h and another portion of reagents was added. After 4 h, the reaction mixture was cooled to room temperature, ice was added and the resulting solid was collected by filtration to give the title compound (122 mg, 0.154 mmol, 58% yield). ¹ H NMR (400 MHz, DMSO-d) δ ppm 12.80 (s, 2H), 7.94 (brs, 2H), 7.59 (s, 2H), 7.32 (s, 2H) , 2H), 6.55 (s, 2H), 5.85 (br s, 2H), 4.94 (br s, 4H), 4.74 (dt, J = 12.0, 5.8 Hz, 2H) 4.75 (d, J = 7.0 Hz, 4H), 2.14 (s, 6H), 1.28 (t, J = 7.0 Hz, 6H), 1.10 (d , J = 6.1 Hz, 12H); LCMS (LCMS Method D): retention time = 1.03 min, [M / 2 + H] + = 397, 4557.

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Example 30 (E) -7- (Benzyloxy) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 7-methoxy-1H-benzo ^ ] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo ^] imidazole-5-carboxamide

Example 30 can be prepared according to a combination of methods 2, 3 and 4, with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (20.2 mg, 0.131 mmol) and CDI (23.1 mg, 0.142 mmol) in DMF (1 ml) were stirred for 10 min. (E) -2-amino-1- (4- (2-amino-5carbamoyl-7-methoxy-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7- ( benzyloxy) -1H-benzo (^] imidazole-5carboxamide dihydrobromide (40 mg, 0.057 mmol) and DIEA (0.07 ml, 0.4 mmol) and the reaction mixture was heated at 90 ° C. After 22 h, ice was added with vigorous stirring and the resulting solid was collected by filtration, washed with water and triturated with ethyl acetate and methanol successively to give the title compound (10 mg, 0.012 mmol, 21% yield) as a yellow brown solid. ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.86 (d, J = 5.8 Hz, 2H), 7.99 (brs, 2H), 7.67 (d, J = 2, 3 Hz, 2H) (s, 1H), 7.37 (brs, 2H), 7.19-7.30 (m, 6H), 6.53 (s, 1H), 6.49 (s, 1H), 5.74-5.84 (m, 1H), 5.53-5.62 (m, 1H), 5.05 (s, 2H), 4.86 (dd, J = 11.8 , 4.9 Hz, 4H), 4.44-4.58 (m, 4H), 3.64 (s, 3H), 2.12 (s, 3H), 2.09 (s, 3H), 1 , 19-1.31 (m, 6H); LCMS (LCMS Method D): retention time = 1.02 min, [M / 2 + H] + = 407.4811.

Example 31. (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazol-1-yl) but- 2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methyl-1 H-benzoI imidazole 5-carboxamide

Example 31 can be prepared according to a combination of methods 2 and 4 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

A mixture of HATU (568 mg, 1.49 mmol), DIEA (0.31 ml, 1.8 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5 carboxylic acid (203 mg, 1.31 mmol) in NMP (5 ml) was stirred at room temperature. After 1 h, (E) -2-amino-1- (4- (2-amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7 methyl -1H-benzo ^] imidazole-5-carboxamide (250 mg, 0.597 mmol) and the mixture was stirred at 60 ° C overnight. The reaction mixture was treated with water and the resulting solid was collected by filtration and purified by HPLC (Gemini-C18 column, 150x21.2 mm, 5 μm, 20-50% CH ₃ CN in H ₂ O, 0.1% TFA) to give the title compounds (3 mg, 4 μmol, yield 0.7%). 1H NMR (400 MHz, DMSO d6) δ ppm 13.08-12.70 (m, 2H), 7.97 (d, J = 10.4 Hz, 2H), 7.87 (d, J = 13, 0 Hz, 2H) d, J = 8.6 Hz, 1H), 7.50 (s, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.32 (d, J = 17.7 Hz, 2H), 6.54 (d, J = 7.9 Hz, 2H), 5.96 (d, J = 15.3 Hz, 1H), 5.52 (d, J = 15, 6 Hz, 1H), 4.99 (s, 2H), 4.83 (s, 2H) 4.53 (d, J = 4.7 Hz, 4H), 2.51 (s, 3H), 2, 12 (d, J = 2.8 Hz, 6H), 1.27 (t, J = 7.1 Hz, 6H); LCMS (LCMS Method A): retention time = 1.321 min, [M + H] + = 691.3.

Example 32 (E) -1,1 '- (But-2-en-1,4-diyl) bis (7-butoxy-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1 N-benzo ^] imidazole-5-carboxamide)

- 100 036435

Example 32 can be prepared according to method 2 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

To 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (17.4 mg, 0.113 mmol) and CDI (18.3 mg, 0.113 mmol) in DM FA (650 μL) at 60 ° C was added ( E) -1,1 '- (but-2-en-1,4-diyl) bis (2-amino-7- (3-hydroxypropoxy) -1H-benzo ^] imidazole-5-carboxamide) (25 mg, 0.045 mmol ) and TEA (38 μl, 0.27 mmol) and the reaction mixture was heated to 120 ° C. After 18 h, an additional amount of CDI solution (30 mg, 0.19 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (30 mg, 0.19 mmol) were added (heated in DMF for 10 min at 60 ° C). The reaction mixture was cooled, water was added and the resulting precipitate was collected by filtration and triturated with hot MeOH (1 ml) to give the title compound (6.9 mg, 8.4 μmol, 13% yield) as a tan solid. ¹ H

NMR (400 MHz, DMSO ^ b) δ ppm 12.87 (s, 2H), 7.96 (brs, 2H), 7.65 (s, 2H), 7.35 (brs, 7, 26 (s, 2H), 6.56 (s, 2H), 5.81 (bs, 2H), 4.91 (bs, 4H), 4.49-4.42 (m, 4H) , 3.84 (t, J = 6.46 Hz, 4H), 2.14 (s, 6H), 1.37-1.48 (m, 5H), 1.30 (t, J = 6.97 Hz, 6H), 1.20 (dd, J = 14.95, 7.60 Hz, 4H), 0.74 (t, J = 7.48 Hz, 6H); LC-MS (LC-MS method L ): retention time = 1.0 6 min, [M + H] + = 821, 7039.

Example 3 3. (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^] imidazol-1-yl ) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-isopropoxy-1 H-benzo ^] imidazole-5-carboxamide

Example 33 can be prepared according to a combination of methods 2 and 4 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

A mixture of HATU (190 mg, 0.500 mmol), DIEA (0.16 ml, 0.93 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5 carboxylic acid (70 mg, 0.45 mmol) in NMP (3 ml) was stirred at room temperature. After 15 min, (E) -2-amino-1- (4- (2-amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7 isopropoxy -1H-benzo ^] imidazole-5-carboxamide (86 mg, 0.19 mmol) and the mixture was stirred at 60 ° C for 16 h. The reaction mixture was treated with water and the resulting solid was collected by filtration and then purified by HPLC (Gemini column -C18, 150x21.2 mm, 5 μm, 30-50% CH3CN in H ₂ O, 0.1% TFA, 15 min) to give the title compound (23 mg, 0.031 mmol, 17% yield) as an off-white solid. 1H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.82 (s, 2H), 7.96 (d, J = 14.3 Hz, 3H), 7.72 (d, J = 8.5 Hz, 1H ), 7.61 (s, 1H), 7.44 (d, J = 8.4 Hz, 1H), 7.31 (d, J = 11.6 Hz, 3H), 6.55 (s, 2H ), 5.96 (dd, J = 13.5, 7.7 Hz, 1 H), 5.80 (d, J = 15.7 Hz, 1H), 4.94 (d, J = 5.4 Hz, 2H), 4.84 (d, J = 4.9 Hz, 2H), 4.73-4.79 (m, 1H), 4.54 (td, J = 14.3, 7.1 Hz , 4H), 2.13 (d, J = 5.8 Hz, 6H), 1.28 (dt, J = 12.1, 7.1 Hz, 6H), 1.14 (d, J = 6, 0 Hz, 6H); LCMS (LCMS Method A): retention time = 1.413 min, [M + H] + = 735.2.

Example 34. (E) -1,1 '- (but-2-ene-1,4-diyl) bis (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- ( 3isopropoxypropoxy) -1H-benzoI imidazole-5-carboxamide) dihydrochloride

- 101 036435

Example 34 can be prepared according to method 2 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (110 mg, 0.711 mmol), HATU (271 mg, 0.711 mmol) and HOBt (54.5 mg, 0.35 6 mmol) in DMF (0.9 ml) suspension (E) -1,1 '- (but-2-en1,4-diyl) bis (2-amino-7- (3-isopropoxypropoxy) -1 H-benzo ^] - imidazole was added -5-carboxamide) (151 mg, 0.377 mmol) and TEA (0.20 ml, 1.4 mmol) in DMF (3.8 ml). After stirring at room temperature overnight, the reaction mixture was diluted with water, extracted with EtOAc (2x), and the combined organic extracts were washed with brine and concentrated. The resulting residue was purified on silica gel (40 g Isco column) eluting with 0-20% MeOH in DCM to give the free base of the title compound (105 mg, 0.116 mmol, 49% yield) as a pale yellow solid. A portion of this substance (80 mg, 0.088 mmol) was dissolved in MeOH (2 ml) and DCM (2 ml) and treated with 4N HCl solution in dioxane (0.044 ml, 0.18 mmol). After 5 min, the reaction mixture was added to MeCN (very little solid precipitated) and the mixture was concentrated. The resulting residue was triturated with EtOAc to give the title compound (68 mg, 0.069 mmol, 79% yield) as a white solid. ¹ H NMR (400 MHz, DMSO-dg) δ ppm 8.00 (brs, 2H), 7.65 (d, J = 1.01 Hz, 2H), 7.35 (brs, 2H) , 7.30 (d, J = 1.01 Hz, 2H), 6.55 (s, 2H), 5.83 (br s, 2H), 4.94 (br s, 4H), 4, 54 (q, J = 7.10 Hz, 4 H), 4.00 (t, J = 6.21 Hz, 4H), 3.32-3.41 (m, 2H), 3.21-3, 31 (m, 4H), 2.13 (s, 6H), 1.69 (t, J = 6.21 Hz, 4H), 1.29 (t, J = 7.10 Hz, 6H), 0, 98 (d, J = 6.08 Hz, 12H); LCMS (LCMS Method D): retention time = 1.10 min, [M / 2 + H] + = 455.5857.

Example 3 5. (E) -2- (1-Ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1 - (4- (2- (1-ethyl-3-methyl-1H-pyrazole- 5-carboxamido) -7- (morpholinomethyl) -1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -1H-benzo ^] imidazole-5-carboxamide trifluoroacetate

Example 35 can be prepared according to a combination of methods 2 and 3 with modifications known to one of ordinary skill in the art.

The last stage of obtaining is given:

A mixture of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (117 mg, 0.760 mmol), HATU (347 mg, 0.912 mmol) and DIEA (0.319 ml, 1.824 mmol) in DMF (6 ml) was stirred at room temperature. After 30 min, (E) -2-amino-1- (4- (2-amino-7- (morpholinomethyl) -1H-benzo ^] imidazol-1-yl) but2-en-1-yl) -1H- benzo ^] imidazole-5-carboxamide (140 mg, 0.304 mmol) and the reaction mixture was stirred at 50 ° C overnight. Water was added and the resulting solid was collected by filtration and purified by preparative HPLC to give the title compound (10 mg, 12 μmol, 3.9% yield) as a gray solid. 1H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.98 (br s, 2H), 10.59 (br s, 1H), 7.91-8.01 (m, 2H), 7.74 (d, 8.4 Hz, 1H), 7.64 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.247.39 (m, 3H ), 6, 56 (d, J = 5.6 Hz, 2H), 5.93 (d, J = 16 Hz, 2H), 5.38-5.50 (m, 2H), 5.10 (br s, 2H), 4.79 (br s, 2H), 4.48-4.55 (m, 4H), 3.69-3.85 (m, 4H), 3.03-3.32 (m, 4H), 2.13 (s, 6H), 1.21-1.40 (m, 6H); LCMS (LCMS Method A): retention time = 1.328 min, [M + H] ⁺ = 733.2.

Example 36 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (piperidin4-yl) propoxy) - 1 H-benzoI imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxa-102 036435 mido) -7-methoxy- 1Н-benzo ^] imidazole-5-carboxamide, n hydrochloride

HCI, MeOH room temp.

DIAD, RRpZ

THF, room temp.

Na ₂ S ₂ O ₄ , NH4OH

MeON, N ₂ O room temp.

EDC, TEA, room temp.

Stage 1:

tert-Butyl 4- (3- (5-carbamoyl-2-chloro-3-nitrophenoxy) propyl) piperidine-1-carboxylate

4-Chloro-3-hydroxy-5-nitrobenzamide (1 g, 4.62 mmol), tert-butyl 4- (3-hydroxypropyl) piperidine 1-carboxylate (1.348 g, 5.54 mmol) and triphenylphosphine (2.059 g, 7 , 85 mmol) was mixed in THF (20 ml) at 0 ° C and then diisopropyl (E) -diazole-1,2-dicarboxylate (1.54 5 ml, 7.85 mmol) was added. The reaction was maintained at room temperature for 16 h, then the yellow solution was concentrated and the residue was partitioned between aqueous NaHCO ₃ and EtOAc. The organic layer was washed with brine, dried over MgSO ₄ and concentrated to a yellow residue. This residue was purified by Isco Combiflash (10% -50% (3: 1 EtOAc / EtOH) / hexane, with 2% NH4OH, 40 g RediSep column). The collected fractions containing the product were combined and concentrated to give the title compound as a yellow solid (2.83 g, 61% purity, 3.91 mmol, 85% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm 8.29 (s, 1H), 8.04 (d, J = 1.77 Hz, 1H), 7.87 (d, J = 1.52 Hz, 1H), 7.80 (s, 1H), 4.22 (t, J = 6.34 Hz, 2H), 3.93 (d, J = 10.90 Hz, 2H), 2.68 (br. s, 2H), 1.77-1.88 (m, 2H), 1.67 (d, J = 11.41 Hz, 2H), 1.47 (br s, 1H), 1.34-1 , 42 (m, 11H), 0.89-1.05 (m, 2H); LCMS (LCMS Method K): retention time = 1.2-5 min, [M-100] ⁺ = 342.0.

Stage 2:

tert-Butyl (E) -4- (3- (5-carbamoyl-2 - ((4 - ((4-carbamoyl-2-methoxy-6-nitrophenyl) amino) but-2-en-1yl) amino) - 3-nitrophenoxy) propyl) piperidine-1-carboxylate

- 103 036435

(E) -4 - ((4-Aminobut-2-en-1-yl) amino) -3-methoxy-5-nitrobenzamide trifluoroacetate (1 g, 2.54 mmol) was suspended in n-butanol (10 ml) at room temperature, then DIPEA (2.66 ml, 15.22 mmol) and tert-butyl 4- (3- (5-carbamoyl-2-chloro-3-nitrophenoxy) propyl) piperidine-1-carboxylate (2.021 g, 2.79 mmol). Then the reaction mixture was kept at 120 ° С for 48 h, then the reaction mixture was cooled to room temperature. The brown solid was collected by filtration and purified by Isco Combiflash (20% -80% (3: 1 EtOAc / EtOH) / hexane with 2% NH ₄ OH, RediSep 40 g column). The collected fractions containing the product were combined and concentrated to give the title compound as a red solid (204 mg, 12% yield). LCMS (LCMS Method K): retention time = 1.2 5 min, [M-100] ⁺ = 586.2.

Stage 3:

tert-Butyl (E) -4- (3- (3-amino-2 - ((4 - ((2-amino-4-carbamoyl-6-methoxyphenyl) amino) but-2-en-1yl) amino) - 5-carbamoylphenoxy) propyl) piperidine-1-carboxylate

The sodium salt of hydrosulfurous acid (609 mg, 2.97 mmol) was dissolved in H ₂ O (5 ml) at room temperature and then this solution was added to a stirred solution of tert-butyl (E) -4- (3- (5carbamoyl-2- ((4 - ((4-carbamoyl-2-methoxy-6-nitrophenyl) amino) but-2-en-1-yl) amino) -3-nitrophenoxy) propyl) piperidine-1-carboxylate (204 mg, 0.297 mmol ) and ammonium hydroxide (0.799 ml, 5.95 mmol) in 20 ml of MeOH. Then the reaction mixture was kept at room temperature for 2 h, then the mixture was filtered and the filtrate was partially concentrated to remove MeOH. The resulting yellow aqueous mixture was partitioned between saturated NaHCO ₃ (aq) and EtOAc. The organic layer was washed with brine, dried over MgSO ₄ , concentrated and purified by Isco Combiflash (2% -20% MeOH / CHr ^ g, 10% TEA in MeOH, 40 g RediSep column).

The collected fractions containing the product were combined and concentrated to give the title compound as a colorless oil (77 mg, 42% yield). ¹ H NMR (400 MHz, methanol ^ ₄ ) δ ppm 6.94 (t, J = 2.15 Hz, 2H), 6.89 (dd, J = 4.56, 1.77 Hz, 2H), 5 , 74 (d, J = 4.56 Hz, 2 H), 4.05 (d, J = 13.43 Hz, 2H), 3.96 (t, J = 6.34 Hz, 2H), 3, 80 (s, 3H), 3.52-3.62 (m, 4H), 2.62-2.83 (m, 2H), 1.77-1.88 (m, 2H), 1.71 ( d, J = 11.91 Hz, 2H), 1.35-1.52 (m, 12H), 0.99-1.16 (m, 2H). LCMS (LCMS Method K): retention time = 0.64 min, [M + H] ⁺ = 626.3.

Stage 4:

tert-Butyl (E) -4- (3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- methoxy-1 H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) 1 H-benzo ^] imidazol-7-yl) oxy) propyl) piperidine-1 -carboxylate

- 104 036435 tert-Butyl E-4- (3- (3-amino-2 - ((4 - ((2-amino-4-carbamoyl-6-methoxyphenyl) amino) but-2-en-1-yl) amino) -5-carbamoylphenoxy) propyl) piperidine-1-carboxylate (77 mg, 0.123 mmol) was dissolved in DMF (3 ml) at 0 ° C and then 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate was added (0.308 ml, 0.123 mmol). Then the reaction mixture was kept at 0 ° C for 15 min, then EDC (28.3 mg, 0.148 mmol) and TEA (0.043 ml, 0.308 mmol) were added to the reaction mixture. The reaction mixture was then kept at room temperature for 16 hours. The reaction mixture was concentrated and the yellow residue was purified on silica gel (20-50% MeOH / CH ₂ Cl ₂ , 10% TEA in MeOH, 40 g RediSep column). The collected fractions containing the product were combined and concentrated to give the title compound as a white solid (87 mg, 52% yield). LCMS (LCMS Method K): retention time = 1.11 min, [M + H] + = 948.3.

Example 36 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (piperidin4-yl) propoxy) - 1 H-benzo (imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -7-methoxy-1 H- benzo [d] imidazole-5-carboxamide hydrochloride

Stage 5:

tert-Butyl (E) -4- (3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- methoxy-1 H-benzo (imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) 1H-benzoShimidazole-7- yl) oxy) propyl) piperidine-1-carboxylate (87 mg, 0.0 92 mmol) was suspended in MeOH (40 ml) and HCl (4N solution in 1,4-dioxane) (0.575 ml, 2.30 mmol ). The reaction mixture was kept at room temperature for 48 h, then the mixture was concentrated and the residue was purified by HPLC (XSELECT CSH C18 column, 150 mm × 30 mm, inner diameter 5 μm, a mixture of 15% -85% 10 mM ammonium bicarbonate solution in water with acetonitrile). The desired fractions were combined and partially concentrated in vacuo and the white precipitate was collected to give the title compound as a white solid (30 mg, 37% yield). ¹ H NMR (400 MHz, methanol ^ ₄ ) δ ppm 8.57 (s, 1H), 7.63 (d, J = 1.01 Hz, 1H), 7.57 (s, 1H), 7.29 (s, 1H) (s, 1H), 6, 67 (s, 1H), 6.58 (s, 1H), 5.85 (bs, 2H), 5.03 (bs, 4H) , 4.54-4.73 (m, 4H), 3.81 (t, J = 6.46 Hz, 2H), 3.71 (s, 3H), 3.23 (d, J = 12.93 Hz, 2H), 2.72-2.85 (m, 2H), 2.24 (d, J = 12.17 Hz, 6H), 1.74 (d, J = 13.69 Hz, 2H), 1.12-1.54 (m, 13H). LCMS (LCMS Method K): retention time = 0.72 min, [M + H] + = 848.6.

Example 37 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (piperazin1-yl) propoxy) - 1 H-benzo (imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -7-methoxy-1 H- benzo U | imidazole-5-carboxamide

- 105 036435

Stage 1:

tert-Butyl 4- (3- (5-carbamoyl-2-chloro-3-nitrophenoxy) propyl) piperazine-1-carboxylate

Triphenylphosphine (2.059 g, 7.85 mmol), tert-butyl 4- (3-hydroxypropyl) piperazine-1-carboxylate (1.692 g, 6.93 mmol) and diisopropyl (E) -diazen-1,2-dicarboxylate (1.587 g, 7.85 mmol) was mixed in THF (20 ml) at 0 ° C and then 4-chloro-3-hydroxy-5-nitrobenzamide (1 g, 4.62 mmol) was added. The reaction solution was kept at room temperature for 16 h, then the brown reaction solution was partitioned between saturated NaHCO ₃ (aq) and EtOAc. The organic layer was washed with brine, dried over MgSO ₄ , concentrated and purified on silica gel (20% -80% (3: 1 EtOAc / EtOH) / hexane, with 2% NH4OH, 330 g RediSep column). The desired fractions were combined and concentrated to give the title compound as a white solid (970 mg, 47% yield). ¹ H NMR (400 MHz, DMSO-06) δ ppm 8.30 (s, 1H), 8.05 (d, J = 1.77 Hz, 1H), 7.88 (d, J = 1.77 Hz , 1H), 7.80 (s, 1H), 4.28 (t, J = 6.21 Hz, 2H), 3.31 (brs, 4H), 2.48 (t, J = 7, 10 Hz, 2H), 2.33 (t, J = 4.94 Hz, 4H), 1.96 (t, J = 6.59 Hz, 2H), 1.40 (s, 9H). LCMS (LCMS Method K): retention time = 0.69 min, [M + H] + = 443.4.

Stage 2:

tert-Butyl (E) -4- (3- (5-carbamoyl-2 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- methoxy-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) amino) -3-nitrophenoxy) propyl) piperazine-1carboxylate

- 106 036435

(E) -1 - (4-Aminobut-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1Hbenzo ^] imidazole- 5-carboxamide dihydrochloride (242 mg, 0.499 mmol) was dissolved in n-butanol (10 ml) at room temperature, and then DIPEA (0.476 ml, 2.72 mmol) was added followed by tert-butyl 4- (3- (5 -carbamoyl-2-chloro-3-nitrophenoxy) propyl) piperazine-1-carboxylate (201 mg, 0.454 mmol). The reaction mixture was kept at 120 ° C for 16 h. The reaction mixture was cooled to room temperature and the red solid was collected by filtration (296 mg, 73% yield). ¹ H NMR (400 MHz, DMSO-06) δ ppm 8.14 (d, J = 1.77 Hz, 1H), 8.00 (brs, 2H), 7.84 (t, J = 6, 46 Hz, 1H), 7.66 (s, 1H), 7.44 (s, 1H), 7.30-7.41 (m, 3H), 6.59 (s, 1H), 5.61- 5.87 (m, 2H), 4.89 (d, J = 5.58 Hz, 2H), 4.58 (q, J = 7.35 Hz, 2H), 4.14 (br s, 2H ), 3.89 (t, J = 6.34 Hz, 2H), 3.84 (s, 3H), 3.25 (br s, 4H), 2.27 (t, J = 6.72 Hz , 2H), 2.21 (br s, 4H), 2.16 (s, 9H), 1.23-1.35 (m, 3H); LCMS (LCMS Method K): retention time = 0.78 min, [M + H] + = 818.4.

Stage 3:

tert-Butyl (E) -4- (3- (3-amino-5-carbamoyl-2 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido ) -7-methoxy-1 H-benzo ^] imidazol-1-yl) but-2-en-1-yl) amino) phenoxy) propyl) piperazine-1 carboxylate

The sodium salt of hydrosulfurous acid (371 mg, 1.81 mmol) was dissolved in H ₂ O (2 ml) at room temperature and then a solution of tert-butyl (E) -4- (3- (5-carbamoyl-2 - (( 4- (5-carbamoyl-2- (1ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy-1 H-benzo ^] imidazol-1-yl) but-2-ene-1 - yl) amino) -3 nitrophenoxy) propyl) piperazine-1-carboxylate (296 mg, 0.362 mmol) and ammonium hydroxide (0.48 6 ml, 3.62 mmol) in 5 ml MeOH. The reaction mixture was kept at room temperature for 2 h, then the reaction mixture was filtered and the filtrate was partially concentrated to remove MeOH. The resulting yellow aqueous mixture was then extracted with EtOAc 3 times, the organic extracts were combined and concentrated to give the title compound as a yellow solid (114 mg, 40% yield). 1H NMR (400 MHz, methanol ^ ₄ ) δ ppm 7.60 (d, J = 1.27 Hz, 1H), 7.31 (d, J = 1.27 Hz, 1H), 6.81 (d, J = 1.77 Hz, 1H), 6.67 (s, 1H), 6.59 (d, J = 1.77 Hz, 1H), 5.74-5.84 (m, 1H), 5, 53-5.65 (m, 1H), 4.12 (q, J = 7.18 Hz, 2 H), 3.84-3.91 (m, 3H), 3.61-3.71 (m , 4H), 3.38 (br s, 4H), 2.31-2.36 (m, 6H), 2.26 (s, 3H), 2.03 (s, 2H), 1.68- 1.78 (m, 2H), 1.47 (s, 9H), 1.42 (t, J = 7.10 Hz, 3H). LCMS (LCMS Method K): retention time = 0.65 min, [M + H] + = 788.5.

Stage 4:

tert-Butyl (E) -4- (3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- methoxy-1 H-benzo (d-imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) 1H-benzo ^] imidazole -7-yl) oxy) propyl) piperazine-1-carboxylate

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Tert-butyl (E) -4- (3- (3-amino-5-carbamoyl-2 - ((4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido ) -7-methoxy-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) amino) phenoxy) propyl) piperazine-1carboxylate (114 mg, 0.145 mmol) was dissolved in DMF (10 μl) at 0 ° C and then 1-ethyl-3methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.362 ml, 0.145 mmol) was added. The reaction mixture was kept at 0 ° C for 15 min, then TEA (0.050 mL, 0.362 mmol) and EDC (33.3 mg, 0.174 mmol) were added to the reaction mixture. The reaction mixture was kept at room temperature for 16 hours. The reaction mixture was then added to a stirred saturated NaHCO ₃ solution (aq). The resulting white precipitate was collected by filtration to give the title compound (103 mg, 75% yield). LCMS (LCMS Method K): retention time = 0.82 min, [M + H] + = 950.5.

Example 37 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (piperazin1-yl) propoxy) - 1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo ^ ] imidazole-5-carboxamide

Stage 5:

tert-Butyl (E) -4- (3 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- methoxy-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1H-benzo ^] imidazole- 7-yl) oxy) propyl) piperazine-1-carboxylate (103 mg, 0.109 mmol) was dissolved in MeOH (2 ml) and DCM (2 ml), then HCl (4N solution in 1,4-dioxane) (0.271 ml , 1.085 mmol). The reaction mixture was kept at room temperature for 16 h. Then DMSO (2 ml) was added to the reaction mixture and this mixture was filtered and the filtrate was concentrated and purified by HPLC (XSELECT CSH C18 column, 150 mm x 30 mm, package inner diameter 5 μm, 30 % -85% 10 mM ammonium bicarbonate in water with acetonitrile). The pure HPLC fractions were pooled and partially concentrated to give the title compound as a white precipitate (25 mg, 27% yield). 1H NMR (400 MHz, methanol-f) δ ppm 7.57 (d, J = 16.48 Hz, 2H), 7.14-7.30 (m, 2H), 6.50-6.70 (m , 2H), 5.81 (d, J = 3.04 Hz, 2H), 4.99 (br s, 4H), 4.50-4.69 (m, 4H), 3.86 (t, J = 5.70 Hz, 2H), 3.69 (s, 3H), 2.81 (t, J = 4.69 Hz, 4H), 2.32-2.36 (m, 6H), 2, 20 (d, J = 12.93 Hz, 6H), 1.70 (br s, 2H), 1.25-1.45 (m, 6H), LC-MS (LC-MS Method K): time retention = 0.67 min, [M + H] + = 849.8.

Example 38. (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) -1 H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7-ethoxy-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole -5-carboxamide

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Example 38 can be prepared according to method 20 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: (E) -1- (4 - ((2-amino-4carbamoyl-6- (3-morpholinopropoxy) phenyl) amino) but-2-en-1-yl) 7-ethoxy-2- ( 1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo (^] imidazole-5-carboxamide (46 mg, 0.065 mmol) was dissolved in DMF (655 μl) at 0 ° C, and then added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (196 μl, 0.079 mmol). The reaction solution was kept at 0 ° C. for 15 minutes, then EDC (15.06 mg, 0.079 mmol) and TEA (22.81 μl, 0.164 mmol) were added and the reaction solution was kept at room temperature. After 16 h, the reaction mixture was concentrated and the yellow residue was purified by HPLC (XSELECT CSH C18 column, 150 mm x 30 mm, 5 µm internal packing diameter, 15% -55% 10 mM ammonium bicarbonate in water with acetonitrile). The desired fractions were combined and concentrated to give the title compound as a white solid (19.2 mg, 34% yield). ¹ H NMR (400 MHz, methanol ^) δ ppm 7.62 (d, J = 1.27 Hz, 1H), 7.58 (d, J = 1.27 Hz, 1H), 7.24 (d, J = 1.27 Hz, 1H), 7.20 (d, J = 1.27 Hz, 1H), 6.64 (s, 1H), 6.62 (s, 1H), 5.78 (d, J = 3.30 Hz, 2H), 5.01 (d, J = 2.79 Hz, 4H), 4.63 (q, J = 7.10 Hz, 4H), 3.86-4.08 ( m, 6H), 3.69-3.81 (m, 2H), 3.37 (br s, 2H) 3.16-3.23 (m, 2H), 2.97 3.13 (m, 2H), 2.23 (s, 6H), 1.96-2.04 (m, 2H), 1.39 (t, J = 7.10 Hz, 6h), 1.15 (t, J = 6 , 97 Hz, 3H). LCMS (LCMS Method K): retention time = 0.76 min, [M + H] + = 864.5.

Example 3 9. (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1HbenzoI imidazol-1-yl) but-2-en-1-yl) -7- (3 - (dimethylamino) propoxy) -2- (1-ethyl-3-methyl-1 H-pyrazole-5carboxamido) -1H-benzo ^] imidazole-5- carboxamide dihydrochloride

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole -1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7 methoxy-1H-benzo ^] imidazole-5-carboxamide hydrochloride (150 mg, 0.184 mmol) in DMF (2 ml) was added TEA (0.20 ml, 1.435 mmol). The solution was cooled to 0 ° C. At this temperature, methanesulfonyl chloride (42.0 mg, 0.367 mmol) was added. The reaction mixture was stirred at this temperature for 1 hour, then another 1 equivalent of methanesulfonyl chloride (21.0 mg, 0.183 mmol) was added and the reaction was continued at 0 ° C for 1 hour. Then K ₂ CO ₃ (127 mg, 0.918 mmol) followed by the addition of 1 ml of dimethylamine (2 M solution in THF, 2.0 mmol). The reaction mixture was stirred at 80 ° C in a sealed tube for 2 h, then allowed to cool to room temperature and the crude material was purified by HPLC with mass separation. HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = 10 mM ammonium bicarbonate in H2O, adjusted with ammonia to pH 10, B = MeCN B%: 15-55. Collected the desired MW peaks. The solvent was removed and the residue was dissolved in 1 ml of MeOH. A 4N solution of HCl in dioxane (1 ml) was added. The solution was stirred at room temperature for 10 minutes. The solvent was removed and the solid was washed with ethyl ether (5 ml) to give the title compound (76 mg, 0.082 mmol, 44.7% yield). 1H NMR (DMSO-06, 600 MHz): δ ppm 12.89 (br s, 2H), 10.18-10.41 (m, 1H), 7.96-8.04 (m, 2H), 7.66 (d, J = 10.0 Hz, 2H), 7.35-7.41 (m, 2H), 7.28-7.35 (m, 2H), 6.53 (d, J = 2.8 Hz, 2H), 5.82 (dt, J = 15.5, 5.3 Hz, 1H), 5.71 (dt, J = 15.4, 5.6 Hz, 1H), 4, 85-4.98 (m, 4H), 4.52 (quint, J = 6.5 Hz, 4H), 3.96-4.04 (m, 2H), 3.70 (s, 3H), 3 , 00-3.09 (m, 2H), 2.66 (d, J = 4.8 Hz, 6H), 2.11 (d, J = 4.4 Hz, 6H), 1.85-2, 03 (m, 2H), 1.20-1.32 (m, 6H); LCMS Method K: retention time = 0.67 min, [M + H] + = 808.5.

Example 40 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (methylamino) propoxy) -1 H -benzo ^] imidazol-1-yl) but-2-en-1 -yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) 7-methoxy-1H-benzo ^] imidazole 5-carboxamide dihydrochloride

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To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1 H-benzo [d ] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7 methoxy-1H-benzo ^] imidazole-5 α-carboxamide hydrochloride (100 mg, 0.122 mmol) was added TEA (0.102 ml, 0.734 mmol). The solution was cooled to 0 ° C. Methanesulfonyl chloride (28.0 mg, 0.245 mmol) was added. The reaction mixture was stirred at 0 ° C for 1 h, then another 1 eq of MsCl was added and the reaction was continued at 0 ° C for 2 h. Then K ₂ CO ₃ (85 mg, 0.612 mmol) was added to the reaction mixture, followed by adding 1 ml of methanamine (2M solution in THF, 2.0 mmol). The reaction mixture was stirred at 80 ° C in a sealed tube for 2 h, then the reaction mixture was allowed to cool to room temperature and filtered. The crude filtrate was purified by mass separation HPLC. HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = 10 mM ammonium bicarbonate in H ₂ O, adjusted with ammonia to pH 10, B = MeCN B%: 15-55. The desired MW peaks were collected and the solvent was removed. The substance was dissolved in 2 ml of MeOH and 1 ml of a 4N solution of HCl in dioxane was added. The mixture was stirred at room temperature for 15 min, then concentrated to give the title compound (33 mg, 0.037 mmol, 30.2% yield). ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.90 (br s, 1H), 8.79 (br s, 2H), 8.03 (br s, 2H), 7.66 ( d, J = 7.10 Hz, 2H), 7.26-7.48 (m, 4H), 6.52 (s, 2H), 5.68-5.90 (m, 2H), 4.92 (dd, J = 17.24, 4.06 Hz, 4H), 4.52 (q, J = 6.76 Hz, 4H), 4.09 (t, J = 5.58 Hz, 2H) 3, 73 (s, 3H), 2.90 (d, J = 5.58 Hz, 2H), 2.46 (t, J = 5.32 Hz, 3H) s, 6H), 1.88-2.01 (m, 2H) 1.27 (t, J = 6.97 Hz, 6H); LCMS Method K: retention time = 0.66 min, [M + H] + = 794.4.

Example 41. (E) -7- (3-Aminopropoxy) -1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7-methoxy- 1 H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxamide ditrifluoroacetate

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1 H-benzo [d ] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7 methoxy-1H-benzo ^] imidazole-5 β-carboxamide dihydrochloride (100 mg, 0.117 mmol) was added TEA (0.049 ml, 0.351 mmol). The solution was cooled to 0 ° C. Methanesulfonyl chloride (0.014 ml, 0.176 mmol) was added and the reaction mixture was left to warm to room temperature and stirred at room temperature for 1 h. Then another 0.5 eq of MsCl was added and stirred at room temperature for 2 h, then another 0 was added. 5 equiv of MsCl and the reaction mixture was stirred for another 1 h. Then K ₂ CO ₃ (81 mg, 0.586 mmol) was added to the reaction mixture, followed by the addition of 7 M ammonia in MeOH (0.177 ml, 1.171 mmol). The reaction mixture was stirred at 50 ° C in a sealed tube overnight. The reaction mixture was then filtered and the crude filtrate was purified by mass separation HPLC to give the title compound (15.6 mg, 13.2%). HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = H ₂ O (0.1% TPA), B = MeCN (0.1% TPA) B%: 15-55. 1H NMR (400 MHz, DMSO-a ₆ ) δ ppm 12.86 (brs, 1H), 7.99 (brs, 2H), 7.76 (brs, 2H), 7.65 ( dd, J = 6.21, 0.89 Hz, 2H), 7.40 (brs, 2H), 7.28-7.36 (m, 2H), 6.51 (d, J = 8, 87 Hz, 2H), 5.77-5.88 (m, 4H), 4.91 (dd, J = 9.50, 4.44 Hz, 4H), 4.43-4.59 (m, 4H ), 4.10 (t, J = 5, 96 Hz, 2H), 3.72 (s, 3H) 2.832.97 (m, 2H), 2.11 (d, J = 5.07 Hz, 6H) , 1.90 (quint, J = 6.40 Hz, 2H), 1.2 6 (td, J = 7.10, 4.82 Hz, 6H); LCMS Method K: Retention Time = 0.65 min, [M + H] ⁺ = 780.5.

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Example 42. (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- (3-hydroxypyrrolidin-1-yl ) propoxy) -1 H-benzo ^] imidazol-1-yl) but-2-en-1 -yl) -2 - (1-ethyl-3-methyl-1 H-pyrazole 5-carboxamido) -7-methoxy- 1H-benzo ^] imidazole-5-carboxamide

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzoIimidazole-1- yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1H-benzo ^] imidazole-5-carboxamide dihydrochloride (100 mg, 0.117 mmol) in DMF (2 ml) was added TEA (71.1 mg, 0.703 mmol). The solution was cooled to 0 ° C. At this temperature, methanesulfonyl chloride (26.8 mg, 0.234 mmol) was added. The reaction mixture was stirred at this temperature for 45 minutes. Then another 1 eq of MsCl (13.5 mg) was added, stirring was continued at 0 ° C for 2 h. Then K ₂ CO ₃ (97 mg, 0.703 mmol) was added to the reaction mixture, followed by the addition of pyrrolidin-3-ol (102 mg, 1.17 mmol). The reaction mixture was stirred at 80 ° C in a sealed tube for 1 h, then the reaction mixture was allowed to cool to room temperature and the reaction mixture was filtered. The crude filtrate was purified by mass separation HPLC to give the title compound (56.6 mg, 0.063 mmol, 54.0% yield). HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = 10 mM ammonium bicarbonate in H ₂ O, adjusted with ammonia to pH 10, B = MeCN B%: 15-55. ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.84 (br s, 2H) 7.92-8.06 (m, 2H) 7.617.69 (m, 2H) 7.23-7.44 (m, 4H), 6.46-6.61 (m, 2H), 5.72-5.96 (m, 2H), 4.91 (dd, J = 13.05, 4.18 Hz, 4H ), 4.67 (br s, 1H), 4.52 (q, J = 7.18 Hz, 4 H), 4.12 (br s, 1H), 3.97 (t, J = 5 , 96 Hz, 2H), 3.35 (br s, 2H), 2.54-2.65 (m, 1H), 2.33-2.47 (m, 4H), 2.26 (br. s, 2H), 2.12 (d, J = 8.36 Hz, 6H) 1.81-1.96 (m, 1H) 1.60-1.76 (m, 2H) 1.46 (dd, J = 8.24, 4.69 Hz, 1H), 1.19-1.37 (m, 6H); LCMS Method K: retention time = 0.72 min, [M + H] + = 850.9.

Example 43. (E) -1 - ((E) -4 - ((E) -5-Carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -7- (3 (4- (2-hydroxyethyl) piperazin-1-yl) propoxy) -2,3-dihydro-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2 (( 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imine) -7-methoxy-2,3-dihydro-1H-benzo ^] imidazole-5carboxamide

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzoIimidazole-1- yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1H-benzo ^] imidazole-5-carboxamide hydrochloride (80 mg, 0.098 mmol) in DMF (2 ml) was added TEA (0.136 ml, 0.979 mmol) at room temperature. The solution was stirred at room temperature for 15 minutes, then methanesulfonic anhydride (51.1 mg, 0.294 mmol) was added at this temperature. The reaction mixture was stirred at this temperature for 45 minutes. Then, K ₂ CO ₃ (200 mg, 1.447 mmol) was added to the reaction mixture, followed by the addition of 2- (piperazine-1yl) ethan-1-ol (127 mg, 0.979 mmol). The reaction mixture was stirred at 80 ° C in a sealed tube for 1 h and at 50 ° C overnight, then the reaction mixture was allowed to cool to room temperature and filtered. HPLC crude filtrate with mass separation. Collected the desired MW peaks. Remove the solvent to give the title compound (45.1 mg, 0.051 mmol, 51.6% yield). HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = 10 mM ammonium bicarbonate in H ₂ O, adjusted with ammonia to pH 10, B = MeCN B%: 15-55. 1H NMR (400 MHz, DMSO-de) δ ppm 12.85 (brs, 2H), 7.98 (brs, 2H), 7.61-7.71 (m, 2H), 7.21 -7.45 (m, 4H) 6.47-4.65 (m, 4H), 4.34 (br s, 1H), 3.91 (t, J = 5, 96 Hz, 2H), 3 , 70 (s, 3H), 3.44 (q, J = 6.08 Hz, 2H), 2.18-2.93 (m, 10H), 2.14 (s,

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3H) 1.57-1.70 (m, 2H), 1.29 (q, J = 7.10 Hz, 6H); LC-MS Method K: retention time = 0.66 min,

[M + H] + = 893.4.

Example 44. (E) -1 - ((E) -4 - ((E) -5-Carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -7- (3 (3- (hydroxymethyl) morpholino) propoxy) -2,3-dihydro-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2 - ((1ethyl-3-methyl -1H-pyrazole-5-carbonyl) imino) -7-methoxy-2,3-dihydro-1H-benzo ^] imidazole-5-carboxamide ditrifluoroacetate

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole -1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1H-benzo ^] imidazole-5-carboxamide hydrochloride ( 100 mg, 0.122 mmol) in DMF (2 ml) was added TEA (0.20 ml, 1.435 mmol). The solution was cooled to 0 ° C and methanesulfonyl chloride (28.0 mg, 0.245 mmol) was added at this temperature. The reaction mixture was stirred at this temperature for 1 h. Then another 1 eq of MsCl was added, stirring was continued at 0 ° С for another 3 h. Then K ₂ CO ₃ (85 mg, 0.612 mmol) was added to the reaction mixture, followed by the addition of morpholine-3ylmethanol (86 mg, 0.734 mmol). The reaction mixture was stirred at 50 ° C in a sealed tube overnight, then the reaction mixture was allowed to cool to room temperature and filtered. The crude HPLC filtrate was mass-separated to give the title compound (13.1 mg, 9.66%). HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = H ₂ O (0.1% TPA), B = MeCN (0.1% TPA) B%: 15-55. 1H NMR (400 MHz, DMSO-06) δ ppm 12.49-13.47 (m, 1H), 9.64 (br s, 1H), 7.99 (d, J = 9.89 Hz, 2H ), 7.67 (d, J = 7.60 Hz, 2H), 7.40 (br s, 2H), 7.31 (d, J = 8.11 Hz, 2H), 6.54 (d , J = 1.77 Hz, 2H), 5.70-5.85 (m, 2H) 5.01 (m, 6H), 4.54 (dd, J = 6.72, 3.68 Hz, 4H ) 3.84-4.13 (m, 5H) 3.733.81 (m, 1H) 3.54-3.66 (m, 2H) 3.43 (m, (m, 5H), 2.12; LC -MS method K: retention time = 0.72 min, [M + H] ⁺ = 8 8 0.5.

Example 45 (E) -1 - ((E) -4 - ((E) -5-Carbamoyl-2 - ((1-ethyl-3-methyl-1 H-pyrazole-5-carbonyl) imino) -7methoxy -2,3-dihydro-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7- (3- (ethyl (2-methoxyethyl) amino) propoxy) -2 - (( 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imine) -2,3-dihydro-1H-benzo ^] imidazole-5-carboxamide

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole -1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1H-benzo ^] imidazole-5-carboxamide hydrochloride ( 100 mg, 0.122 mmol) in DMF (2 ml) was added TEA (0.171 ml, 1.224 mmol) at room temperature. At this temperature, methanesulfonic anhydride (42.6 mg, 0.245 mmol) was added. The reaction mixture was stirred at this temperature for 1 hour. Then another 1 equivalent of methanesulfonic anhydride was added, stirring was continued at room temperature for 30 minutes. Then K ₂ CO ₃ (169 mg, 1.224 mmol) was added to the reaction mixture, followed by the addition of N-ethyl-2-methoxyethan-1-amine (126 mg, 1.224 mmol). The reaction mixture was stirred at 80 ° C in a sealed tube, then the reaction mixture was cooled to room temperature and filtered. The crude filtrate was purified by mass separation HPLC (2 injections). Collect the desired MW peaks and remove the solvent to give the title compound (21.1 mg, 0.024 mmol, 19.52% yield). HPLC analysis was performed on a column

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XSELECT SCH C18. Eluents: A = 10 mM ammonium bicarbonate in H2O, adjusted with ammonia to pH 10, B = MeCN B%: 15-55. 1H NMR (400 MHz, DMSO-06) δ ppm 12.81 (brs, 2H), 7.98 (brs, 2H), 7.59-7.71 (m, 2H), 7.24 -7.42 (m, 4H) 6.46 (m, 2H), 5.75-5.94 (m, 2H), 4.92 (dd, J = 16.73, 4.06 Hz, 4H) , 4.47-4.42 (m, 4H), 3.96 (t, J = 5.83 Hz, 2 H), 3.72 (s, 3H) 3.24 (t, J = 6.08 Hz, 2H) 3.11 (s, 3H), 2.41 (t, J = 5.96 Hz, 4H), 2.34 (q, J = 7.10 Hz, 2 H), 2.12 ( d, J = 10.39 Hz, 6H), 1.55-1.66 (m, 2H), 1.29 (q, J = 7.10 Hz, 6H) 0.82 (t, J = 6, 97 Hz, 3H); LCMS Method K: retention time = 0.69 min, [M + H] + = 866.4.

Example 46. (E) -1 - ((E) -4 - ((E) -5-Carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl) imino) -7- (3 (4- (2-methoxyethyl) piperazin-1-yl) propoxy) -2,3-dihydro-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2 (( 1-ethyl-3-methyl-1 H-pyrazole-5-carbonyl) imino) -7-methoxy-2,3-dihydro-1H-benzo ^] imidazole-5-carboxamide tritrifluoroacetate

To a solution of (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1H-benzo ^] imidazole -1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7methoxy-1H-benzo ^] imidazole-5-carboxamide hydrochloride ( 100 mg, 0.122 mmol) in DMF (2 ml) was added TEA (0.171 ml, 1.224 mmol). The solution was cooled to 0 ° C. At this temperature, methanesulfonic anhydride (42.6 mg, 0.245 mmol) was added. The reaction mixture was stirred at this temperature for 1 h. Another equiv of MsCl was added, stirring continued at 0 ° C for 2 h. Then K ₂ CO ₃ (85 mg, 0.612 mmol) was added followed by the addition of 1- (2-methoxyethyl ) piperazine (176 mg, 1.22-4 mmol) and the reaction mixture was stirred at 50 ° C in a sealed tube overnight. The reaction mixture was then cooled to room temperature and filtered. HPLC crude filtrate with mass separation. Collect the desired MW peaks and remove the solvent to give the title compound (39.6 mg, 0.032 mmol, 25.9% yield). HPLC analysis was performed on an XSELECT SCH C18 column. Eluents: A = H ₂ O (0.1% TPA), B = MeCN (0.1% TPA) B%: 15-55. ¹ H NMR (400 MHz, DMSO) δ ppm 12.90 (br s, 2H) 8.00 (d, J = 9.89 Hz, 2H) 7.66 (s, 2H), 7.40 (d , J = 4.06 Hz, 2 H), 7.22-7.35 (m, 2H), 6.53 (d, J = 4.06 Hz, 2H), 5.70-5.89 (m , 2H), 4.91 (dd, J = 9.63, 4.56 Hz, 4H), 4.53 (dd, J = 7.10, 3.55 Hz, 6H), 3.94-4, 04 (m, 3H), 3.70 (s, 3H), 3.60 (d, J = 4.06 Hz, 2H) 3.31 (s, 3H), 3.09-3.25, 5H) , 2.86 (br s, 4H), 2.12 (d, J = 5.07 Hz, 6H) 1.74-1.88 (m, 2H) 1.28 (td, J = 7.10 , 3.30 Hz, 6H); LCMS Method K: retention time = 0.66 min, [M + H] ⁺ = 907.4.

Example 47 8-Ethyl-23 - ((4-methoxybenzyl) (methyl) amino) -10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20 , 21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1е: 4 ', 3' -1] [1,3,6,15,17] pentaazacyclogeneicosin-3 -carboxamide

- 113 036435

Stage 1:

3-Fluoro - \ - (4-methoxybenzyl) - \ - methyl-2-11itroa11 or11 F

MO ₂ 'RMV

A solution of 1- (4-methoxyphenyl) - N-methylmethanamine (5.23 g, 34.6 mmol) in CHCl ₃ (20 ml) was added to a stirred solution of 1,3-difluoro-2-nitrobenzene (5.5 g, 34, 6 mmol) and TEA (5.78 ml, 41.5 mmol) in CHCl ₃ (250 ml) at room temperature under N2 atmosphere. The mixture was stirred at room temperature for 30 min and then heated to 50 ° C overnight. Saturated NaHCO3 solution was added, the organic layer was separated, the aqueous layer was extracted with DCM, and the combined extracts were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (0-12% EtOAc in hexanes) to afford the title compound (8.5 g, 29.3 mmol, 8.5% yield) as an orange oil. ¹ H NMR (400 MHz, chloroform-d) δ ppm 7.29-7.34 (m, 1H) 7.20 (d, J = 8.28 Hz, 2H), 6.86-6.93 (m , 3H), 6.78 (t, J = 8.78 Hz, 1H) 4.29 (s, 2H), 3.82 (s, 3H), 2.80 (s, 3H); LCMS (LCMS Method D): retention time = 1.28, [M + H] + = 120.5.

Stage 2:

tert-Butyl (4 - ((3 - ((4-methoxybenzyl) (methyl) amino) -2-nitrophenyl) amino) butyl) carbamate n

NHBoc Y ^ no ₂ 'РМВ

To a solution of 3-fluoro-N (4-methoxybenzyl) N-methyl-2-nitroaniline (8.5 g, 29.3 mmol) in N, N-dimethylfirmolide (DMF) (60 ml) was added tert-butyl (4-aminobutyl) carbamate (8.27 g, 43.9 mmol) at room temperature and the mixture was stirred for 10 minutes; Then K2CO3 (8.09 g, 58.6 mmol) was added and the mixture was stirred at 80 ° C overnight. The reaction mixture was cooled to rooms

- 114 036435 temperature, diluted with EtOAc (200 ml) and washed sequentially with water (300 ml), 5% LiCl solution and brine. The organic layer was dried over Na ₂ SO ₄ , filtered, concentrated and the resulting residue was purified by silica gel column chromatography (0-30% EtOAc in hexane) to give the title compound (10.68 g, 23.29 mmol, 80% yield) in the form of red oil. 1H NMR (400 MHz, chloroform-d) δ ppm 7.09-7.28 (m, 3H), 6.87 (d, J = 7.53 Hz, 2H), 6.40 (br s, 1H ), 6.29 (d, 7.03 Hz, 1H), 4.57 (br s, 1H), 4.32 (br s, 2H), 3.81 (s, 3H), 3.11 -3.30 (m, 4H), 2.77 (br s, H) 1.67-1.79 (m, 2H) 1.55-1.67 (m, 2H) 1.47 (s, 9H). LCMS (LCMS Method D): retention time = 1.37, [M + H] + = 459.2.

Stage 3:

tert-Butyl (4 - ((2-amino-3 - ((4-methoxybenzyl) (methyl) amino) phenyl) amino) butyl) carbamate

Zinc (4.66 g, 71.3 mmol) was added portionwise to a stirred solution of tert-butyl (4 - ((3 - ((4methoxybenzyl) (methyl) amino) -2-nitrophenyl) amino) butyl) carbamate (10.9 g, 23.77 mmol) in acetic acid (200 ml). The mixture was stirred for 3 h and another portion of zinc (4.66 g, 71.3 mmol) was added. The mixture was stirred for an additional 30 minutes. The solid was filtered off and the filtrate was concentrated in vacuo. The residue was dissolved in DCM (200 ml), washed with 15% K ₂ CO ₃ solution and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and then concentrated in vacuo to afford the title compound (9.95 g, 23.22 mmol, 98% yield) as a brown foam which was used in the next step without purification. ¹ H NMR (400 MHz, methanol ^ ₄ ) δ ppm 7.24 (d, J = 8.53 Hz, 2H), 6.85 (d, J = 8.28 Hz, 2H), 6.66-6 , 73 (m, 1H), 6.57-6.63, 1H), 6.47 (d, J = 7.78 Hz, 1H), 3.90 (s, 2H) 3.78 (s, 3H ), 3.13 (dt, J = 13.30, 6.65 Hz, 4H), 2.53 (s, 3H), 1.58-1.76 (m, 4H) 1.45 (s, 9H ). LCMS (LCMS Method D): retention time = 1.00, [M + H] + = 429.2.

Stage 4:

tert-Butyl (4- (2-amino-4 - ((4-methoxybenzyl) (methyl) amino) -1H-benzo ^] imidazol-1-yl) butyl) carbamate hydrobromide

tert-Butyl (4-amino-3 - ((4-methoxybenzyl) (methyl) amino) phenyl) amino) butyl) carbamate (9.95 g, 23.22 mmol) was dissolved in methanol (80 ml), cyanogen bromide ( 4.64 ml, 23.22 mmol). The mixture was stirred for 18 h at room temperature, concentrated in vacuo to about 1/4 of the original volume, and MeCN (50 ml) and toluene (50 ml) were added. The mixture was concentrated to dryness and dried in vacuo for 16 hours to afford the title compound (12.54 g, quantitative yield) as a dark brown solid. LCMS (LCMS Method D): retention time = 1.04, [M + H] + = 454.2.

Stage 5:

Ethyl 4- (5- (5 - ((1- (4 - ((tert-butoxycarbonyl) amino) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1Нbenzo ^] imidazol-2-yl) carbamoyl) -3 -methyl-1 N-pyrazol-1 -yl) pentyl) -1-ethyl-3-methyl-1 N-pyrazole-5carboxylate

A mixture of 1- (5- (5 - (ethoxycarbonyl) -1-ethyl-3-methyl-1 N-pyrazol-4-yl) pentyl) -3 -methyl-1 N-pyrazole-5 carboxylic acid (1 g, 2 , 66 mmol), tert-butyl (4- (2-amino-4 - ((4-methoxybenzyl) (methyl) amino) -1Нbenzo ^] imidazol-1-yl) butyl) carbamate hydrobromide (1.704 g, 3.19 mmol), TEA (1.111 ml, 7.97 mmol), EDC (0.622 g, 3.45 mmol) and HOBt (0.610 g, 3.98 mmol) in NMP (25 ml) were stirred overnight at room temperature under atmosphere nitrogen. The reaction mixture was diluted with EtOAc (100 ml), washed

- 115 036435 saturated NaHCO ₃ solution, 5% LiCl solution and brine. The organic layer was dried over Na2SO4, filtered, concentrated and the residue was purified by silica gel column chromatography (0-45% EtOAc in DCM) to afford the title compound (1.3 g, 1.601 mmol, 60.3% yield) as a pink solid substances. LCMS (LCMS Method E): retention time = 1.56, [M + H] + = 812.6.

Stage 6:

Ethyl 4- (5- (5 - ((1- (4-aminobutyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1H-benzo ^] imidazol-2-yl) carbamoyl) -3-methyl -1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylate dihydrochloride

N HCl solution (2.232 ml, 8.93 mmol) was added to a stirred solution of ethyl 4- (5- (5 - ((1- (4 ((tert-butoxycarbonyl) amino) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1H-benzo (d] imidazole-2yl) carbamoyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylate (1 , 45 g, 1.786 mmol) in methanol (15 mL). The mixture was stirred overnight at room temperature under nitrogen atmosphere. The reaction mixture was concentrated in vacuo to about 1/5 of the original volume and MeCN (10 ml) and toluene (10 ml) were added. The mixture was concentrated to dryness and dried in vacuo to afford the title compound (1.45 g, 1.792 mmol, 100% yield) as an orange-red solid which was used in the next step without purification. Presumably a quantitative yield. LCMS (LCMS Method E): retention time = 1.19, [M + H] ⁺ = 712.6.

Stage 7:

Ethyl 4- (5- (5 - ((1- (4 - ((4-carbamoyl-2-nitrophenyl) amino) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) 1H-benzo ^] imidazole -2-yl) carbamoyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5carboxylate

To a solution of ethyl 4- (5- (5 - ((1- (4-aminobutyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1Нbenzo ^] imidazol-2-yl) carbamoyl) -3-methyl -1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5carboxylate dihydrochloride (100 mg, 0.124 mmol) in DMSO (1 ml) was added TEA (0.086 ml, 0.618 mmol) followed by adding 4-phenyl-3-nitrobenzamide (22.76 mg, 0.132 mmol) and the mixture was stirred at 70 ° C overnight. The reaction mixture was diluted with water, extracted three times with EtOAc, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash silica gel chromatography (EtOH / EtOAc 0-5%) to afford the title compound (74 mg, 0.084 mmol, 68.3% yield) as a yellow solid. ¹ H NMR (400 MHz, methanol-d) δ ppm 8.69 (s, 1H) 7.91 (d, J = 9.03 Hz, 1H), 7.18-7.31 (m, 3H) 7 , 09 (d, J = 8.03 Hz, 1 H), 6.96 (d, J = 9.04 Hz, 1H), 6.82-6.92 (m, 3H), 6.66 (s , 1H), 4.65 (t, J = 6.53 Hz, 2H), 4.20-4.43 (m, 8H), 3.76 (s, 3H), 3.48 (t, J = 6.27 Hz, 2H), 2.88 (s, 3H), 2.59 (t, J = 7.40 Hz, 2H), 2.21 (s, 3H), 2.11 (s, (m , 2H) 1.221.55 (m, 2H), 1.22-1.35 (m, 9H) LCMS (LCMS Method E): retention time = 1.41, [M + H] ⁺ = 877.5.

Stage 8:

Ethyl 4- (5- (5 - ((1- (4-carbamoylphenyl) amino) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1H-benzo ^] imidazol-2-yl) carbamoyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylate

- 116 036435

Zinc (0.240 g, 3, 66 mmol) was added in portions to a stirred solution of ethyl 4- (5- (5 - ((1- (4 ((4-carbamoyl-2-nitrophenyl) amino) butyl) -4 - ((4 -methoxybenzyl) (methyl) amino) -1 H-benzo ^] imidazol-2yl) carbamoyl) -3-methyl-1H-pyrazol-1 -yl) pentyl) -1 -ethyl-3 -methyl-1H-pyrazole-5 β-carboxylate (1.07 g, 1.221 mmol) in acetic acid (10 ml). The mixture was stirred for 30 min and another portion of zinc (0.240 g, 3.66 mmol) was added. The mixture was stirred for an additional 30 min, the solid was filtered off and the filtrate was concentrated in vacuo. The residue was dissolved in DCM (50 ml) and washed with 15% K ₂ CO ₃ solution and then with brine. The organic layer was dried over Na ₂ SO ₄ , filtered and then concentrated in vacuo to give the title compound (1.16 g, 1.244 mmol) as a brown foam, which was used in the next step without purification. Presumably a quantitative yield. LCMS (LCMS Method E): retention time = 1.29, [M + H] + = 847.6.

Stage 9:

Ethyl 4- (5- (5 - ((1 - (4- (2-amino-5-carbamoyl-1 H-benzo ^] imidazol-1 -yl) butyl) -4 - ((4-methoxybenzyl) (methyl ) amino) -1 H-benzo (^] imidazol-2-yl) carbamoyl) -3 -methyl-1 H-pyrazol-1-yl) pentyl) -1-ethyl-3methyl-1 H-pyrazole-5-carboxylate

Ethyl 4- (5- (5 - ((1- (4 - ((2-amino-4-carbamoylphenyl) amino) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) 1 H-benzo ^] imidazol-2-yl) carbamoyl) -3-methyl-1 N-pyrazol-1 -yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5 carboxylate (1.03 g, 1.277 mmol) was dissolved in methanol (5 ml) and cyanogen bromide (0.243 ml, 1.217 mmol) was added. The mixture was stirred for 18 hours at room temperature. The reaction mixture was concentrated, the residue was dissolved in 10% MeOH solution in DCM (100 ml) and washed with 10% K ₂ CO ₃ solution and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography on silica gel (2M NH ₃ in MeOH / DCM 0-10%) to afford the title compound (870 mg, 0.999 mmol, 82% yield) as a light violet foam. 1H NMR (400 MHz, methanol X ₄ ) δ ppm 7.75 (s, 1H), 7.55 (d, J = 8.28 Hz, 1H), 7.18-7.99 (m, 3H), 7 , 15 (d, J = 8.28 Hz, 1H), 7.00 (d, J = 8.28 Hz, 1H), 6.81-6.93 (m, 3H), 6.65 (s, 1H), 4.62 (t, J = 6.65 Hz, 2H), 4.19-4.41 (m, 8H) 4.03-4.13 (m, 2 H), 3.75 (s , 3H), 3.37 (s, 1H), 2.88 (s, 3H) 2.58 (t, J = 7.40 Hz, 2H), 2.24 (s, 3H) 2.10 (s , 3H), 1.77-2.00 (m, 6H), 1.42-1.54 (m, 2H), 1.27 (t, J = 7.03 Hz, 9H). LCMS (LCMS Method E): retention time = 1.20, [M + H] + = 872.5.

Stage 10:

4- (5- (5 - ((1 - (4- (2-Amino-5-carbamoyl- 1H-benzo ^] imidazol-1 -yl) butyl) -4 - ((4-methoxybenzyl) (methyl) amino ) -1H-benzo (^] imidazol-2-yl) carbamoyl) -3 -methyl-1H-pyrazol-1 -yl) pentyl) -1 -ethyl-3-methyl-1Hpyrazole-5-carboxylic acid

- 117 036435

To a suspension of ethyl 4- (5- (5 - ((1- (4- (2-amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) butyl) -4 - ((4-methoxybenzyl) ( methyl) amino) -1H-benzo (| imidazol-2-yl) carbamoyl) -3 -methyl-1H-pyrazol-1 -yl) pentyl) -1 ethyl 3-methyl-1H-pyrazole-5-carboxylate ( 870 mg, 0.999 mmol) in methanol (8 ml) was added NaOH (4.99 ml, 4.99 mmol) and the resulting clear homogeneous solution was stirred overnight at room temperature. Methanol (8 ml) was added followed by dropwise addition of 1 M HCl solution (5 ml). The mixture was concentrated to remove most of the methanol and water was added. The particles were filtered, washed with water, air dried and then dried in vacuo to afford the title compound (782 mg, 0.928 mmol, 93% yield) as a light pink solid. ¹ H NMR (400 MHz, DMSO ^) δ ppm 7.83 (br s, 1H), 7.71 (s, 1H), 7.53 (d, J = 8.07 Hz, 1H), 7.017, 22 (m, 7H) 6.84 (d, J = 8.31 Hz, 2H), 6.67 (br s, 1H), 4.42 (br s, 2H), 4.34 (q, J = 7.09 Hz, 2H), 4.12 (br s, 2H), 3.97-4.06 (m, 2H), 2.85 (s, 3H), 2.52-2.56 (m, 2H), 2.15 (s, 3H), 2.04 (s, 3H), 1.81 (br s, 2H) 1.70 (d, J = 6.36 Hz, 4H) , 1.34-1.46 (m, 2H), 1.14-1.27 (m, 6H). LCMS (LCMS Method E): retention time = 1.02, [M + H] + = 843.5.

Example 47 8-Ethyl-23 - ((4-methoxybenzyl) (methyl) amino) -10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20 , 21,28, 29, 30, 31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-3-carboxamide

A solution of 4- (5- (5 - ((1- (4- (2-amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) butyl) -4 - ((4-methoxybenzyl) (methyl) amino) -1H-benzo (d) imidazol-2-yl) carbamoyl) -3 -methyl-1H-pyrazol-1 -yl) pentyl) -1 -ethyl-3 methyl-1H-pyrazole-5-carboxylic acid (200 mg, 0.377 mmol) in DMF (5 ml) was slowly added (8 h) with a syringe pump to a mixture of 1H-benzo [c1] [1,2,3] triazol-1-ol (64.1 mg, 0.475 mmol), DMAP (2.90 mg, 0.024 mmol) and EDC (68.2 mg, 0.356 mmol) in DMF (5 ml) at 60 ° C under nitrogen atmosphere. The reaction mixture was stirred for another 24 h, concentrated in vacuo and the residue was dissolved in 10% MeOH solution in DCM. Saturated aqueous NaHCO ₃ solution was added and the mixture was extracted with 10% MeOH solution in DCM. The combined extracts were washed with brine, dried over Na ₂ SO ₄ , filtered, concentrated and the residue was purified by flash chromatography on silica gel (2M NH ₃ in MeOH / DCM 0-10%) to give the title compound (40 mg, 0.048 mmol, 20.44% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.88 (s, 2H) 8.02 (d, J = 1.47 Hz, 2H), 7.82 (d, J = 8.56 Hz, 1H) 7.49-7.65 (m, 1H), 7.38 (br s, 1H), 7.20 (br s, 3H), 6.85 (br s, 3H), 6 , 50-6.64 (m, 1H), 4.54-5.18 (m, 3H) 4.48 (d, J = 7.09 Hz, 2H), 4.20 (brs, 5H) , 3.71 (s, 3H), 2.72-2.91 (m, 5H), 2.18 (br s, 3H), 1.77-1.97 (m, 6H), 1.49 (br s, 2H), 1.30 (t, J = 7.09 Hz, 5H). LCMS (LCMS Method D): retention time = 1.18, [M + H] + = 825.8.

Examples 48-50.

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Example 48 8-Ethyl-10,18-dimethyl-23- (methylamino) -7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,

29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogenaicosin-3-carboxamide

Pd-C (0.258 g, 0.242 mmol) was added to a solution of 8-ethyl-23 - ((4-methoxybenzyl) (methyl) amino) -10.18dimethyl-7.20-dioxo-6,7,8,11,12 , 13,14,15,20,21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5 , 1st: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogenaicosine-3-carboxamide (2.0 g, 2.424 mmol) in MeOH (20 ml) and DCM (20 ml) ... The flask was purged with nitrogen, then with hydrogen, and the mixture was stirred under a hydrogen atmosphere. After 8 h, Pd / C was filtered off and the filtrate was concentrated in vacuo to give a white solid, which was washed with MeOH to give the title compound (1.25 g, 1.773 mmol, 73.2% yield) as a white solid. 1H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.86 (br s, 1H) 12.53 (br s, 1H), 8.02 (br s, 1H), 7.99 (br. s, H), 7.81 (d, J = 8.07 Hz, 1H), 7.58 (d, J = 8.07 Hz, 1H), 7.34 (brs, 1H), 7, 13 (t, J = 7.70 Hz, 1H), 6.83 d, J = 7.82 Hz, 1H), 6.72 (d, J = 4.16 Hz, 1H), 6.55 (s , 1H), 6.41 (d, J = 7.83 Hz, 1H), 4.75 (br s, 2H), 4.48 (d, J = 6.85 Hz, 2H), 4.22 (br s, 4H), 2.82 (d, J = 3.91 Hz, 5H) 2.16 (br s, 3H), 2.09 (br s, 3H), 1.90 (br s, 4H), 1.81 (br s, 2H), 1.49 (br s, 2H), 1.38 (br s, 2H) 1.30 (t, J = 6.85 Hz , 3H); LCMS (LCMS Method D): retention time = 1.01, [M + H] + = 705.5.

Example 49.2 - ((3-Carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29 , 30, 31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [ 1,3,6,15,17] pentaazacycloneicosin-23-yl) (methyl) amino) -2-oxoethyl acetate

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2-Chloro-2-oxoethyl acetate (0.069 ml, 0.638 mmol) was added to the suspension of 8-ethyl-10,18-dimethyl-23 (methylamino) -7,20-dioxo-6,7,8,11,12,13, 14,15,20,21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2а] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacycloneicosine 3-carboxamide (300 mg, 0.426 mmol) in pyridine (5 ml) at room temperature. The mixture was stirred overnight, then concentrated and the residue was dissolved in 10% MeOH in DCM (100 ml) and washed with water and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography on silica gel (2M NH ₃ in MeOH / DCM 0-10%) to afford the title compound (198 mg, 0.246 mmol, 57.8% yield) as a light pink solid. 1H NMR (400 MHz, methanol ^ ₄ ) δ ppm 7.97 (s, 1H) 7.86 (d, J = 8.28 Hz, 1H) 7.59 (d, J = 6.78 Hz, 1H) 7.38-7.53 (m, 2 H), 7.34 (d, J = 7.78 Hz, 1H), 6.60-6.75 (m, 1H), 4.76 (br s , 2H), 4.48-4.60 (m, 3H), 4.31 (bs, 5H), 3.54 (bs, 1H), 3.36 (s, 2H), 2, 89 (brs, 2H), 2.26 (brs, 3H), 2.18 (s, 3H), 2.04 (brs, 6H), 1.96 (s, 1H), 1.88 (d, J = 6.53 Hz, 2H) 1.60 (brs, 2H), 1.33-1.49 (m, 5H); LCMS (LCMS Method D): retention time = 0.94, [M + H] + = 805.6.

Example 50 8-Ethyl-23- (2-hydroxy-N-methylacetamido) -10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21, 28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1е: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogenaicosin-3-carboxamide

To a solution of 2 - ((3-carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29, 30, 31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1 , 3,6,15,17] pentaazacyclogenaicosin-23-yl) (methyl) amino) -2-oxoethyl acetate (132 mg, 0.164 mmol) in MeOH (4 ml), lithium hydroxide (1.640 ml, 1.640 mmol) was added and the mixture was stirred at room temperature. After 3 h, 1.64 ml of 1 M HCl solution was added to the suspension to obtain a clear solution. Most of the MeOH was removed in vacuo, water was added and the solid was isolated by filtration, washed with water, air dried and then dried in vacuo to give the title compound (119 mg, 0.156 mmol, 95% yield) as a white solid. 1H NMR (400 MHz,

DMSO ^ 6) δ ppm 12.87 (s, 2H), 7.93-8.06 (m, 2H), 7.83 (d, J = 7.58 Hz, 1H), 7.50-7, 71 (m, 2H), 7.35 (br s, 2H), 7.25 (d, J = 7.09 Hz, 1H), 6.56 (br s, 1H), 4.54-4 , 88 (m, 3H), 4.49 (q, J = 7.01 Hz, 2 H), 4.24 (bs, 5H), 3.49-3.88 (m, 2H), 3 , 22 (br s, 2H), 2.82 (br, 2H), 2.16 (br) 2.10 (s, 3H) 1.69-2.02 (m, 6H) 1.50 (br s, 2H) 1.31 (t, J = 7.09 Hz, 5H); LCMS Method D: retention time = 0.88 min, [M + H] + = 763.6.

Example 51 (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5 -carboxamido) -1 Nbenzo ^] imidazol-1-yl) but-2 -en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (2- (tetrahydro2H-pyran-4-yl) ethoxy) -1 H-benzo ^ ] imidazole-5-carboxamide

Example 51 can be prepared according to method 14 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: a mixture of 4- (2-bromoethyl) tetrahydro-2H-pyran (12.54 mg, 0.065 mmol), (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3 -methyl-1H-pyrazole-5carboxamido) -1 H-benzo (^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -7-hydroxy-1H-benzo ^] imidazole-5-carboxamide (45 mg, 0.065 mmol) and potassium carbonate (22.44 mg, 0.162 mmol) were heated for 3 h at 85 ° C in DMSO (650 μl) and NMP (650 μl), then cooled.

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The residue was purified by acidic reversed phase chromatography (5-50% in 0.1% TFA in MeCN to 0.1% TFA in water, Phenomenex Eclipse C18 column, 50 mm x 30 mm, 20 min gradient). The pure fractions were partitioned between EtOAc and aqueous saturated sodium bicarbonate, the organic layer was separated, dried over sodium sulfate and evaporated in vacuo to give the title compound (8 mg, 15.3% yield) as a white solid. ¹ H NMR (DMSO-d ⁶ , 600 MHz): δ ppm 12.83 (brs, 2H), 7.97-8.00 (m, 1H), 7.93 (brs, 2H), 7.69 (dd, J = 8.4, 1.5 Hz, 1H), 7.63 (s, 1H), 7.41 (d, J = 8.3 Hz, 1H), 7.33 (br d, J = 11.4 Hz, 2H), 7.29 (s, 1H) 6.55 (s, 1H), 6.52 (s, 1H), 5.96-6.02 (m, 1H ), 5.70-5.79 (m, 1H), 4.93 (brd, J = 5.0 Hz, 2H), 4.82 (brd, J = 5.3 Hz, 2H) , 4.49-4.58 (m, 4H), 3.96 (brt, J = 6.7 Hz, 2H), 3.75 (brd, J = 11.2, 2.9 Hz , 2H), 3.16-3.23 (m, 2H), 2.12 (d, J = 12.7 Hz, 6H), 1.50-1.53 (m, 1H), 1.45- 1.49 (m, 2H), 1.43 (brd, J = 11.9 Hz, 2H), 1.28 (m, 6H), 1.08 (brdd, J = 12.0, 3.6 Hz, 2H); LCMS (LCMS Method K): retention time = 0.90 min, [M + H] + = 805.5.

Example 52. (E) -4 - ((5-Carbamoyl-1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) 1 H-benzo ^ imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) - 1H-benzo [d] imidazol-7-yl) oxy ) butanoic acid he

Example 24 can be prepared according to a combination of method 14 and 16 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: methyl (E) -4 - ((5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H- benzoU] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) - 1H-benzo Schimidazol-7-yl) oxy ) butanoate (40 mg, 0.050 mmol) was dissolved in MeOH and THF (1 ml each) and sodium hydroxide (101 μl, 0.505 mmol, 5 N solution) was added, and the mixture was stirred at 25 ° С for 18 h. Then the reaction the mixture was distributed between EtOAc and 10% aqueous potassium hydrogen sulfate solution. The resulting sticky gel mixture was evaporated to near dryness, dissolved in 2 ml of MeOH with aqueous sodium hydroxide solution (5 N solution) for dissolution. The residue was purified by basic reverse phase chromatography (10-55% in 0.1% NH ₄ OH in water to MeCN, Phenomenex Gemini C18 110A column, 50 mm x 30 mm, 10 min gradient). Pure fractions were collected and the product was isolated by concentration in vacuo, then dried under high vacuum to give the title compound as a white solid. 1H NMR (400 MHz, methanol ^ ₄ ) δ ppm 8.45-8.76 (m, 1H), 7.85-8.12 (m, 1H), 7.49-7.78 (m, 2H) , 6.92-7.30 (m, 2 (M, 1H), 4.45-4.66 (m, 5H), 3.91-4.166 (m, 4H) H), 3.6 (q, J = 6.3 Hz, 4H), 2.31 (m, 2H), 2.18 (s, 6H), 1.29 (q, J = 6, 1 Hz, 4H), 1.13 (t, J = 6.1 Hz, 6H); LCMS Method K: retention time = 0.75 min, [M + H] + = 779.4.

Example 53.3 - (((2) -6-Carbamoyl-3 - ((E) -4 - ((2) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl ) imino) -2,3-dihydro- 1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1H-pyrazol5- carbonyl) imino) -2,3-dihydro-1H-benzoU] imidazol-4-yl) oxy) propyldihydrogen phosphate dihydrochloride

Stage 1:

Ditert-butyl (3 - (((2) -6-carbamoyl-3 - ((E) -4 - ((2) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole- 5carbonyl) imino) -2,3-dihydro-1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1H-pyrazole5-carbonyl) imino) -2,3-dihydro-1H-benzoI imidazol-4-yl) oxy) propyl) phosphate

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(E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoH | imidazole-1yl) but-2-yn-1- yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-hydroxypropoxy) -1HbenzoA | imidazole-5-carboxamide (100 mg, 0.133 mmol) was suspended in DMF (1 ml), 2H tetrazole in MeCN (1.480 ml, 0.666 mmol) was added and the mixture was concentrated on a rotary evaporator to remove the MeCN. The reaction mixture was cooled in an ice water bath then a solution of di-tert-butyldiisopropylphosphoramidite (148 mg, 0.533 mmol) in DMF (1 ml) was added. The mixture was stirred for 1 h, slowly warmed to room temperature and left overnight for 16 h, then cooled in an ice water bath, and an additional amount of 2 eq of 2H-tetrazole in MeCN and 2 eq of di-tert-butyldiisopropylphosphoramidate was added and the reaction mixture was stirred for 2 h. Then an additional 2 eq of 2H-tetrazole in MeCN and 2 eq of di-tert-butyldiisopropylphosphoramiditate were added and the reaction mixture was stirred for 2 h. Then the reaction mixture was cooled in a bath with ice water, H ₂ O ₂ (0.272 ml, 2 , 66 mmol, 30%) and stirring was continued for 30 min. The reaction mixture was poured into water (50 ml) containing a mixture of NaHCO ₃ and Na ₂ S2O ₃ (1: 1, 2M, 1 ml). The sticky paste was filtered off, washed with water, and dried on the filter for 2 days. The residue was dissolved in THF, combined with the extracts from the filtrate using a 3: 1 CHCl ₃ : EtOH mixture, the organic layers were dried over MgSO ₄ , concentrated, dried over silica gel (12 g column) and purified by silica gel chromatography using a 1-10% mixture MeOH in DCM (+ 1% NH ₄ OH) for 5 min, then 10% for 15 min to afford the title compound (23 mg, 0.024 mmol, 18.31% yield) as a white solid. The compound was used in the next step directly without purification. LCMS (LCMS Method I): retention time = 1.03 min, [M + H] ⁺ = 943.3.

Example 53.3 - (((2) -6-Carbamoyl-3 - ((E) -4 - ((Z) -5-carbamoyl-2 - ((1-ethyl-3-methyl-1H-pyrazole-5carbonyl ) imino) -2,3-dihydro-1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1H-pyrazole 5-carbonyl ) imino) -2,3-dihydro-1H-benzo-imidazol-4-yl) oxy) propyldihydrogen phosphate dihydrochloride

Di-tert-butyl (3 - ((^) - 6-carbamoyl-3 - ((E) -4 - ((Z) -5-carbamoyl-2 - ((1-ethyl-3 -methyl-1Н-pyrazole -5carbonyl) imino) -2,3-dihydro-1H-benzo (imidazol-1-yl) but-2-en-1-yl) -2 - ((1-ethyl-3-methyl-1H-pyrazole 5-carbonyl ) imino) -2,3-dihydro-1H-benzo-imidazol-4-yl) oxy) propyl) phosphate (22 mg, 0.023 mmol) was suspended in dioxane (1 ml), a 3M HCl solution in CPME (0.156 ml, 0, 4 67 mmol at room temperature). After 2 h, add diethyl ether (50 ml), filter the precipitate under nitrogen, wash with diethyl ether, dry in a vacuum oven at 40 ° C for 2 h to give the title compound (18 mg, 0.020 mmol, 85% yield) as a white solid. '11 NMR (700 MHz, DMSO-de) δ ppm 12.83 (br s, 2H), 7.92-8.05 (m, 4H), 7.73 (dd, J = 8.4, 1 , 4 Hz, 1H), 7.64-7.67 (m, 1H), 7.43 (d, J = 8.3 Hz, 2H), 7.30-7.39 (m, 4H), 6 , 55 (s, 1H), 6.49 (s, 1H), 6.02 (dt, J = 15.4, 5.5 Hz, 1H), 5.62-5.77 (m, 1H), 4.95 (br d, J = 4.8 Hz, 3H), 4.83 (br d, J = 5.4 Hz, 3H), 4.44-4.59 (m, 9H), 4 , 15 (br t, J = 6, 1 Hz, 5H), 3.94-3.99 (m, 4H), 2.11 (s, 4H), 2.09 (s, 3H), 1, 96 (quin, J = 6.0 Hz, 3H), 1.25 (quint, J = 7.3 Hz, 8H); LCMS (LCMS Method I): retention time = 0.64 min, [M + H] ⁺ = 831.2.

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Example 54 3-Carbamoyl-8-ethyl-10,18-dimethyl-7,2 0-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30, 31 tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3, 6,15,17] pentaazacyclogeneicosine-24-carboxylic acid

Stage 1:

Methyl 3-carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13, -14,15,20,21,28,29,30,31- tetradecahydrobenzo [4,5] imidazo [1,2а] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15 , 17] pentaazacyclogeneicosine-24-carboxylate

Example 54 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to a suspension of 24-cyano-8ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30 , 31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1, 3,6,15,17] pentaazacyclogenaicosine-3carboxamide (900 mg, 1.284 mmol) in MeOH (40 ml) was added boron trifluoride etherate (0.814 ml, 6.42 mmol). The reaction mixture was stirred at 80 ° C for 48 h, then concentrated in vacuo and poured into ice water. The precipitate was collected by filtration, washed with water and dried to give the title compound (600 mg, 0.818 mmol, 63.7% yield) as a gray solid. LCMS (LCMS Method A): retention time = 1.406 min, [M + H] + = 733.7.

Example 54 3-Carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31 tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6, 15,17] pentaazacyclogeneicosine-24-carboxylic acid

To a suspension of methyl 3-carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21, 28,29, 30,31 -tetradecahydrobenzo [4,5] imidazo [1,2а] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6, 15.17] pentaazacyclogeneicosine-24-carboxylate (420 mg, 0.572 mmol) in MeOH (15 ml) and water (15 ml), NaOH (22 9 mg, 5.72 mmol) was added. The reaction mixture was stirred at 25 ° C for 16 hours. The mixture was diluted with water (20 ml), acidified with 2N HCl solution to pH = 3 and the precipitate was collected by filtration to give the crude product. The crude product was purified by preparative HPLC (OBD Gemini Prep C18 column, 5 μm silica gel, diameter 21.2 mm, length 150 mm) using a mixture of decreasing polarity water (containing 0.1% TFA) and MeCN as eluants. Fractions containing the title compound were evaporated to dryness to afford the title compound (190 mg, 0.244 mmol, 46.1% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSOD6) δ ppm 12.89 (s, 1H), 8.13 (d, J = 9.5 Hz, 1H), 7.87 (d, J = 8.4 Hz, 1H ), 7.71-7.52, 3H), 7.45 (d, J = 4.7 Hz, 1H), 6.55 (s, 1H), 4.51 (q, J = 6.8 Hz , 2H), 4.36 (t, J = 7.0 Hz, 2H), 4.17, 2H), 4.10 (d, J = 6.8 Hz, 2H), 2.65 (t, J = 7.3 Hz, 2H), 2.10 (t, J = 9.2 Hz, 6H), 1.87-1.61 (m, 6H), 1.46 (s, 2H), 1.28 (t, J = 7.0 Hz, 3H), 1.19 (s, 2H); LCMS Method A: retention time = 1.2 95 min, [M + H] + = 720.2.

Example 55. Methyl 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo Schimidazol-1-yl) butyl) -2- (1-ethyl -3-methyl-1Н-pyrazole-5-carboxamido) -7-hydroxy-1Нbenzo H | imidazole-5-carboxylate di-trifluoroacetate ° x ^ NH

Example 55 can be prepared according to method 11 with modifications known to one of ordinary skill in the art. The last stage of preparation is presented: in a round-bottom flask with a capacity of 50 ml methyl 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoShimidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo-imidazole-5-carboxylate (35 mg, 0.048 mmol) was stirred in DCM. Boron tribromide (58.0 μl, 0.058 mmol) was then added portionwise at room temperature. The reaction mixture was stirred at room temperature overnight (about 18 h), after which the reaction was quenched with MeOH and the reaction mixture was concentrated in vacuo. The crude product, containing both the methoxy derivative and the phenolic derivative, was purified by reversed phase HPLC (Gilson 115 solution machine, Gilson 333 aqueous liquid pump, Gilson 334 organic liquid pump, Gilson UV / VIS-155 detector, Trilution v1.4 software) Luna column: acetonitrile, 0.1% TFA / water, gradient 0-20% eluent The desired fractions were combined and dried in vacuo to give the title compound (5 mg, 5.07 μmol, 10.47% yield). ¹ H NMR (400 MHz, DMSO-d ^ δ ppm 12.62-13.04 (m, 2H) 10.69 (s, 1H), 7.99 (s, 2H), 7.72-7.82 (m, 1H), 7.65 (s, 1H), 7.54 (d, J = 8.34 Hz, 1H), 7.34 (d, J = 2.27 Hz, 2H), 6.59 (s, 2H), 4.57 (dd, J = 6.82, 4.04 Hz, 5H), 4.44 (br s, 3H), 3.80-3.90 (m, 3H), 2.10 (d, J = 4.55 Hz, 6H), 1.89 (br s, 4H), 1.31 (t, J = 7.07 Hz, 6H), LC-MS (LC-MS method C): retention time ^, 87 min, [M + H] + = 710.6.

Example 56 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoSHimidazol-1-yl) but-2-ene -1-yl) -2- (1-ethyl-4-fluoro-3-methyl-1H-pyrazole-5-carboxamido) -7- (3-morpholinopropoxy) -1H-benzoSimidazole-5-carboxamide

Example 56 can be prepared according to method 20 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to a suspension of (E) -2-amino1- (4- (5) carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoSchimidazol-1-yl ) but-2-en-1yl) -7- (3-morpholinopropoxy) -1H-benzo-thimidazole-5-carboxamide (50 mg, 0.073 mmol) in DMF (2 ml), a solution of 1-ethyl-3-fluoro-4- methyl-1H-pyrazole-5-carboxylic acid (31.5 mg, 0.183 mmol), HOBt (16.8 mg, 0.110 mmol), HATU (69.5 mg, 0.183 mmol) and triethylamine (0.04 ml, 0 , 3 mmol) in DMF (2 ml) at room temperature. The mixture was stirred overnight, then heated to 50 ° C for 30 minutes. Water was added and the cloudy solution was cooled in a refrigerator until a precipitate settled. Tver

124 036435 the final material was collected by filtration and purified on silica gel (Isco silica gel 4 g column) eluting with 0-20% MeOH in DCM to give the title compound (4 mg, 4.77 μmol, 6.5% yield) as white solid. 1H NMR (400 MHz, DMSO-de) 5 ppm 12.81 (brs, 2H), 7.99-7.33 (m, 1H), 7.92-7.97 (m, 1H), 7 , 67-7.73 (m, 1H), 7.64 (s, 4H), 7.30-7.44 (m, 4H), 6.53 (br s, 1H), 5.93-6 , 04 (m, 1H), 5.68-5.82 (m, 1H), 4.90-4.97 (m, 2H), 4.76-4.84 (m, 2H), 4.45 -4.57 (m, 4H), 3.98 (br s, 2H), 3.44-3.49 (m, 4H), 2.21-2.29 (m, 2H), 2.17 -2.21 (m, 4H), 2.07-2.15 (m, 6H), 1.63-1.74 (m, 2H), 1.13-1.21 (m, 6H); LCMS (LCMS Method J): retention time = 0.65 min, [M + H] + = 838.3.

Example 57 1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoShimidazol-1-yl) butyl) -2- (1-ethyl -3 -methyl-1H-pyrazole-5-carboxamido) -7-hydroxy-1H-benzo Schimidazole-5 carboxamide trifluoroacetate

Example 57 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to 1- (4- (5-carbamoyl-2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo (^ imidazol-1-yl) butyl) -2- ( 1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) -7-methoxy-1H-benzoHimidazole-5-carboxamide trifluoroacetate (17 mg, 0.018 mmol) in DCM (2 ml) at -78 ° C was added 1 M BBr solution ₃ in DCM (50 μl, 0.050 mmol). After 1 hour, the reaction mixture was warmed to room temperature and LC-MS indicated the reaction was complete. The reaction mixture was cooled to 0 ° C and an additional 100 μl of 1 M solution of BBg ₃ in DCM was added. The reaction mixture was allowed to slowly warm to room temperature over 72 h, returned to 0 ° C, then another 100 μl of 1 M BBr ₃ in DCM was added. After another 24 h, MeOH (2 ml) was added and the reaction mixture was concentrated. The residue was purified by reverse phase HPLC eluting with 20-45% MeCN in H ₂ O (0.1% TFA) to give the title compound (8 mg, 0.01 mmol, 55% yield). 1H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.80 (br s, 1H), 10.48 (s, 1H), 7.98 (d, J = 1.27 Hz, 2H), 7.83 (br s, 1H), 7.75 (dd, J = 8.36, 1.52 Hz, 1H), 7.83 (br s, 1H), 7.75 (dd, J = 8.36 , 1.52 Hz, 1H), 7.53 (d, J = 8.36 Hz, 1H), 7.43 (d, J = 1.27 Hz, 1H), 7.22 (s, 1H), 7.34 (br s, 1H), 7.14 (d, J = 1.52 Hz, 1H), 7.10 (s, 1H), 6.97 (s, 1H), 6.58 (d , J = 9.63 Hz, 2H), 4.49-4.65 (m, 4H), 4.43 (t, J = 6.40 Hz, 2H), 4.27 (t, J = 7, 00 Hz, 2H), 2.10 (d, J = 6.34 Hz, 6H), 1.82-1.95 (m, 4H), 1.30 (td, J = 7.03, 3.68 Hz, 6H); LCMS (LCMS Method D): retention time = 0.78 min, [M + H] + = 695.4.

Example 58 (E) -1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) but -2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (2-hydroxyethoxy) -1Hbenzo Shimtsdazole-5-carboxamide

Example 58 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to (E) -1- (4- (5-carbamoyl2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo (N-imidazol-1-yl) but- 2-en-1-yl) -2- (1-ethyl-3 methyl-1H-pyrazole-5-carboxamido) -7- (2-methoxyethoxy) -1H-benzoHimidazole-5-carboxamide (300 mg,

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0.400 mmol) in DCM (5 ml) at 0 ° C was added BBr ₃ (501 mg, 2.00 mmol). After 3 hours, the reaction was quenched with water (5 ml) and the resulting solid was collected by filtration. This solid was purified by preparative HPLC to afford the title compound (21 mg, 0.029 mmol, 7% yield) as a gray solid. ¹ H NMR (400 MHz, DMSO-de) δ ppm 12.83 (s, 2H), 7.97 (d, J = 12.2 Hz, 3H), 7.62-7.78 (m, 2H) , 7.29-7.50 (m, 4H), 6.54 (d, J = 14.1 Hz, 2H), 5.99 (s, 1H), 5.86 (s, 1H), 4, 99 (s, 3H) s, 2H), 4.53 (d, J = 6, 8 Hz, 4H), 4.07 (s, 2H), 3.63 (s, 2H), 2.11 (s , 6H), 1.27 (s, 6H); LCMS (LCMS Method A): retention time = 1.333 min, [M + H] + = 737.1.

Example 59 (E) -1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -7-ethoxy-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo-i-imidazole-5-carboxamide

Example 59 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of the preparation is presented: HATU (171 mg, 0.450 mmol) and DIPEA were added to 1-ethyl-3-methyl-1Hpyrazole-5-carboxylic acid (58 mg, 0.38 mmol) in NMP (3 ml) at room temperature. 0.14 ml, 0.77 mmol). After 15 min, (E) -2-amino-1- (4- (2amino-5-carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7-ethoxy- 1H-benzo ^] imidazole-5-carboxamide (65 mg, 0.15 mmol) and the mixture was heated to 60 ° C. After 16 hours, water was added and the resulting solid was collected by filtration. This material was purified by preparative HPLC to afford the title compound (35 mg, 0.047 mmol, 33% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d) δ ppm 12.85 (s, 2H), 7.97 (d, J = 11.7 Hz, 3H), 7.72 (d, J = 8.4 Hz , 1H), 7.64 (s, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.32 (d, J = 12.3, 3H), 6.55 (s, 2H), 5.91-6.02 (m, 1H), 5.78 (cc, J = 13.3, 7.7 Hz, 1H), 4.94 (d, J = 4.7 Hz, 2 Hz) H), 4.83 (d, J = 4.9 Hz, 2H), 4.50-4.57 (m, 4H), 3.99-4.06 (s, 2H), 2.12 (d, J = 3.8 Hz, 6H), 1.28 (dd, J = 12.7, 6.9 Hz, 6H), 1.18 (t, J = 6.9 Hz, 3H); LCMS (LCMS Method A): retention time = 1.382 min, [M + H] + = 721.2.

Example 60 (E) -7-Bromo-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [d] imidazol-1-yl ) but-2-en-1-yl) -2- (1-e-tyl-3-methyl-1H-pyrazole-5 -carboxamido) - 1H-benzo [d] imidazole-5-carboxamide

Example 60 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: 1-ethyl-3-methyl-1Hpyrazole-5-carboxylic acid (1.12 g, 7.28 mmol), (E) -2-amino-1- (4- (2-amino-5 -carbamoyl-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) 7-bromo-1H-benzo ^] imidazole-5-carboxamide (1.6 g, 3.3 mmol), HATU ( 3.14 g, 8.28 mmol) and triethylamine (1.01 g, 9.93 mmol) were heated to 60 ° C in DMF (30 ml). After 12 hours, water (5 ml) was added and the resulting solid was collected by filtration. This material was purified by preparative HPLC to afford the title compound (700 mg, 0.926 mmol, 28% yield) as a brown solid. ¹ H NMR (400 MHz, DMSO-dQ δ ppm 13.05 (brs, 1H), 12.80 (s, 1H), 8.06 (s, 1H), 7.91-8.00 (m , 4H), 7.72 (d, J = 8 Hz, 1H), 7.43-7.47 (m, 2H), 7.33 (s, 1H), 6.54 (d, J = 4 Hz , 2H), 5.95-6.05 (m, 1H), 5.60-5.70 (m, 1H), 5.06-5.13 (m, 2H), 4.75-4.81 (m, 2H), 4.45-4.61 (m, 4H), 2.12 (d, J = 4 Hz, 6H), 1.18-1.35 (m, 6H); LC-MS ( LCMS Method A): retention time = 1.367 min, [M + H] + = 755.1.

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Example 61 (E) -7- (Aminomethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1H-benzo ^] imidazole-1 -yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazole-5-carboxamide trifluoroacetate

Example 61 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to (E) -tert-butyl ((5carbamoyl-1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5 -carboxamido) -1 H -benzoO | imidazol-1 yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoO | imidazol-7-yl) methyl ) carbamate (410 mg, 0.509 mmol) in MeOH (10 ml) at room temperature was added 36.5% HCl (0.5 ml,

0.51 mmol). The reaction mixture was warmed to 40 ° C and concentrated after 3 h. The residue was purified by preparative HPLC to give the title compound (9 mg, 11 μmol, 2% yield). 1H NMR (400 MHz, DMSOD ₆ ) δ ppm 13.07-12.88 (m, 1H), 8.34 (s, 3H), 8.00 (d, J = 7.3 Hz, 2H), 7 , 95 (s, 1H), 7.88 (s, 1H), 7.74 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 8.4, 2H), 7, 35 (s, 1H), 6.56 (d, J = 15.5 Hz, 2H), 5.92 (d, J = 15.9 Hz, 1H), 5.58 (d, J = 16.0 Hz, 1 H), 5.23-5.29 (m, 2H), 5.00 (s, 2H), 4.82 (s, 2H), 4.53 (dd, J = 13.6, 6 , 6 Hz, 4H), 4.15 (d, 4.6 Hz, 2H), 2.12 (d, J = 3.9 Hz, 6H), 1.28 (d, J = 4.2 Hz, 6H); LCMS (LCMS Method A): retention time = 1.183 min, [M + H] + = 706.3.

Example 62 (E) -8-Ethyl-1,26-dimethoxy-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28 , 31 dodecahydrobenzo [4,5] imidazo [1,2-a] -benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3 , 6,15,17] pentaazacyclogenaicosine-3,24-dicarboxamide-bis-trifluoroacetate

Example 62 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to a solution of 4- (5- (5carboxy-3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (37.5 mg, 0.108 mmol), HATU (101 mg, 0.267 mmol) and TEA (86 μl, 0.62 mmol) in DMF (4.1 ml) was added (E) 4,4 '- (but-2-en-1 , 4-diylbis (azandiyl) bis (3-amino-5-methoxybenzamide) dihydrochloride (50 mg, 0.10 mmol) The reaction mixture was heated to 100 ° C and after 3 h water was added.The resulting solid was collected by filtration and purified by HPLC to give the title compound (1 mg, 1 μmol, 1% yield, low yield in part due to malfunctioning injection port during HPLC purification) 1H NMR (400 MHz, MeOH-d4) δ ppm 7.68 (s, 1H), 7.64 (s, 1H), 7.37 (d, J = 4.56 Hz, 2H), 6.63 (s, 1H), 5.76 (d, J = 12.67 Hz, 2H), 5.05 (d, J = 12.93 Hz, 4H), 4.70 (br s, 2H), 4.46-4.57 (m, 2H), 3.72 (d, J = 8.36 Hz, 6H), 2.81 (d, J = 6.59 Hz, 2H), 2.26 (s, 3H), 2.20 (s, 3H), 1.86 (br s , 2H), 1.58 (br s, 2H), 0.890.96 (m, 5H); LC-MS (LC-MS method D): retention time = 0.91 min, [M / 2 + H] ⁺ = 389.5.

Example 63 8-Ethyl-1,26-bis (3-hydroxypropoxy) -10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,

15,20,21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1е: 4 ' , 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-3,24-dicarboxamide dihydrochloride

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Example 63 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to 1,26-bis (3 - ((tert-butyldimethylsilyl) oxy) propoxy) -8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13 , 14,15,20,21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1 -e: 4 ', 3'-1] [1,3,6, 15,17] pentaazacyclogeneicosin-3,2 4-dicarboxamide (18 mg, 0.016 mmol) in THF (0.5 ml) at 0 ° C was added 4N HCl solution (0.025 ml, 0.099 mmol). After 60 min, the resulting precipitate was collected by filtration and washed with EtOAc to give the title compound (15 mg, 0.016 mmol, 97% yield). 1H NMR (400 MHz, DMSO-4 _b ) δ ppm 7.95-8.08 (m, 2H), 7.64-7.70 (m, 2H), 7.29-7.42 (m, 4H ), 6.51-6.58 (m, 1H), 4.63-4.72 (m, 2H), 4.31-4.50 (m, 6H), 4.11-4.20 (m , 4H), 3.42-3.48 (m, 4H), 2.74-2.85 (m, 2H), 2.15 (s, 3H), 2.10 (s, 3H), 1 , 78-1.91 (m, 6H), 1.64-1.74 (m, 4H), 1.48-1.58 (m, 2H), 1.37-1.47 (m, 2H) , 1.26-1.32 (m, 3H); LCMS (LCMS Method D): retention time = 0.82 min, [M + H] + = 867.5.

Example 64 (29R, 30R) -8-Etuyl-29,30-dihydroxy-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20.21 , 28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3' 1] [1,3,6,15,17] pentaazacycloeneicosin-3,24-dicarboxamide

Example 64 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: ^ aR ^ aR) ^^ ™ 10,18,30,30-tetramethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28 , 28а, 31а, 32-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1 ] [1,3,6,15,17] pentaazacyclogenaicosin-3,24-dicarboxamide (500 mg, 0.632 mmol), formic acid (15 ml, 391 mmol) and water (1.5 ml) were stirred at 25 ° C. After 48 h, the mixture was concentrated and the residue was purified by preparative HPLC (Gemini-C18 column, silica gel 5 μm, diameter 21 mm, length 150 mm), eluting with a mixture of 10-30% water in MeCN (0.1% formic acid) to obtain the the title compound (7.5 mg, 9.5 μmol, 1.5% yield) as a white solid. ¹ H NMR (400 MHz, MeOH-d ₄ ) δ ppm 8.01 (d, J = 13, l Hz, 2H), 7.81-7.93 (m, 2H), 7.63-7.76 (m, 2H), 6.66 (s, 1H), 4.83 (s, 1H), 4.62-4.74 (m, 1H), 4.44-4.63 (m, 6H), 4.12-4.33 (m, 2H), 2.90-3.00 (m, 1H), 2.71-2.82 (m, 1H), 2.23 (d, J = 18.2 Hz, 6H), 1.84-2.00 (m, 2H), 1.61-1.73 (m, 2H), 1.37-1.47 (m, 2H), 1.38 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method A): retention time = 1.295 min, [M + H] + = 751.2.

Example 65 8-Ethyl-10,13,13,13,18-tetramethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31 tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6, 15,17] pentaazacyclogeneicosin-3,24-dicarboxamide

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Example 65 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to 4- (5- (5-carboxy-3methyl-1H-pyrazol-1-yl) -3,3-dimethylpentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (192 mg, 0.510 mmol) in NMP (10 ml) was added HATU (194 mg, 0.510 mmol). The reaction mixture was heated to 40 ° C, and 1,1 '- (propane-1,3-diyl) bis (2-amino-1H-benzo Schimidazole-5-carboxamide) (200 mg, 0.510 mmol) was added. After heating overnight, the reaction mixture was purified by preparative HPLC to give the title compound (13 mg, 0.017 mmol, 3.4% yield). 1H NMR (400 MHz, MeOH-d ₄ ) δ ppm 8.01 (d, J = 8.4 Hz, 2 H), 7.85 (d, J = 8.4 Hz), 7.51 (t, J = 8.4 Hz, 2H), 6.65 (s, 1H), 4.73-4.77 (m, 2H), 4.45-4.53 m, 2H), 4.28-4, 33 (m, 4H), 2.78-2.87 (m, 2H), 2.29 (d, J = 11.6 Hz, 6H), 1.98 (br s, 4H) 1.88 ( t, J = 7.6 Hz, 2H), 1.45-1.54 (m, 2H), 1.27-1.39 (m, 5H), 1.07 (s, 6H); LCMS (LCMS Method A): retention time = 1.397 min, [M + H] + = 747.3.

Example 66 8-Ethyl-12,13-dihydroxy-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,

29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogenaicosin-3,24-dicarboxamide

Example 66 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to 8-ethyl-10,18-dimethyl7,20-dioxo-6,7,8,11,14,15,20,21,28,29,30,31-dodecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacycloneicosin-3 , 24-dicarboxamide (350 mg, 0.488 mmol) and NMO (114 mg, 0.977 mmol) in tert-BiOH (9 ml) and water (3 ml) was added osmium tetroxide (7.7 μl, 0.024 mmol). After 4 h at 25 ° C, the reaction was quenched with Na ₂ SO ₃ and the reaction mixture was diluted with DCM (50 ml). The mixture was washed with water (30 ml) and brine (30 ml), and the organic layer was dried over Na ₂ SO ₄ and concentrated. The residue was purified by preparative HPLC (Gemini-C18 column, silica gel 5 μm, diameter 21 mm, 150 mm) eluting with 20-40% MeCN in water (0.1% TFA) to afford the title compound (11 mg, 0.015 mmol, yield 3.0%) as a white solid. ¹ H NMR (400 MHz, MeOH-d ₄ ) δ ppm 8.14 (s, 1H), 7.94 (dd, J = 27.0, 8.4 Hz, 2H), 7.68 (dd, J = 37.9, 8.5 Hz, 2H), 6.68 (s, 1H), 5.29-5.41 (m, 1H), 4.38-4.91 (m, 6H), 4, 20-4.29 (m, 1H), 3.74 (t, J = 12.4 Hz, 2H) dd, J = 14.1, 10.2 Hz, 1H), 2.95 (d, J = 11.2 Hz, 1H), 2.38-2.49 (m, 1H), 2.29 (s, 1H), 2.29 (s, 3H), 2.27 (s, 3H), 2, 04-2.15 (m, 3H), 1.92-1.99 (m, 1H), 1.45 (t, J = 8 Hz, 3H); LCMS (LCMS Method A): retention time = 1.270 min, [M + H] + = 750.9.

Example 67. (1R, 39R) -14-Ethyl-16,25-dimethyl-12,28-dioxo-2,9,11,14,15,23,24,29,31,38-decaazoctacyclo [37.2.2.0 ² , ¹⁰ , 0 ³ , ⁸ , 0 ¹³ , ¹⁷ , 0 ²³ , ²⁷ '0 ³⁰ , ³⁸ , 0 ³² , ³⁷ ] tritetrakonta-3,5,7,9,13 (17), 15,24,26, 30,32,34,36 dodecaene-6,34-dicarboxamide

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Example 67 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to 4- (5- (5-carboxy-3methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (483 mg, 1 , 39 mmol) in DMF (10 ml) at 25 ° C was added HATU (1.10 g, 2.89 mmol) and DIEA (1.01 ml, 5.78 mmol). After 4 h, 1,1 '- ((^^) cyclohexane-1,4-diyl) bis (2-amino-1H-benzo-imidazole-5-carboxamide) (500 mg, 1.16 mmol) was added and the reaction mixture was stirred at 80 ° C for 16 h. Water was added and a brown solid appeared and collected. The solid was purified twice to give the product (3 mg, 4 μmol, 0.3% yield). ¹ H NMR (400 MHz, DMSO-de) δ ppm 13.04 (s, 2H), 8.07 (d, J = 8.2 Hz, 6H), 7.88 (d, J = 8.2 Hz, 2H), 7.39 (s, 2H), 6.58 (s, 1H), 4.95 (br s, 2H), 4.69 (br s, 2H), 4.48 (br s, 2H), 2.60-2.85 (m, 6H), 1.99-2.24 (m, 10H), 1.80-2.01 (m, 3H), 1.49-1 , 65 (m, 4H), 1.34 (br s, 4H); LCMS (LCMS Method A): retention time = 1.413 min, [M + H] + = 745.3.

Example 68. 1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-isopropoxy-1H-benzo Schimidazole 5-carboxamide

Example 68 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is presented: DIEA (0.489 ml, 2.80 mmol) was added to 1-ethyl-3-methyl-1Hpyrazole-5-carboxylic acid (215 mg, 1.40 mmol) in DMF (5 ml) at 25 ° С and HATU (638 mg, 1.68 mmol). After 30 min, 2-amino-1- (4- (2-amino-5-carbamoyl1H-benzo ^] imidazol-1-yl) butyl) -7-isopropoxy-1H-benzo Schimidazole-5-carboxamide (260 mg, 0.560 mmol). After 16 h, water was added and the resulting precipitate was collected by filtration and washed with MeOH to give the title compound (55 mg, 0.075 mmol, 13% yield) as a brown solid. 1H NMR (400 MHz, DMSOC ₆ ) δ ppm 12.80 (d, J = 12.5 Hz, 2H), 7.88-8.01 (m, 3H), 7.75 (d, J = 7, 8 Hz, 1H), 7.55-7.60 (m, 2H), 7.30 (brs, 3H), 6.59 (s, 2H), 4.74-4.84 (m, 1H ), 4.52-4.63 (m, 4H), 4.294.36 (m, 4H), 2.11 (d, J = 6.5 Hz, 6H), 1.77-1.94 (m, 4H), 0.98-1.44 (m, 12H); LCMS (LCMS Method A): retention time = 1.428 min, [M + H] + = 737.2.

Example 69. 1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -7 - ethoxy-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazole-5carboxamide

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Example 69 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: DIEA (0.550 ml, 3.15 mmol) was added to 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (191 mg, 1.24 mmol) in DMF (10 ml) at 25 ° С and HATU (600 mg, 1.58 mmol). After 30 minutes, 2-amino-1- (4- (2-amino-5carbamoyl-1H-benzo [d] imidazol-1 -yl) butyl) -7-ethoxy-1H-benzo [d] imidazole-5-carboxamide (270 mg, 0.599 mmol). After 16 h, water was added and the resulting precipitate was collected by filtration and washed with MeOH to give the title compound (28 mg, 0.039 mmol, 6% yield). 1H NMR (400 MHz, DMSO <) δ ppm 8.00 (s, 2H), 7.94 (s, 1H), 7.74 (d, J = 8.4 Hz, 1 H), 7.59 ( s, 1H), 7.50 (d, J = 8.4 Hz, 1 H), 7.35 (s, 2 H), 7.30 (s, 1H), 6.59 (s, 2 H) , 4.50-4.64 (m, 4 H), 4.31 (d, J = 36.9 Hz, 4H), 4.12 (d, J = 7.0 Hz, 2 H), 2, 10 (d, J = 1.7 Hz, 6 H), 1.86 (s, 4 H), 1.29 (dd, J = 7.7, 6.6 Hz, 6 H), 1.24 ( d, J = 7.0 Hz, 3 H); LCMS (LCMS Method A): retention time = 1.426 min, [M + H] + = 723.2.

Example 70. 1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methyl-1H-benzo Schimidazole-5carboxamide

Example 70 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: DIEA (0.900 ml, 5.15 mmol) was added to 1-ethyl-3-methyl-1Hpyrazole-5-carboxylic acid (331 mg, 2.15 mmol) in NMP (10 ml) at room temperature and HATU (979 mg, 2.58 mmol). After 30 min, 2-amino-1- (4- (2amino-5-carbamoyl-1H-benzo [d] imidazol-1-yl) butyl) -7-methyl-1H-benzo [d] imidazole-5-carboxamide was added dihydrobromide (500 mg, 0.859 mmol) and the reaction mixture was heated to 60 ° C overnight. Water was added and the resulting precipitate was collected by filtration and purified by preparative HPLC to give the title compound (45 mg, 0.065 mmol, 7.6% yield) as a white solid. 1H NMR (400 MgtDMSOO δ ppm 12.85 (s, 2H), 7.98 (s, 2H), 7.91 (s, 1H) s), 7.77 (d, J = 8.4 Hz, 1H ), 7.54-7.64 (m, 2H), 7.32 (d, J = 9.9 Hz, 2H), 6.60 (d, J = 15.5 Hz, 2H), 4.56 (dd, J = 10.8, 7.1 Hz, 4H), 4.39 (s, 2H), 4.29 (s, 2H), 2.62 (s, 3H), 2.09 (d, 13.4 Hz, 6H), 1.89 (d, J = 22, l Hz, 4H), 1.30 (q, J = 7.3 Hz, 6H); LCMS (LCMS Method A): retention time = 1.341 min, [M + H] + = 693.3.

Example 71. 1- (4- (5-Carbamoyl-2 - (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (morpholinomethyl) -1H-benzo H | imidazole-5-carboxamide

- 131 036435

Example 71 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of the preparation is presented: DIEA (0.777 ml, 4.45 mmol) was added to 1-ethyl-3-methyl-1Hpyrazole-5-carboxylic acid (302 mg, 1.96 mmol) in NMP (3 ml) at room temperature and HATU (846 mg, 2.2-3 mmol). After 1 h, 2-amino-1- (4- (2-amino5-carbamoyl-1 H-benzo [b] imidazol-1 -yl) butyl) -7- (morpholinomethyl) -1 H-benzo [b] imidazole -5-carboxamide (450 mg, 0.890 mmol) and the reaction mixture was heated to 60 ° C overnight. Water was added and the resulting precipitate was collected by filtration to give the title compound (70 mg, 0.090 mmol, 10% yield) as a white solid. ¹ H NMR (400 MHz, DMSO-b ₆ ) δ ppm 12.99 (br s, 2H), 9.89 (br s, 2H), 7.99-8.11 (m, 5H), 7 , 87 (s, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.4 Hz, 2H), 7.35 (s, 1H), 6 , 66 (s, 1H), 6.62 (s, 1H), 4.53-4.61 (m, 5H), 4.47 (s, 2H), 4.26 (s, 2H), 3, 92 (brs, 2H), 3.60 (brs, 2H), 3.36 (brs, 2H), 2.10 (s, 3H), 2.07 (s, 3H), 1.83 (d, J = 22.7 Hz, 4H), 1.23-1.45 (m, 6H); LCMS (LCMS Method A): retention time = 1.255 min, [M + H] + = 778.3.

Example 72 4- (2- (Dimethylamino) acetamido) -8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21, 28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1е: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-2 4-carboxamide

Example 72 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to 4-amino-8-ethyl-10.18dimethyl-7.20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30, 31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3 , 6,15,17] pentaazacyclogenaicosin-24-carboxamide (200 mg, 0.290 mmol), 2- (dimethylamino) acetic acid (119 mg, 1.188 mmol), HOBt (44.3 mg, 0.290 mmol), DIEA (0.405 ml , 2.32 mmol) and DMAP (17.7 mg, 0.145 mmol) in DMF (5 ml) at 60 ° C were added 10 portions of HATU (661 mg, 1.7 4 mmol) at intervals of 30 min. The reaction mixture was cooled to room temperature, diluted with water, and the resulting solid was collected by filtration, washed with water and air dried. The solid was then purified on silica gel (Combiflash Rf 120 g column, 85 mL / min) eluting with 0-20% MeOH in DCM to give the title compound (205 mg, 0.251 mmol, 87% yield) as an off-grade white solid. 1H NMR (400 MHz, DMSO-b ₆ ) δ ppm 12.90 (br s, 1H), 12.55 (br s, 1H), 10.37 (br s, 1H), 8.02 ( s, 1H), 7.99 (br s, 1H), 7.83 (dd, J = 8.6, 1.5 Hz, 1H), 7.66 (d, J = 8.6 Hz, 1H ), 7.58 (d, J = 7.8 Hz, 1H) d, J = 7.6 Hz, 2H), 7.227.29 (m, 1H), 6.57 (s, 1H), 4.69 -4.80 (m, 2H), 4.47 (d, J = 7.1 Hz, 2H), 4.14-4.34 (m, 4H), 3.25 (s, 2H), 2, 81 (bs, 2H), 2.33 (s, 6H), 2.16 (s, 3H), 2.08 (s, 3H), 1.91 (bs, 4H), 1.76 -1.86 (m, 2H), 1.48 (d, J = 5.9 Hz, 2H), 1.38 (d, J = 5.1 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.82 min, [M + H] + = 776.7.

- 132 036435

Example 73 7- (Aminomethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1HbenzoH | imidazol-1-yl) butyl) - 2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoO | imidazole-5carboxamide

Example 73 can be prepared in accordance with method 9 with modifications known to the person skilled in the art. The last stage of obtaining is given: to tert-butyl ((5-carbamoyl-1- (4- (5carbamoyl-2- (1-ethyl-3-methyl-III-pyrazole-5-carboxamido) 1H-benzo ^ imidazole-1 - yl) butyl) -2- (1-ethyl-3 methyl-1H-pyrazole-5-carboxamido) -1H-benzoH-imidazol-7-yl) methyl) carbamate (200 mg, 0.248 mmol) in MeOH (10 ml) at room temperature temperature was added 12 M HCl solution (0.5 ml, 16.5 mmol). After stirring overnight, the mixture was concentrated and purified by preparative HPLC to give the title compound (50 mg, 0.071 mmol, 29% yield) as an off-white solid. '11 NMR (400 MHz, gMCO-d6) δ ppm 8.21 (s, 1H), 7.89-8.01 (m, 5H), 7.76 (d, J = 6.5 Hz, 2H) , 7.56 (d, J = 8.4 Hz, 1H), 7.33 (s, 2H) 6.59 (d, J = 12.7 Hz, 2H), 4.47-4.42 (m , 6H), 4.28 (s, 2H), 4.07 (s, 2H), 2.08 (d, J = 10.8 Hz, 6H), 1.89 (d, J = 17.6 Hz , 4H), 1.29 (q, J = 6.9 Hz, 6H); LCMS (LCMS Method A): retention time = 1.176 min, [M + H] + = 708.3.

Example 74 (E) -1,26-Dibromo-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28 , 31 dodecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3, 6,15,17] pentaazacyclogeneicosin-3,24-dicarboxamide

Example 74 can be prepared according to method 6 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: c (E) -1,1 '- (but-2-ene-1,4diyl) bis (2-amino-7-bromo-1H-benzo-imidazole-5-carboxamide) (500 mg, 0.889 mmol ), 4- (5- (5-carboxy3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (310 mg, 0.889 mmol) and HATU (1.01 g, 2.67 mmol) in DMF (15 ml) was added DIEA (0.466 ml, 2.67 mmol) and the reaction mixture was heated to 90 ° C. After 1.5 hours, the mixture was poured into water and the resulting solid was collected by filtration. The crude material was purified on silica gel eluting with DCM: MeOH 5: 1 with 0.1% aqueous NH ₃ solution to give the title compound (250 mg, 0.245 mmol, 24% yield) as a brown solid. 'H NMR (400 MHz, DMSOA,) δ ppm 13.04 (brs, 2H), 8.02-8.10 (m, 4H), 7.92 (s, 2H), 7.47 ( s, 2H), 6.51 (s, 1H), 5.72-5.78 (m, 2H), 5.01-5.25 (m, 4H), 4.52-4.66 (m, 2H) 4.39-4.50 (m, 2H), 2.71-2.77 (m, 2H), 2.17 (s, 3H), 2.10 (s, 3H), 1.62- 1.76 (m, 2H), 1.45-1.54 (s, m, 2H), 1.24-1.40 (m, 5H); LCMS (LCMS Method A): retention time = 1.448 min, [M + H] + = 873.2.

Example 75.

Example 75 is a mixture of two isomers 28-ethyl-17,26-dimethyl-14,30-dioxo 4,11,13,18,19,27,28,31,33,40-decazoctacyclo [42,3,1,0 ⁴ , ¹² , 0 ⁵ , ¹⁰ , 0 ¹⁵ , ¹⁹ , 0 ²⁵ , ²⁹ , 0 ³² , ⁴⁰ , 0 ³⁴ , ³⁹ ] octatetraconte-1 (48), 5,7,9,11,15,17,25 ( 29), 26,32,34,36,38,44,46-pentadecaen-8,36-dicarboxamide and 16-ethyl-18,27-dimethyl14,30-dioxo-4,11,13,16,17,25 , 26,31,33,40-decaazoctacyclo [42,3,1,0 ⁴ , ¹² , 0 ⁵ , ¹⁰ , 0 ¹⁵ , ¹⁹ , 0 ²⁵ , 0 ²⁹ , 0 ³² , ⁴ 0.0 ³⁴ , ³⁹ ] ok - 133 036435 tatetrakonta-1 (48), 5,7,9,11,15 (19), 17,26,28,32,34,36,38,44,46-pentadecaen-8,36-dicarboxamide

Example 75 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to 4- (5- (5-carboxy-3methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (140 mg, 0.403 mmol) in NMP (10 ml) was added at 40 ° C HATU (153 mg, 0.403 mmol) and 2-amino-1- (3- (3- (2- (2-amino-5carbamoyl-1H-benzo [d] imidazol-1-yl) ethyl) phenyl) propyl) -1 H-benzo [d] imidazole-5-carboxamide (200 mg, 0.403 mmol). After stirring overnight, the reaction mixture was purified by preparative HPLC to give the compounds as a mixture (2 mg, 2 μmol, 0.6% yield). ¹ H NMR (400 MHz, MeOH-d ₄ ) δ ppm 7.97 (s, 1H), 7.84-7.91 (m, 2H), 7.73 (d, J = 8.4 Hz, 1H ), 7.51 (d, J = 8.4 Hz, 1H), 7.20 (d, J = 8.4 Hz, 1H), 6.936.98 (m, 2H), 6.81-6.89 (m, 2H), 6.64 (s, 1H), 4.66-4.72 (m, 2H), 4.49-4.57 (m, 2H), 4.40-4.47 (m , 2H), 4.25-4.31 (m, 2H), 2.96-3.02 (m, 2H), 2.78-2.84 (m, 2H), 2.60-2.66 (m, 2H), 2.28 (s, 3H), 2.18, 3H), 1.83-1.92 (m, 2H), 1.56-1.64 (m, 2H), 1, 29-1.39 (m, 7H); LCMS (LCMS Method A): retention time = 1.428 min, [M + H] + = 809.3.

Example 76 4-Amino-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29,30,31 -tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3, 6,15,17] pentaazacyclogeneicosine-24-carboxamide

Example 76 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to pure tert-butyl (24carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28 , 29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1 ] [1,3,6,15,17] pentaazacyclogenaicosin-4-yl) carbamate (2.85 g, 3.60 mmol) was slowly added TFA (10 ml, 130 mmol). After 30 minutes, the reaction mixture was concentrated. The residue was suspended in water, treated with NaHCO ₃ to basic pH, filtered and washed with water. This material was purified on silica gel (120 g column) eluting with 0-20% MeOH in DCM (2.35 g, 3.23 mmol, 90% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSO ^) δ ppm 12.90 (s, 1H), 12.36 (s, 1H) 8.02 (d, J = 1.2 Hz, 1H), 7.99 (br s, 1H), 7.83 (dd, J = 8.4, 1.3 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.34 (br s, 1H), 6.91-7.03 (m, 1H), 6.70 (d, J = 7.8 Hz, 1H), 6.57 (s, 1H), 6.48 (d, 7.8 Hz, 1H), 5.92 (s, 2H), 4.74 (t, J = 7.1 Hz, 2H), 4.48 (q, J = 7.1 Hz, 2H), 4.26 ( brs, 2H), 4.14 (brs, 2H), 2.74-2.88 (m, 2H), 2.16 (s, 3H), 2.08 (s, 3H), 1 , 72-1.96 (m, 6H), 1.48 (d, J = 5.4 Hz, 2H), 1.38 (d, J = 5.6 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.91 min, [M + H] ⁺ = 691.5.

Example 77. 1 - (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2 - (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -7- (2-hydroxyethoxy) -1H-benzo [d] imidazole-5-carboxamide

- 134 036435

Example 77 can be prepared according to Method 4 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to a suspension of 1- (4- (5carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo (imidazol-1-yl) butyl) -2- (1-ethyl3-methyl-1H-pyrazole-5-carboxamido) -7- (2-methoxyethoxy) -1H-benzoHimidazole-5-carboxamide (100 mg, 0.133 mmol) in DCM (5 ml) at room temperature, was added drops of BBr ₃ (0.126 ml, 1.33 mmol). After 2 h, the reaction was quenched with MeOH, the reaction mixture was concentrated and the residue was purified by preparative HPLC to afford the title compound (15 mg, 0.020 mmol, 15% yield) as a white solid. ¹ H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.81 (br s, 2H), 7.93-8.01 (m, 3H), 7.75 (d, J = 8.8 Hz, 1H ), 7.63 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.31-7.38 (m, 3H), 6.59 (d, J = 11, 6 Hz, 2H), 4.94 (brs, 1H), 4.54-4.61 (m, 4H), 4.40-4.49 (m, 2H), 4.21-4.32 (m, 2H), 4.12-4.20 (m, 2H), 3.70-3.76 (m, 2H), 2.08 (d, J = 5.6 Hz, 6H), 1, 77-1.91 (m, 4H), 1.22-1.35 (m, 6H); LCMS (LCMS Method A): retention time = 1.290 min, [M + H] + = 739.2.

Example 78 (E) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1- (4- (2- (1-ethyl-3-methyl-1H-pyrazole 5-carboxamido) -7-methoxy-1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -7-methoxy-1H-benzoSimidazole 5-carboxamide bis-trifluoroacetate

Example 78 can be prepared according to method 11 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to (E) -2-amino-1- (4- (2amino-7-methoxy-1H-benzo-thymidazol-1-yl) but-2-en-1-yl) -7-methoxy-1H -benzoHimidazole-5-carboxamide (250 mg, 0.178 mmol), HATU (203 mg, 0.534 mmol), HOBt (40.9 mg, 0.267 mmol) and 1-ethyl-3methyl-1H-pyrazole-5-carboxylic acid (82 mg, 0.53 mmol) in DMF (3 ml) was added TEA (0.149 ml, 1.07 mmol). After stirring over the weekend, the reaction mixture was passed through a syringe filter and purified by reverse phase HPLC (Gilson column, Sunfire Prep C18 OBD, 5 μm, 30 mm x 100 mm), eluting with 20-90% MeCN in water (0.1% TFA) with obtaining the title compound (18 mg, 0.020 mmol, 11% yield) 1H NMR (400 MHz, MeOH-d ₄ ) δ ppm 7.60 (d, J = 1.27 Hz, 1H), 7.33 ( d, J = 1.27 Hz, 1H), 7.26 (s, 1H) J = 8.11 Hz, 1H), 6.87 (d, J = 8.11 Hz, 1H), 6.58 ( s, 1H), 6.54-6.57 (m, 1H), 5.85-5.93 (m, 2H), 5.14 (br s, 2H), 5.06 (d, J = 3.80 Hz, 2H), 4.59 (dd, J = 13.81, 6.97 Hz, 4H), 3.79 (s, 6H), 2.20 (d, J = 1.77 Hz, 6H) 1.36 (td, J = 7.16, 2.91 Hz, 6H); LCMS (LCMS Method D): retention time = 1.02 min, [M + H] + = 694.5.

Example 79 8-4- (2-Aminoacetamido) -8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28 , 29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1 ] [1,3,6,15,17] pentaazacyclogenaicosin-24-carboxamide

- 135 036435

Example 79 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to tert-butyl (2 - ((24carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20, 21,28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3 '-1] [1,3,6,15,17] pentaazacycloneicosin-4-yl) amino) -2-oxoethyl) carbamate (350 mg, 0.413 mmol) was slowly added TFA (3.00 ml, 38.9 mmol) ... After 30 minutes, the reaction mixture was concentrated. The resulting residue was suspended in water and treated with NaHCO ₃ to basic pH. The solid was collected by filtration and washed with water. This material was purified on silica gel (80 g column) eluting with 0-20% (2N NH ₃ in MeOH) in DCM to give the title compound (270 mg, 0.333 mmol, 83% yield) as an off-white solid matter. ¹ H NMR (400 MHz, DMSOD6) δ ppm 8.03 (d, J = 1.2 Hz, 1H), 7.99 (brs, 1H), 7.83 (dd, J = 8.3, 1.5 Hz, 1H), 7.58-7.69 (m, 2H), 7.34 (d, J = 7.3 Hz, 2H), 7.207.31 (m, 1H), 6.57 ( s, 1H), 4.74 (t, J = 7.1 Hz, 2 H), 4.47 (q, J = 7.1 Hz, 2H), 4.14-4.34 (m, 4H) , 3.54 (s, 2H), 2.76-2.85 (m, 2H), 2.16, 3H), 2.09 (s, 3H), 1.91 (br s, 4H), 1.82 (d, J = 6.6 Hz, 2H), 1.48 (d, J = 5.6 Hz, 2H), 1.38 (d, J = 4.9 Hz, 2H), 1, 29 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.78 min, [M + H] + = 748.6.

Example 80. 8-Ethyl-10,17-dimethyl-7,19-dioxo-7,8,11,12,13,14,19,20,27,28,29,30-dodecahydro-6Hbenzo [4.5 ] imigazo [1,2-a] benzo [4,5] imigazo [2,1-o] dipyrazolo [5,1-e: 4 ', 3'-k] [1,3,6,14,16] pentaazacyclogeneicosin-3,2 3-dicarboxamide

Example 80 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: a mixture of 4- (4- (5-carboxy-3methyl-1H-pyrazol-1-yl) butyl) -1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (165 mg, 0.492 mmol), 1,1 '- (butane-1,4-diyl) bis (2-amino-1H-benzo-imidazole-5-carboxamide) (200 mg, 0.492 mmol), HATU (561 mg, 1.48 mmol) and DIEA (382 mg, 2.95 mmol) in NMP (8 ml) was stirred at 120 ° C for 18 h. The mixture was concentrated and the residue was purified on silica gel eluting with DCM: MeOH: NH ₄ OH (aq) 80:20 :one. The partially pure product was purified by preparative HPLC to give the title compound (7 mg, 10 μmol, 2% yield). ¹ H NMR (400 MHz, DMSOD6) δ ppm 12.83 (br s, 2H), 7.90-8.01 (m, 4H), 7.79 (t, J = 6 Hz, 2H), 7.52 (m, 2H), 2H), 4.25 (br. C, 4H), 2.90-3.01 (m, 2H), 2.17 (s, 3H), 2.08 (s , 3H), 1.78-1.89 (br m, 6H), 1.53-1.57 (m, 2H), 1.30 (t, J = 6 Hz, 3H); LCMS (LCMS Method A): retention time = 1.316 min, [M + H] + = 704.7.

Example 81 8-Ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,14,15,20,21,28,29,30,31-dodecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacycloneicosin- 3.2 4-dicarboxamide

- 136 036435

Example 81 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to 4- (5- (5-carboxy-3methyl-1H-pyrazol-1-yl) pent-2-en-1-yl) -1-ethyl-3-methyl-1H-pyrazole-5 -carboxylic acid (320 mg, 0.924 mmol), 1, G- (butane-1,4-diyl) bis (2-amino-1H-benzoSimidazole-5-carboxamide) (488 mg, 1.2 0 mmol) and DIEA (0.4 84 ml, 2.7 7 mmol) in NMP (10 ml) was added HATU (87 8 mg, 2.31 mmol). The reaction mixture was stirred at 80 ° C for 16 h, then poured into Et ₂ O (50 ml). The formed precipitate was collected by filtration and washed with Et ₂ O and water. The crude product was purified by preparative HPLC (Gemini-C18 column, silica gel 5 μm, diameter 21 mm, length 150 mm) eluting with 20-40% MeCN in water (containing 0.1% TFA) to afford the title compound (11 mg, 15 μmol, 1.7% yield) as a white solid. ¹ H NMR (400 MHz, MeOH-d ₄ ) δ ppm 8.00 (s, 1H), 7.93 (s, 1 H), 7.81 (t, J = 9.7 Hz, 2H), 7 , 56-7.40 (m, 2H), 6.66 (s, 1H), 5.77 (d, J = 15.2 Hz, 1H) d, J = 15.5 Hz, 1H), 4, 78-4.85 (m, 1H), 4.68-4.52 (m, 2H), 4.37-4.22 (m, 4H), 3.62 (d, J = 5.8 Hz, 2 H), 2.82-2.49 (m, 2H), 2.23 (s, 3H), 2.13 (s, 3H), 2.09-1.93 (m, 5H), 1, 41 (t, J = 8.0 Hz, 3H); LCMS (LCMS Method A): retention time = 1.376 min, [M + H] + = 716.9.

Example 82. 35-Ethyl-5,37-dimethyl-8,33-dioxo-3,4,9,11,18,23,30,32,35,36-decaazoctacyclo [38,1,1,0 ³ , ⁷ , 0 ¹⁰ , ¹⁸ , 0 ¹² , ¹⁷ , 0 ²³ , ³¹ , 0 ²⁴ , ²⁹ , 0 ³⁴ , ³⁸ ] dotetrakonta-4,6,10,12,14,16,24,26,28,30,34 (38), 36-dodecaene-14,27-dicarboxamide

Example 82 can be prepared according to method 19 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to 4 - ((3 - ((5-carboxy-3methyl-1H-pyrazol-1-yl) methyl) cyclobutyl) methyl) -1-ethyl-3-methyl-1H pyrazole-5-carboxylic acid (175 mg, 0.486 mmol), 1, G- (butane-1,4-diyl) bis (2-amino-1H-benzoSimidazole-5-carboxamide) (220 mg, 0.541 mmol) and HATU (559 mg, 1, 4 7 mmol) in NMP (2 ml) at room temperature was slowly added

DIEA (0.514 ml, 2.94 mmol). The mixture was heated to 70 ° C for 4 h and poured into water. The crude product was collected by filtration and the filtrate was concentrated to obtain a second batch of crude product, which was washed with water. The combined crude product was purified by preparative HPLC (Gemini-C18 column, 5 μm silica gel, 21 mm diameter, 150 mm length) eluting with 3060% MeCN in water (containing 0.1% TFA) to afford the title compound as a white solid matter. 1H NMR (400 MHz, MeOH-d ₄ ) δ ppm 12.87 (s, 2H), 8.02 (t, J = 9.0 Hz, 4H), 7.83 (dd, J = 19.9, 8.4 Hz, 2 H), 7.65 (dd, J = 23.0, 8.2 Hz, 2H), 7.37 (d, J = 2.1 Hz, 2H), 6.52 (s , 1H), 4.64 (d, J = 5.3 Hz, 2H), 4.44 (m, 2H), 4.28 (s, 4H), 2.91 (d, J = 7.3 Hz , 2H), 2.47 (m, 2H), 2.14 (s, 3H), 3H), 1.96 (s, 6H), 1.53 (m, 2H), 1.28 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method A): retention time = 1.351 min, [M + H] + = 731.3.

Example 83 (E) -1- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoSHimidazol-1-yl) but-2-1 -yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoH-imidazole-5-carboxylic acid

- 137 036435

Example 83 can be prepared according to method 11 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: k (E) -methyl 1- (4- (5carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1H-benzo ^ imidazol-1-yl) but-2-en-1-yl) -2 (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5-carboxylate (1.26 mmol) in MeOH (10 ml), DMF (10 ml) and water (10 ml) at room temperature was added NaOH (0.729 g, 18.2 mmol). After 3 h, another portion of NaOH (0.729 g, 18.2 mmol) was added and the reaction mixture was stirred overnight. The mixture was concentrated and the residue was diluted with water (100 ml) and acidified with 3 N HCl solution to pH = 3. The resulting precipitate was collected by filtration to give the title compound (1.06 g, 1.56 mmol, 86% yield) as a yellow-white solid. ¹ H NMR (400 MHz, DMSO ^ ₆ ) δ ppm 12.80-12.87 (m, 2H), 7.93-7.98 (m, 3H), 7.85 (d, J = 8.3 Hz, 1H) 7.71 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H ), 7.33 (s, 1H), 6.54 (d, J = 6.6 Hz, 2H), 5.90-5.98 (m, 2H), 4.86 (d, J = 20, 0 Hz, 4H), 4.52 (d, J = 6.7 Hz, 4H), 2.12 (s, 6H), 1.27 (m, J = 6.8, 4.6 Hz, 6H) ; LCMS (LCMS Method A): retention time = 1.376 min, [M + H] + = 677.9.

Example 84.23- (Aminomethyl) -8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,

28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'1 ] [1,3,6,15,17] pentaazacyclogenaicosin-3-carboxylic acid

Example 84 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of obtaining is given: to tert-butyl ((3-carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21 , 28,29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3' -1] [1,3,6,15,17] pentaazacyclogenaicosin-23yl) methyl) carbamate (410 mg, 0.509 mmol) was slowly added TFA (3 ml, 38.9 mmol). After 30 minutes, the reaction mixture was concentrated. The resulting residue was suspended in water and treated with NaHCO ₃ to basic pH. The solid was collected by filtration and washed with water. This material was purified on silica gel (80 g column) eluting with 0-20% (2N NH ₃ in MeOH) in DCM to give the title compound (340 mg, 0.458 mmol, 90% yield) as an off-grade white solid. 1H NMR (400 MHz, DMSO-dg) δ ppm 8.02 (d, J = 1.2 Hz, 1H), 7.98 (brs, 1H), 7.82 (dd, J = 8.3 , 1.5 Hz, 1H) 7.60 (d, J = 8.3 Hz, 1H), 7.48 (d, J = 8.1 Hz, 1H), 7.34 (br s, 1H) , 7.24 (t, J = 7.8 Hz, 1H) 7.14 (d, J = 7.6 Hz, 1H), 6.55 (s, 1H), 4.74 (t, J = 7 , 1 Hz, 2H), 4.47 (q, J = 7.0 Hz, 2H), 4.18-4.32 (m, 4H), 4.12 (s, 2H), 2.77-2 , 88 (m, 2H), 2.15 (s, 3H), 2.05-2.11 (m, 3H), 1.91 (br s, 4H), 1.77-1.86 (m , 2H), 1.431.56 (m, 2H), 1.38 (m, 2H), 1.30 (t, J = 7.1 Hz, 3H); LCMS (LCMS Method D): retention time = 0.7 7 min, [M + H] ⁺ = 705.6.

Example 85 (E) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1 - (4- (2- (1-ethyl-3-methyl-1 H-pyrazole 5 -carboxamido ) -4-methyl-1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -1H-benzo Schimidazole-5-carboxamide

- 138 036435

Example 85 can be prepared according to method 11 with modifications known to one of ordinary skill in the art. The last stage of the preparation is given: HATU (616 mg, 1.62 mmol) and DIEA (1-ethyl-3-methyl-1Н-pyrazole-5-carboxylic acid (208 mg, 1.35 mmol) were added to NMP (8 ml). 0.57 ml, 3.2 mmol). After 30 min, (E) -2-amino-1- (4- (2-amino-4-methyl-1Nbenzo-imidazol-1-yl) but-2-en-1-yl) -1H-benzo-imidazole-5-carboxamide dihydrobromide (290 mg, 0.540 mmol) and the reaction mixture was heated to 60 ° C. After stirring overnight, water was added and the mixture was extracted with EtOAc. The organic layer was concentrated and purified by preparative HPLC to give the title compound (45 mg, 0.069 mmol, 13% yield) as an off-white solid. ¹ H NMR (400 MHz. DMS.'OD ₆ ) δ ppm 12.79 (br s, 1H), 12.19 (br s, 1H), 7.97 (s, 1H), 7.96 ( s, 1H), 7.72-7.75 (m, 1H), 7.44-7.47 (m, 1H), 7.37 (s, 1H), 7.24-7.27 (m , 1H), 7.00-7.12 (m, 2H), 6.56 (s, 2H), 5.78-5.89 (m, 2H), 4.75-4.85 (m, 4H ), 4.49-4.56 (m, 4H), 2.48-2.52 (m, 3H), 2.13 (s, 6H), 1.211.29 (m, 6H); LCMS (LCMS Method A): retention time = 1.507 min, [M + H] + = 648.2.

Example 86 (E) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1- (4- (2- (1-ethyl-3-methyl-1H-pyrazole 5 -carboxamido) -7-methoxy-1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -1H-benzo [d] imidazole-5 carboxamide

Example 86 can be prepared according to method 9 with modifications known to one of ordinary skill in the art. The last stage of preparation is presented: to 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (295 mg, 1.92 mmol) in NMP (8 ml), HATU (874 mg, 2.30 mmol) was added at room temperature and DIEA (0.803 ml, 4.60 mmol). After 30 min (E) -2-amino-1- (4- (2amino-7-methoxy-1H-benzo [d] imidazol-1-yl) but-2-en-1-yl) -1H-benzo [d] imidazole-5-carboxamide (300 mg, 0.766 mmol) and the reaction mixture was heated to 60 ° C overnight. Water was added and the resulting solid was collected by filtration and purified by preparative HPLC to give the title compound (78 mg, 0.12 mmol, 15% yield). 1H NMR (400 MHz, DMSOD ₆ ) δ ppm 12.75 (s, 2H), 7.95 (d, J = 12.1 Hz, 2H), 7.70 (d, J = 8.4 Hz, 1H ), 7.40 (d, J = 8.4 Hz, 1H), 7.33 (s, 1H), 7.17-7.09 (m, 2H), 6.80 (dd, J = 6, 8, 2.4 Hz, 1H), 6.53 (d, J = 17.1 Hz, 2H), 5.92-5.96 (m, 1H) 5.71 (m, 1H), 4.91 (d, J = 4.9 Hz, 2H), 4.81 (d, J = 5.4 Hz, 2H), 4.52 (q, J = 7.1 Hz, 4H), 3.69 (s , 3H), 2.11 (d, J = 4.0 Hz, 6H), 1.26 (td, J = 7.1, 2.0 Hz, 6H); LCMS (LC-MS Method A): retention time = 1.494 min, [M + H] + = 664.2.

Example 87.4 - ((Dimethylamino) methyl) -8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28, 29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacyclogeneicosin-2 4-carboxamide

- 139 036435

Example 87 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: a mixture of 4- (aminomethyl) -8ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28,29, 30.31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1 , 3,6,15,17] pentaazacyclogeneicosine-24carboxamide (100 mg, 0.142 mmol) and formaldehyde (37% in water) (0.106 ml, 1.42 mmol) in acetic acid (5 ml) was stirred at room temperature for 20 min and then cooled to 0 ° C. Sodium triacetoxyborohydride (301 mg, 1.42 mmol) was added, the reaction mixture was stirred at 0 ° C for 1 h and then at room temperature overnight. The solvent was removed and the residue was suspended in water and treated with NaHCO ₃ until basic pH was obtained. The mixture was extracted with EtOAc (3x) and the combined organic layers were washed with brine (2x), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified on silica gel (40 g column) eluting with 0-20% MeOH in DCM to give the title compound (30 mg, 0.039 mmol, 27% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSO-dg) δ ppm 12.89 (brs, 1H), 12.24 (1H), 7.96 (d, J = 8.31 Hz, 1H), 7.51 (d, J = 8.44, 1.59 Hz, 1H), J = 7.82 Hz, 1H), 7.35 (br s, 1H), 7.26 (t, J = 7.83 Hz , 1H), 7.14 (d, J = 7.58 Hz, 1H), 1H), 4.74 (t, J = 7.09 Hz, 2H), 4.47 (q, J = 7, 01 Hz, 2H), 4.13-4.34 (m, 4H), 3.73 (s, 2H) - 2.88 (m, 2H), 2.23 (s, 6H), 2.16 ( s, 3H), 2.09 (s, 3H), 1.92 (brs, 4H), 1.81 (m, 2H), 1.49 (m, 2H), 1.39 (m, 2H ), 1.29 (t, J = 7.09 Hz, 3H); LCMS (LCMS Method D): retention time = 0.80 min, [M + H] + = 733.6.

Example 88 tert-Butyl ((3-carbamoyl-8-ethyl-10,18-dimethyl-7,20-dioxo-6,7,8,11,12,13,14,15,20,21,28, 29,30,31-tetradecahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'-1] [1,3,6,15,17] pentaazacycloneicosin-2 3-yl) methyl) carbamate

Example 88 can be prepared according to method 13 with modifications known to one of ordinary skill in the art. The last stage of preparation is given: to 4- (5- (5 - ((1- (4- (2-amino5-carbamoyl-1H-benzoSHimidazol-1-yl) butyl) -4 - (((tert-butoxycarbonyl) amino) methyl) -1H-benzoH-imidazol-2-yl) carbamoyl) -3-methyl-1H-pyrazol-1-yl) pentyl) -1-ethyl-3-methyl-1H-pyrazole-5carboxylic acid (1.50 g, 1 , 82 mmol), HOBt (0.335 g, 2.19 mmol) and EDC (0.52 4 g, 2.7 3 mmol) in DMF (100 ml) was added DIEA (1.27 ml, 7.29 mmol) and DMAP (22 mg, 0.18 mmol). The reaction mixture was heated to 60 ° C. After 6 h, the mixture was cooled to room temperature, diluted with water and extracted with EtOAc (3x). The combined organic fractions were washed with saturated NH ₄ Cl solution (2x) and brine (2x), dried over Na ₂ SO ₄ , filtered and concentrated. This residue was purified on silica gel (120 g column) eluting with 0-20% MeOH in DCM to give the title compound (860 mg, 1.0 mmol, 56% yield) as an off-white solid. 1H NMR (400 MHz, DMSOO δ ppm 12.85 (s, 1H), 12.41 (s, 1H), 8.02 (d, J = 1.5 Hz, 1H), 7.98

- 140 036435 (br s, 1H), 7.82 (dd, J = 8.6, 1.5 Hz, 1H), 7.60 (d, J = 8.6 Hz, 1H), 7.57 (br s, 1H), 7.54 (d, J = 8.1 Hz, 1H), 7.34 (br s, 1H), 7.27 (t, J = 7.8 Hz, 1H) , 7.15 (d, J = 7.6 Hz, 1H), 6.55 (s, 1H), 4.74 (t, 7.1 Hz, 2H), 4.48 (q, J = 7, one

Hz, 2H), 4.37 (d, J = 5.9 Hz, 2H), 4.25 (m, 4H), 2.75-2.87 (m, 2H), 1.15 (s, 3H ), 1.19 (m, 11H), 1.30 (t, J = 7.1 Hz,

3H); LCMS (LCMS Method D): retention time = 1.11 min, [M + H] + = 805.7.

Table 1 lists examples 89-197. Compounds can be prepared using the synthetic methods described above.

Example Structure / Name Synthesis method NMR LC-MS Example 89 1 - (((4R, 5R) -5 - ((5carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-1yl) methyl) -2,2-dimethyl1, 3 -dioxolan-4-yl) methyl) -2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -7- (3hydroxypropoxy) -1Hbenzo [d] imidazole-5carboxamide OH nh ₂ \ h ₂ n V \ C? \ _J NH HN O ^ Z ι NsS ° < ^z 1 1 7 CsN Ni; / Method 9 ² H NMR (4 00 MHz, (methanol-b ₄ δ 7.58 (s, 1H), 7.48 (d, J = 8.4 Hz, 1H), 7.33 (s, 1H), 7, 17 (d, J = 8.4 Hz, 1H), 6.89 (s, 1H), 6.58 (s, 2H), 5.11-4.99 (m, 1H), 4.93 (s , 1H), 4.61 (m, 5H), 4.4 6-4.36 (m, 1H), 4.31 (dd, J = 13.4, 3.2 Hz, 1H), 4.19 (dd, J = 14.0, 3.4 Hz, 1H), 4.03 (dd, J = 15, l, 6.4 Hz, 1H), 3.88 (dd, J = 14.9, 6 , 4 Hz, 1H), 3.83-3.72 (m, 1H), 2.23 (s, 3H), 2.18 (s, 3H), 2.05 LC-MS Method A Retention Time = 1.33 8 min, [M + H] ⁺ = 8 24.8

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Example

1- (4- (4 ((dimethylamino) methyl) -2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butyl) -2- (1-ethyl- Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide trifluoroacetate

Example

(dd, J = ll, 5.5, 5.9 Hz, 2H), 1.64 (d, J = 10.3 Hz, 6H), 1.50- 1.28 (m, 6H) Way eleven ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.86 (s, 2H), 9.55 (s, 1H), 7.99 (s, 2H), 7.78 (s, 1H), 7.65 (dd, J = 6.7, 2.2 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.33 (t, J = 7.9 Hz, 3H ), 6.61 (d, J = 9.9 Hz, 2H), 4.68 (d, J = 4.9 Hz, 2H), 4.56 (dd, J = 13.9, 6.8 Hz , 4H), 4.28 (s, 4H), 2.77 (d, J = 4.5 Hz, 6H), 2.11 (s, 6H), 1.87 (s, 4H), 1.30 (t, J = 7.0 Hz, 6H) LC-MS Method A retention time = 1.27 5 min, [M + H] ⁺ = 6 93.3 Method 2 ^X H NMR LC-MS

(E) -1,1 '- (but-2-ene-1,4-

diyl) bis (2- (1-ethyl-3- (methanol-b ₄ ) δ method F: methyl-1H-pyrazole-5- 7.52-7.65 (m, time carboxamido) -7- (3- 2H), 7.23- holding methoxypropoxy) -1H- 7.37 (m, 2H), nia = 0.87 benzo [d] imidazole-5- 6, 55-6, 69 (m, min, carboxamide) 2 H), 5.78- [M + H] ⁺ 5.93 (m, 2H), = 853.7

5.03-5.10 (m,

- 142 036435

) 5 NN N __n Jn— \ I II O / T ^h o '\ / ίΑγ ^Ν ' Il I Z> - ^NH , ⁿ -n ^Η 2 ^Ν χΛΑν 4H), 4.49- 4.70 (m, 4H), 3.98-4.10 (m, 4H), 3.36- 3.46 (m, 6H), 2.24 (s, 6H), 1.71-1.90 (m, 4H), 1.35- 1.47 (m, 6 N) Example 92 1,1 '- (cyclopentane-l, 3diyl) bis (2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -7- (3hydroxypropoxy) -1Hbenzo [d] imidazole-5carboxamide)) p NN and _m 11 if ^NH2 \ Η X -Ν but LJ ^ n / \\ Lu / 1 Α ~ ^Ν ο η ₂ ν ^ / 0 Way 15 ^X H NMR (4 00 MHz, methanol d ₆ ) δ ppm 7.59-7.78 (m, 2Η), 7.46-7.55 (μ, 2Η), 6.57-6.72 (μ, 2 ), 5.78- 6.04 (μ, 2Η), 4.48-4.71 (μ, 4Η), 4.32- 4.45 (μ, 4Η), 3.74-3.89 (μ , 4Η), 2.92- 3.02 (μ, 2Η), 2.29-2.44 (μ, 2Η), 2.15- 2.26 (μ, 4Η), 2.09 (s, 6Η ), 1.32 (μ, 6Η), 0.84-0.99 (μ, 2Η) LC-MS Method F Retention Time = 0.8 0 min, [M + H] ⁺ = 8 39.7 Example 93 ethyl (Ε) -3- (5-carbamoyl1- (4- (5-carbamoyl-2- (1ethyl-3-methyl-1H-pyrazole5-carboxamido) -1Nbenzo [d] imidazole-1yl) but-2-ene- 1-yl) -2- (1ethyl-3-methyl-1H-pyrazole 5-carboxamido) -1Hbenzo [d] imidazole-7yl) propanoate Way ten ^Χ Η NMR (4 00 MHz, DMSO-dg) δ ppm 12.6613.00 (μ, 2Η), 7.89-8.05 (m, 2Η), 7.88 (s, 1Η), 7.71 (d, J = 8, ll Hz, 1Η), 7.53 s, 1 Η), 7.42 LC-MS Method F Retention Time = 0.8 6 min, [M + H] ⁺ = 777.5

- 143 036435

Example

(d, 0 = 8.36

Hz, 1H), 7.34 (d, 0 = 9.38

Hz, 2H), 6.53 (D, 0 = 3.55

Hz, 2H),

5.95-6.07 (m,

1H), 5.495.65 (m, 1H), 4.96-5.08 (m,

2H), 4.794.88 (m, 2H), 4.37-4.63 (m,

4H), 3.97 (q, 0 = 7.10

Hz, 2H),

3.03-3.15 (m,

2H), 2.63 (t, 0 = 7.73 Hz, 3H), 2.12 (d, 0 = 6.08 Hz, 6

H), 1.28 (d,

0 = 3.80 Hz, 6

H), 1.09 (t,

0 = 7.10 Hz, 3

H) methyl (E) -4- ((5 - carbamoyl-1- (4- (5carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-1yl) but-2 -en-1-yl) -2- (1ethyl-3-methyl-1H-pyrazole 5-carboxamido) -1Hbenzo [d] imidazole-7yl) oxy) butanoate

Way

^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.7112.97 (m, 2H), 7.98 (d, 0 = 1.27 Hz, 2H), 7.91-7.96 (m , 1H), 7.67-7.76 (m, 1H), 7.59- 7.66 (m, 1H), 7.42 (d, 0 = 7.60 Hz, 1H), 7.33- 7.39 (m, 2H), 7.28-7.32 (m, LC-MS Method K: retention time = 0.85 min, [M + H] ⁺ = 7 93.5

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^h o ° ^ 1 nh ₂ L Q y = N T HN% W ^H \ =: O G Ν—. \ = N Learn \ 1H), 6.47- 6.58 (m, 2H), 5.92-6.05 (m, 1H), 5.67- 5.80 (m, 1H), 4.91-4.99 (m, 2H), 4.78- 4.87 (m, 2H), 4.46-4.58 (m, 4H), 4.03 (s, 2H), 3.55 (s, 3H), 2.29- 2.39 (m, 2H), 2.12 (d, J = 3.80 Hz, 6H), 1.71- 1.93 (m, 2H), 1.22-1.32 (m, 6H) Example 95 (E) -1- ((E) -4- ((E) -5carbamoyl-2 - ((1-ethyl-Zmethyl-1H-pyrazole-5carbonyl) imino) -2,3dihydro-1H- benzo [d] imidazole-1- yl) but-2-en-1-yl) -2 - ((1ethyl-3-methyl-1H-pyrazole5-carbonyl) imino) -7- (3hydroxy-2- methylpropoxy) -2,3dihydro-1H- benzo [d] imidazole-5carboxamide Way 14 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.6212.93 (m, 1H), 7.91-8.05 (m, ZN), 7.71 (d, J = 8, ll Hz , 1Η), 7.58- 7.68 (μ, IH), 7.41 (d, J = l, 86 Hz, 1Η), 7.28- 7.37 (μ, 3Η), 6.47- 6.55 (μ, 2Η), 5.93- 6.08 (μ, 1Η), 5.70-5.84 (μ, 1Η), 4.90- 5.00 (m, 2Η), 4, 77-4.87 (μ, 2Η), 4.62 (τ, J = 5.80 Hz, LC-MS Method K: retention time = 0.81 min, [M + H] ⁺ = 7 65.5

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6 N) Example 97 (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 7- (3-methoxypropoxy) -1Hbenzo [d] imidazole-1yl) but-2 -en-1-yl) -2- (1ethyl-3-methyl-1H-pyrazole5-carboxamido) -7isobutyl-1H-benzo [d] imidazole-5carboxamide trifluoroacetate /? d '^ ^Ν ^ Γ ^ίίί Αι ^4ΝΗ2 Λ-у ην - ς' 1 | l ° χ ϊ AA ' ^N Method 9 ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.91 (br s, 1H), 7.89-8.16 (m, 4H), 7.64 (s, 1H), 7.47 (br s, 1 H) 7.25-7.40 (m, 2H) 6.47-6.60 (m, 2H) 5.82 (d, J = 15.72 Hz, 1H) 5.54 (d, J = 15.72 Hz, 1 H), 4.91 (br s, 4H) 4.41-4.65 (m, 4H), 4.03 (t, J = 6.46 Hz, 2 H) 3.29 (t, J = 6.0 8 Hz, 2H), 3.14 (s, 3H), 2.55 (br s, 2H), 2.11 (d, J = 12, 17 Hz, 6H) 1.69-1.84 (m, 2H), 1.36 (m, 1H), 1.27 (q, J = 7.10 Hz, 6H), 0.69 (s, J = 6.3 Hz, 6H) LC-MS Method F Retention Time = 2.33 min, [M + H] ⁺ = 8 22.1 Example 98 (Ε) -1- (4- (5-carbamoyl-2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) IH-benzo [d] imidazol-1yl) but-2-en-1-yl) -2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7 (methoxymethyl) -1H- Method 9 ^X H NMR (4 00 MHz, DMSO- ₆ ) δ ppm 12.98 (s, 1H), 7.98 (m, 4H), 7.77-7.65 (m, 2H), 7.44 ( d, J = 8.4 Hz, LC-MS Method A Retention Time = 1.30 min, [M + H] ⁺ = 7 21.4

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benzo [d] imidazole-5carboxamide .O jj ^N \\ X ζ, ^Ννη 2 ην — ς 1 | Ι ϊ Α- 2H), 7.35 (s, 2H), 6.53 (d, J = 6.8 Hz, 2H), 5.95 (d, J = 15.8 Hz, 1H), 5.52 (d, J = 15.8 Hz, 1H), 4.95 (s, 2H), 4.81 (s, 2H), 4.52 (d, J = 1.0 Hz, 6H), 3.15 (s, 3H), 2.11 (s, 6H), 1.26 (td, J = 1.1, 3.7 Hz, 6H) Example 99 (Ε) -2- (5-carbamoyl-1- (4- (5-carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Nbenzo [d] imidazole-1yl) but-2-ene -1-yl) -2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [d] imidazole-7yl) acetic acid /? n N \ ___ // .. 11 \\ # \ HN — C I | 1 \ fl OH G | G NH _g __ / P A XN O ° _ ^ Method 9 ^X H NMR (4000 MHz, flMCO-d ₆ ) δ ppm 12.87 (s, 3H), 8.02-7.87 (m, 4H), 7.71 (d, J = 8.4 Hz, 1H), 7.57 (s, 1H), 7.41 (d, J = 8.4 Hz, 1H), 7.32 (s, 2H), 6.52 (d, J = 14.4 Hz, 2H), 5.95 (d, J = 15.0 Hz, 1H), 5.48 (d, J = 15.5 Hz, 1H), 4.97 (s, 2H), 4.79 (s, 2H), 4.59- 4.46 (m, 4H), 3.71 (s, 2H), 2.08 (t, J = 12, l Hz, 6H), 1.25 (td, J = 1 ,one, LC-MS Method A Retention Time = 1.30 min, [M + H] ⁺ = 7 35.4

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Example

one hundred

1- (4- (5-carbamoyl-2- (1- Method ethyl-3-methyl-1H-pyrazole-16

5-carboxamido) -1Нbenzo [d] imidazol-1-yl) 2,3-dimethylbutyl) -2- (1ethyl-3-methyl-1Н-pyrazole-5-carboxamido) -7- (3hydroxypropoxy) -1Нbenzo [d] imidazole-5carboxamide

2.6 Hz, 6H) ^X H NMR (400 Time MHz, DMSO-dg) holding 8 ert 7.76 niya A: (d, J = 9.6 Hz, time 1H), 7.55- holding 7.47 (m, 2H), nia = 1.33 7.14 (s, 1H), 5 & 7.03 (d, 1,362 J = 9, 7 Hz, MIN, 2H), 6.62 (s, [M] ⁺ = 780 1H), 6.56 (s, 1H), 4.62- 4.50 (m, 4H), 4.35-4.17 (m, ZN), 4.05- 3.97 (m, 1H), 3.78-3.70 (m, 2H), 3.38- 3.32 (m, 2H), 2.68 (s, 1H), 2.34 (s, 1H), 2.16 (s, 3H), 2.14 (s, 3H), 2.08-1 , 96 (m, 2H), 1, δδ- Ι, 48 (m, 2H), 1.38-1.28 (m, 4H), 1.22 (s, 2H), 1.04 (d, J = 6.7 Hz, ZN), 0.97 (d, J = 6.7 Hz, ZN), 0.91 - 0.80 (m, 2H) (NMR analysis based on main peaks , 7

main diastereomer

Example (E) -7- (2-aminoethyl) -1- (4 Analogies

D NMR (4 00

LC-MS

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0 Ι \ Κ _Ν ΗΝ— ('Ι | 1 / ΝΗ ⁰ 0 (q, J = 7.1 Hz, 2Η), 4.34 (τ, J = 6.9 Hz, 2Η), 3.76 (τ, J = 6, 7 Hz, 2Η), 3.68- 3.63 (m, 2Η), 2.84-2.78 (μ, 2Η), 2.24 (s, ZN), 2.18 (s, ZN), 1.89- 1.82 (m, 2Η), 1.42 (dd, ι7 = 13.9, 6.4 Hz, 2Η), 1.34 (τ, J = 7.1 Hz, ZN), 1.27 - 1.19 (m, 2Η) Example 103 8-ethyl-10, 18-dimethyl-7,20-dioxo-7,8,11,12,13, - 14,15,20,21,28,29,30,31,3 2-tetrahydro-6Η- benzo [4,5] imidazo [2,1b] benzo [4,5] imidazo [1,2-i] dipyrazolo [5,1-m: 4 ', 3' t] [1,3,6,9, 11,14] hexaazacyclodocosine-24carboxamide o 9 ^{N H} 2 N ^ n ΗΝ — y Π 1 \ Ν · ^ ⁴ ^ / NH \ __ / _n N ^ x ^ 4, W ^ ^N— ΊΓ Η Ν Π ^Ν ο Way 13 ^Χ Η NMR (400 MHz, (methanol-cC) δ 8.08-8.06 (μ, 1Η), 7.94-7.90 (μ, 1Η), 7.63 -7.59 (μ, 2Η ), 7.55-7.51 (μ, 1Η), 7.42- 7.34 (μ, 2Η), 6.85-6.82 (μ, 1Η), 4.75- 4.69 (μ , 4Η), 4.53-4.46 (μ, 3Η), 4.36- 4.30 (μ, 2Η), 3.82-3.77 (μ, 2Η), 3.69- 3.65 (m, 2Η), 2.83-2.76 (μ, 2Η), 2.25- 2.21 (μ, 3Η), 2.19-2.17 (μ, LC-MS Method A Retention Time = 1.2 9 7 min, [M + H] ⁺ = 6 91.3

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Example

104

8-ethyl-10, 18,30-trimethyl7,20-dioxo7,8,11,12,13,14,15,20,21,28,29,30,31,3 2-tetrahydro-6Hbenzo [4.5 ] imidazo [2,1 -

B] benzo [4,5] imidazo [1,2 - i] dipyrazolo [5,1-t: 4 ', 3't] [1,3,6,9,11,14] hexaase cyclodocosine-3carboxamide

Way

05.3

ZN), 1.87 (s, 2H), 1.46- 1.41 (m, 2H), 1.33- 1.32 (m, ZN), 1.25- 1.19 (m, 2H)

(400 LC-MS method A: cs) δ time 1H), holding (d, nia = 1.32 HZ, MIN, 7.56 [M + H] ⁺ = 7

7.39-7.35 (m, 2H), 6.75 (FROM, 1H), 4.78 (FROM, 2H), 4.68 (FROM, 2H), 4.53

(dd, <J = = 14.3, 7.2 Hz, 4H), 4,00 (FROM, 2H), 3.78 (from, 2H), 3.15 (from, ZN), 2.83 (from, ZN), 2.35 (d, J = 7, 8 Hz, 2H), 2.17 (s, 6H), 1.7 7 (c, 2H), 1.36 (dd, <J = = 15.7, 8.6 Hz, 6H), 1.14 (FROM, 2H)

Example 35-ethyl-5,37-dimethyl- Way 105 8.33-dioxo3.4.9.11.18.23.30.32.35.36-decaazoctacyclo [38.3.1.0 3 7 Q 1 0 I ⁸ Q ^{12 17} O ^{23 31} 0 g ⁴ ₂ θ, o ^s4 , ^s8 ] ^{tetratetracon} ta-1 (44), 4,6,10, - 19

^X H NMR (4 00 LC-MS MHz, DMSO-s1 ₆ ) method δ ppm 12.86 A: time (br. s, 2H) holding 8.00-8.01 (m, nia = 1.38 4H) 7.77-7.81 6 min, (m, 2H) 7.52- [M + H] ⁺ = 7 7.58 (m, 2H) 53.2

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methyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide) di-trifluoroacetate ABOUT II 0 y—- A ABOUT \ δ ppm 12.85 (br s, 2H) 7.99 (s, 4H) 7.77 (d, 3 = 8.34 Hz, 2H), 7.56 (d, 3 = 8.34 Hz, 2H), 7.36 (br s, 2H), 6.61 (s, 2 H), 4.51-4.69 (m, 4H) 4.28 (br s, 4H), 2.04-2.21 (m, 6H), 1.88 (br s, 4H), 1.31 (t, 3 = 7.07 Hz, 6H) retention time = 0.7 6 min, [M + H] ⁺ = 6 79.6 Example 108 1- (4- (5-carbamoyl-2- (1,3- dimethyl-1H-pyrazole-5carboxamido) -1Nbenzo [d] imidazol-1- yl) butyl) -2- (1-ethyl-Zmethyl-1H- pyrazole-5carboxamido) -1Нbenzo [d] imidazole-5carboxamide ditrifluoroacetate ^HiN iX ι> Ar o \ Way twenty ^X H NMR (4 00 MHz, flMCO-d ₆ ) δ ppm 12.82 (br s, 2H), 7.98 (s, 4H), 7.76 (dd, 3 = 8.31, 1.47 Hz, 2H), 7.55 (D, 3 = 8.31 Hz, 2H), 7.35 (br s, 2H), 6.60 (s, 2H), 4.57 (q, 3 = 7 , 01 Hz, 2H) 4.28 (d, 3 = 5.87 Hz, 4H), 4.09 (s, 3 H), 2.10 (D, 3 = 4.65 Hz, 6H), 1, 87 (br s, 4H), 1.30 (t, 3 = 7.09 Hz, LC-MS Method D Retention Time = 0.77 min, [M + H] ⁺ = 6 65.6

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ZN) Example 109 1- (4- (5-carbamoyl-2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [d] imidazole-1yl) butyl) -2- (3cyclopropyl-1-methyl-1Hpyrazole- 5-carboxamido) 1H-benzo [d] imidazole-5carboxamide ditrifluoroacetate X ii 1 ^Η2Ν Λ) O \ Way twenty ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.84 (br s, 2H), 7.99 (s, 4H), 7.76 (dd, J = 8.56, 1.47 Hz , 2H), 7.54 (d, J = 8.31 Hz, 2H) 7.35 (br s, 2H) 6.66 (s, 1H) 6.60 (s, 1H) 4.57 (q , J = 7.01 Hz, 2H) 4.26-4.39 (m, 4H) 4.09 (s, 3H) 2.09 (s, 3H) 1.701.90 (m, 5H) 1.31 ( t, J = 7.09 Hz, 3H) 0.72-0.83 (m, 2H) 0.460.57 (m, 2H) LC-MS Method D Retention Time = 0.84 min, [M + H] ⁺ = 6 91.6 Example 110 (11Z, 2 9E) -8-ETHYL-1,2 6bis (3-hydroxypropoxy) 10,18-dimethyl-7,20dioxo 6,7,8,13,14,15,20,21,28,3 1- decahydrobenzo [4,5] imidaz o [1,2- a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1e: 4 ', 3'1] [1,3,6,15,17] pentaazacycloneicosin- 3,24-dicarboxamide dihydrochloride Way 19 ^X H NMR (4 00 MHz, (methanol-b ₄ ) δ δ ppm 7.71 (dd, J = 5.32, 1.01 Hz, 2H), 7.44 (dd, J = 8.74, 1 , 14 Hz, 2H), 6.82 (s, 1H), 6.46 (d, J = ll, 15 Hz, 1H), 5.84-5.96 (m, 1H), 5.58 (s , 2H), 5.09 (br. S,, 4 LC-MS Method L retention time = 0.7 5 min, [M + H] ⁺ = 8 63.7

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О H ₂ nX S '/ / - ^N \\ N \\ Άί ^Ν HO, Y == X S ^ N \\ // \ / Ν Η Y— \ Α ϊ no <XG''\ * A— NH N_ ... n ^ YES ^G VAji J o H), 4.62-4.75 (m, 2H), 4.07-4.23 (m, 4H), 3.72- 3.79 (m, 4H), 3.53-3.58 (m, 2H), 2.33 (s, 3H), 2.26 (s, 3H), 1.93- 2.11 (m, 4H), 1.54-1.71 (m, 4H), 1.49 (t, J = 7.10 Hz, ZN); Example 111 (E) -1,1 '- (but-2-ene-1,4diyl) bis (2- (1-ETHYL-4fluoro-3-methyl-1H-pyrazole5-carboxamido) -7- (3hydroxypropoxy) -1Нbenzo [ d] imidazole-5carboxamide) F xj О ° J '// ^{Ν n} 2 Ν ΗΝ — С) \ ° χ / Χ> Ή aJ <О ° F Method 2 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.90 (s, 2H), 7.99 (br.s, 2H), 7.64 (d, J = 1, 27 Hz, 2H), 7.29-7.38 (m, 4H), 5.87 (br s, 2 H), 4.91 (br s, 4H), 4.40-4.62 (m, 6 H), 4.04 (t, J = 6.08 Hz, 4H), 3.43 (d, J = 5.07 Hz, 4H), 2.10 (s, 6H), 1.67 (t, J = 6 , 08 Hz, 4H), 1.24 (t, J = 7.10 Hz, 6H); LC-MS Method D Retention Time = 0.8 9 min, [M + 2] ⁺ / 2 = 431.5 Example 112 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) - 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5- carboxamido) -4- Way 16 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.80 (s, 1H), 12.57 (s, LC-MS method A: retention time = 1.27

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(morpholinomethyl) -1Hbenzo [d] imidazol-1yl) butyl) -1H-benzo [d] imidazole-5carboxamide / ° H ⁿ n hn —c; JU (B JL / N τ ^ ^Ν JL 1ST 1H), 7.98 (s, 2H), 7.77 (d, 0 = 8.2 Hz, 1H), 7.54 (d, 0 = 8.4 Hz, 1H), 7.30- 7.48 (m, 2H), 7.14 (d, 0 = 27.8 Hz, 2H), 6.59 (s, 2H), 4.48- 4.67 (m, 4H), 4.27 (s, 4H), 3.81 (s, 2H), 3.70 (s, 4H), 2.42 (s, 4H), 2.11 (s , 6H), 1.88 (s, 4H), 1.30 (td, 0 = 7.0, 3.4 Hz, 6H) 5 min, [M + H] ⁺ = 7 35.3 Example 113 tert-butyl ((1- (4- (5carbamoyl-2- (1-ethyl-Zmethyl-1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-1yl) butyl) -2- (1-ethyl-Zmethyl-1Н -pyrazole-5carboxamido) -1Нbenzo [d] imidazole-4yl) methyl) - (methyl) carbamate ^N N HN — I P) (B JL /> ~ \ ⁿ / 'n о УЧ ίι Lx JI 1 О' • 'X xXg'yy 1 Way 16 ^X H NMR (400 MHz, flMCO-d ₆ ) δ ppm 12.81 (s, 2H), 7.99 (t, 0 = 5.1 Hz, 2H), 7.78 (d, 0 = 8, 4 Hz, 1H), 7.55 (d, 0 = 8.4 Hz, 2H), 7.48 (d, 0 = 8.2 Hz, 1H), 7.34 (s, 1H), 7.21 (d, 0 = 7.7 Hz, 2H), 6.60 (d, 0 = 12.2 Hz, 2H), 4.48-4.77 (m, 6H), 4.27 (d, 0 = 5.9 Hz, 4H), 2.72 (s, LC-MS Method A Retention Time = 1.64 6 min, [M + H] ⁺ = 7 79.3

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ZN), 2.09 (t, J = 4.3 Hz, 6H), 1.87 (s, 4H), 1.44 (s, 9H), 1.28 (td, J = 7.0, 2.4 Hz, 6H) Example 114 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) - 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4isopropyl-1H-benzo [d] imidazole- 1-yl) butyl) -1Н-benzo [d] imidazole-5carboxamide О ° nI / // ^ΝΗ ? ^N N HN - e) Way 16 ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.83 (s, 1H), 12.21 (s, 1H), 7.98 (d, J = 8.2 Hz, 2H), 7, 77 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.30- 7.41 (m, 2H), 7.07-7.28 (m, 2H), 6.60 (d, J = 10.0 Hz, 2H), 4.57 (d, J = 6.8 Hz, 4H), 4.28 (s, 4H), 3.40 - 3.54 (m, 1H), 2.11 (s, 6H), 1.87 (s, 4H), 1.05-1.47 (m, 12H) LC-MS Method A retention time = 1.60 1 min, [M + H] ⁺ = 6 78.5 Example 115 1,1 '- ((2R, 3R) -2,3dimethoxybutane-1,4diyl) bis (2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide) Way 15 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.99 (br s, 2H) 8.01 (s, 4H), 7.81 (d, J = 8.56 Hz, 2H), 7 , 55 (d, J = 8.80 Hz, 2 H), 7.36 LC-MS Method A retention time = 1.2 9 min, [M + H] ⁺ = 7 39.8

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nh ₂ h ₂ n Έ u & N ^ N— * '' —N ^ N O ^ NH HN_O la A ^J N = / \ = N (br s, 2Η), 6.64 (s, 2H), 4.53-4.70 (m, 6H), 4.46 (dd, J = 14.06, 8.68 Hz, 2H), 4.04 (br s, 2H), 3.21 (s, 6H), 2.11 (s, 6H), 1.35 (t, J = 7.09 Hz, 6H), Example 116 (E) -1- (4- (6-carbamoyl-2 (1,3-diethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazol-1-yl) but-2-en-1-yl) - 2- (1,3diethyl-1Н-pyrazole-5carboxamido) -7- (3hydroxypropoxy) -1Нbenzo [d] imidazole-5carboxamide nh ₂ О = / у – ОН ° <S ^ - ^NH 2 Υ 7 " ^{Ν ΗΝ} χ ^ ° / ^ΗΝ \ \ I \ λ ° 1 A Ν-Ζ 7 Ο Ν = Ζ Way 16 ^X H NMR (4 00 MHz, DMSO- ₆ ) δ ppm 12.5613.10 (m, 1H), 7.99 (s, 2H), 7.94 (br s, 1H), 7.71 ( d, J = 8.56 Hz, 1H), 7.64 (s, 1H), 7.41 (d, J = 8.31 Hz, 1H), 7.34 (br s, 3H), 6, 57 (d, J = 13.20 Hz, 2H), 5.98 (s, 1H), 5.715.82 (m, 1H), 4.95 (d, J = 4.65 Hz, 2H), 4 , 84 (d, J = 4.89 Hz, 2H), 4.49-4.62 (m, 5H), 4.11 (t, J = 6.36 Hz, 2H), 3.48 (q, J = 5.71 LC-MS Method A Retention Time = 1.33 9 min, [M + H] ⁺ = 7 79.8

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Example

117

2- (1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido method) - 16

1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -7-methoxy1H-benzo [d] imidazol-1yl) butyl) -1Hbenzo [d] imidazole-5carboxamide o

Example

118

2- (1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido method) - 16

1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4-

Hz, 2 Η), 2.43-2.49 (m, 2 Η), 1.75 (t, J = 6.24 Hz, 2H), 1.28 (t, J = 7.09 Hz, 6H), 1.13 (td, J = 7.64, 3.06 Hz, 6 N) ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.82 (s, 2H), 7.98 (t, 0 = 4.5 Hz, 2H), 7.73-7.81 (m, 1H ), 7.54 (d, 0 = 8.5 Hz, 1H), 7.33 (s, 1H), 7.11-7.18 (m, 2H), 6.86 (dd, 0 = 6, 0, 3.2 Hz, 1H), 6.59 (D, 0 = 14.3 Hz, 2H), 4.57 (dt, 0 = 13.6, 6.8 Hz, 4H), 4.38 ( s, 2H), 4.28 (s, 2H), 3.81 (s, 3H), 2.11 (d, 0 = 5.9 Hz, 6H), 1.85 (s, 4H), 1, 30 (q, 0 = 7.0 Hz, 6H) LC-MS Method A retention time = 1.4 6 8 min, [M + H] ⁺ = 6 66.3 ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.84 (s, 2H), 8.67 (s, 2H), 7.98 LC-MS Method A: retention time = 1.25

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2H), 4 , 35- 4.68 (m, 6Η), 2.61- 2.82 (μ, 2H), one , 92- 2.14 (m, 6Η), 1.16- 1.39 (μ, 6H)

Example 1- (4- (4- (benzyloxy) -2- Way ^X H NMR (4 00 LC-MS 120 (1-ethyl-3-methyl-1H- eleven MHz, DMSO-dg) method D: pyrazole-5-carboxamido) - δ ppm 12.80 time 1H-benzo [d] imidazole-1- (s, 1H), holding yl) butyl) -2- (1-ethyl-3- 12.07 (s, nia = 1.18 methyl-1H-pyrazole-5- 1H), 7.98 (d, MIN, carboxamido) -1H- J = l, 5 Hz, [M + H] ⁺ = 7 benzo [d] imidazole-5- 1H), 7.94 42.5 carboxamide (br. s, 1H), 7.75 (dd, J = 8.4, 1.3 Hz, 1H), ABOUT ? N \ // 7.47-7.59 (m, 11 l— \ HN — Jj J ZN), 7.39- 7.45 (m, 2H), 7.33-7.39 (m, \] Ι Η) 1H), 7.26- Γ ¹ __ / // ΤΓ 7.32 (m, 1H), ° 7.14 (br. S, 2H), 6.92- Α ~~ νη ₂ ο ^ζ 7.02 (m, 1H), 6.59 (s, 2H), 5.30 (s, 2H), 4.43-4.62 (m, 4H), 4.15- 4.31 (m, 4H), 2.11 (s, ZN), 2.10 (s, ZN), 1.85 (br. S, 4H), 1.28 (dt, J = 14.5, 7.2 Hz, 6H) Example tert-butyl ((24- Way ^X H NMR (4 00 LC-MS 121 carbamoyl-8-ethyl-Yu, 18- 13 MHz, DMSO-b ₆ ) method D: dimethyl-7,20-dioxo δ ppm 12.90 time

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Example

122

6,7,8,11,12,13,14,15,20,2

1,28,29,30,31 tetradecabenzo [4,5] imidaz o [1,2а] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1е: 4 ', 3'1] [1,3 , 6,15,17] pentaazacycloneicosin4-yl) methyl) carbamate

BocHN

ABOUT

2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5C)

(br. s, 1H), holding 12.36 (br. nia = 1.10 s, 1H), 8.02 MIN, (br. s, 2H), [M + H] ⁺ = 8 7.84 (dd, J = 8.3, 1.3 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.62 (t, J = 6.3 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.36 (br s, 1H), 7.25 (t, J = 7.8 Hz, 1H), 7.12 (d, J = 7.8 Hz, 1H), 6.57 (s, 1H), 4.74 (t, J = 6.8 Hz, 2H), 4.47 (q, J = 7.0 Hz, 2H), 4.38 (d, J = 6.0 Hz, 2H), 4.15- 4.33 (m, 4H), 2.74-2.87 (m, 2H), 2.15 (s, 3H), 2.08 (s, 3H), 1.90 (br s, 4H), 1.80 (br s, 2H), 1.32- 1.54 (m, 2H), 1.40 (s, 9H), 1.28 (t, J = 7.0 Hz, ZN) 05.6 ^X H NMR (4 00 LC-MS MHz, DMSO-dg) method D: δ ppm 12.73 time (br. s, 1H), holding

- 163 036435

Example

123 carboxamido) -4-hydroxy1H-benzo [d] imidazol-1yl) butyl) -1Hbenzo [d] imidazole-5carboxamide

HE

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4isopropoxy-1Hbenzo [d] imidazole-1yl) butyl ) -1Нbenzo [d] imidazole-5carboxamide

Way

7.99 (d, J = l, 5 Hz, 1H), 7.81- 7.92 (m, 1H), 7.61-7.74 (m, 1H), 7.36- 7.50 (m, 1H), 7.14-7.28 (m, 1H), 6.95- 7.06 (m, 1H), 6.79-6.90 (m, 1H), 6.58- 6.64 (m, 1H), 6.49-6.58 (m, 2H), 4.49- 4.64 (m, 4H), 4.13-4.30 (m, 4H), 2.10 (s, 3H), 2.09 (s, 3H), 1.84 (br s, 4H), 1.18-1.37 (m, 6H) niya = 0.91 min, [M + H] ⁺ = 6 52.2 ^X H NMR (4 00 MHz, DMSO- ₆ ) δ ppm 12.81 (br s, 1H), 11.96 (br s, 1H), 7.98 (d, J = 1, 0 Hz, 1H), 7.96 (br s, 1 H), 7.76 (dd, J = 8.6, 1.5 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H) , 7.33 (br. S, 1 H), 7.08-7.21 (m, 2 H), 6.91 (d, J = 7, l Hz, LC-MS Method C Retention Time = 1.05 min, [M + H] ⁺ = 6 94.7

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ZN), 2.10 (s, ZN), 2.09 (s, ZN), 1.84 (lat. s, 4H), 1.29 (sq, J = 6, 8 Hz, 6H)

Example 1- (4- (4-amino-2- (1-ethyl- Way ^X H NMR (4 00 LC-MS 125 3-methyl-1H-pyrazole-5- eleven MHz, flMCO-d ₆ ) method D: carboxamido) -1H- δ ppm 12.90 time benzo [d] imidazole-1- (br s, 1H), holding yl) butyl) -2- (1-ethyl-3- 12.30 (br. nia = 0.8 8 methyl-1H-pyrazole-5- s, 1H), 7.98 MIN, carboxamido) -1H- (d, J = l, 5 Hz, [M + H] ⁺ = 6 benzo [d] imidazole-5- 1 Η), 7.96 51.6 carboxamide trifluoroacetate (br. s, 1H), ABOUT 7.76 (dd, ii y— ^Ν -ν 'ΗΝ — P Ύ jl J = 8.4, 1.6 ) ί ' ⁴ ^ Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.33 (br. s, 1H), Il T UL Ji 8 about A 6.95 (t, J = 8.0 Hz, 1H), 6.71 (d, J = 7.8 Hz, 1H), 6.61 (s, 1H), 6.56 (s, 1H), 6.49 (d, J = 8.0 Hz, 1H), 4.56 (q, J = 7.2 Hz, 4H), 4.27 (br. s, 2H), 4.14-4.23 (m,

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Example

126

Example

127

2- (1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido method) - 11

1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (Nmethylmethylsulfonamido) 1H-benzo [d] imidazole-1yl) butyl) -1Hbenzo [d] imidazole-5carboxamide trifluoroacetate

1- (4- (4- (2-aminoethyl) -2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butyl) -2- (1-ethyl- Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide trifluoroacetate

Way

2H), 2.10 (s, 3H), 2.09 (s, 3H), 1.84 (br s, 4H), 1.22-1.36 (m, 6H) ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.83 (br s, 1H), 12.05 (br s, 1H), 7.98 (s, 1H), 7.96 (br s, 1H), 7.76 (d, 3 = 8.5 Hz, 1H), 7.55 (d, 3 = 7.5 Hz, 2 H), 7.41 (br s, 1H), 7.27-7.37 (m, 2H), 6.62 (s, 2H), 4.464.62 (m, 4H), 4.27 (br s, 4H), 3.31 (s, 3H ), 3.05 (s, 3 H), 2.11 (s, 6 H), 1.86 (br s, 4H), 1.20-1.34 (m, 6H) LC-MS Method D Retention Time = 0.98 min, [M + H] ⁺ = 7 43.7 ^X H NMR (4000 MHz, DMSO-dg) δ ppm 7.98 (d, 3 = 1.3 Hz, 2H), 7.73-7.85 (m, 4H), 7.54 (d, 3 = 8.3 Hz, 1H), 7.44 (d, LC-MS Method D Retention Time = 0.7 8 min, [M + H] ⁺ = 6 79.6

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Example

128

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (Nmethylacetamido) -1Hbenzo [d] imidazole -1yl) butyl) -1Нbenzo [d] imidazole-5-

Way

0 = 7.8 Hz, 1H), 7.35 (br s, 1H), 7.17-7.24 (m, 1H), 7.09- 7.15 (m, 1H), 6.62 (s, 1H), 6.61 (s, 1H), 4.48-4.61 (m, 4H), 4.20- 4.31 (m, 4H), 3.05-3.19 (m, 4H), 2.11 (s, 6H), 1.86 (br s, 4H), 1.29 (t, 0 = 7.0 Hz, ZN), 1.28 (t, 0 = 7.0 Hz, ZN) ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.81 (br s, 1H), 7.98 (s, 1H), 7.97 (br s, 1H), 7.76 (dd , 0 = 8.3, 1.5 Hz, 1H), 7.52 (d, 0 = 8.3 Hz, 2H), 7.34 (br s, 1H), 7.24 (t, 0 = 7.9 Hz, 1H), 7.16 (d, 0 = 8.0 Hz, 1H), 6.59 (s, 2H), 4.43-4.59 (m, 4H), 4.2 4 (br s, 4H), 3.14 (s, 3H), 2.11 LC-MS Method N retention time = 6.58 4 min, [M + H] ⁺ = 7 07.2

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Example

129

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (1hydroxyethyl) -1Hbenzo [d] imidazole -1yl) butyl) -1Нbenzo [d] imidazole-5carboxamide

Way

(lat. s, ZN), 2.10 (s, 3H), 1.85 (br s, 4 H), 1.66 (s, ZN), 1.27 (m, 6 H) ^X H NMR (4 00 LC-MS MHz, DMSO-dg) method D: δ ppm 12.83 time (s, 1H), holding 12.23 (s, nia = 0.95 1H), 7.92- MIN, 8.02 (m, 2H), [M + H] ⁺ = 6 7.76 (dd, J = 8.3, 1.3 Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.34 (br s, 1H), 7.17 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 7.5 Hz, 1H), 6.61 (s, 1H), 6.58 (s, 1H), 5.88 (d, J = 3.3 Hz, 1H), 5.11 (d, J = 4.3 Hz, 1H), 4.56 (m, 4H), 4.27 (m, 4H), 2.10 (d, J = l, 3 Hz, 6H), 1.86 (br s, 4H), 1.42 (d, J = 6.5 Hz, ZN), 1.29 (q, J = 7.3 Hz, 6 N) 80.6

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Example

130

1- (4- (4- (2-amino-Pmethylacetamido) -2- (1ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo [d] imidazole-1yl) butyl) -2- (1- ethyl-Zmethyl-1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-5carboxamide trifluoroacetate

ABOUT

Way

Example

131

2- (1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido method) - 11

1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4 (methylsulfonamido) -1Hbenzo [d] imidazole-1yl) butyl) -1H-

^X H NMR (4 00 LC-MS MHz, DMSO-dg) method D: δ ppm 12.81 time (br. s, 1H), holding 8.16 (br. S, nia = 0.7 8 1 Η), 7.99 MIN, (s, 2 Η), [M + H] ⁺ = 7 7.92 (br s, 2H), 7.77 (dd, J = 8.4, 1.4 Hz, 1H), 7.64 (d, J = 7.8 Hz, 1H), 7.48- 7.59 (m, 1H), 7.35 (m, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.23- 7.29 (m, 1H), 6.68 (br s, 1H), 6.61 (s, 1H), 4.56 (q, J = 6.9 Hz, 4 Η), 4.46 (br. S, 2), 4.26 (br s, 4H), 3.24 (s, 3H), 2.08-2.15 (m, 6H), 1.86 (br s, 4H), 1.19-1.35 (m, 6H) 22.6 ^X H NMR (4 00 LC-MS MHz, flMCO-d ₆ ) method D: δ ppm 10.37 time (s, 1H), 7.98 holding (d, J = l, 3 Hz, nia = 0.90 1H), 7.96 MIN, (br. s, 1H), [M + H] ⁺ = 7 7.76 (dd, 29.6

- 170 036435

benzo [d] imidazole-5carboxamide trifluoroacetate О О X О ^Н Г Н у— ⁿ ' _N hn — ΊΓ j | Λ Η Г /> - ^ΝΗ I're .. WyMV uch, JL J = 8.5, 1.5 Hz, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.34 (br s, 1H), 7.31 (dd, J = 7.3, 1.5 Hz, 1H), 7.17-7.27 (m, 2 H), 6.61 (s, 1H), 6.59 (s, 1H), 4.51-4.61 (m, 4H), 4.16- 4.34 (m, 4H), 3.04 (s, 3 H), 2.10 (s, ZN), 2.09 (s, ZN), 1.86 (br s, 4H), 1.30 (td, J = 7.2, 2.8 Hz, 6H) Example 132 1- (4- (4- (2- (dimethylamino) -Nmethylacetamido) -2- (1ethyl-3-methyl-1H-pyrazole5-carboxamido) -1Hbenzo [d] imidazole-1yl) butyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) - 1Нbenzo [d] imidazole-5carboxamide tristrifluoroacetate Way eleven ^X H NMR (4 00 MHz, DMSO- ₆ ) δ ppm 12.8 (br s, 1H), 9.50 (br s, 1H), 7.99 (s, 2H), 7.78 ( dd, P = 8.4, 1.4 Hz, 1H), 7.62 (d, J = 1.8 Hz, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7, 35 (br s, 1H), 7.21-7.33 (m, 2H), 6.62 (s, 2H), 4.52-4.63 (m, 6H), LC-MS Method D Retention Time = 0.7 9 min, [M + H] ⁺ = 7 50.7

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and ¹ \ YES ^ N. o H y - ^N 'N hn - &] Π) Α ^ γ ¹⁴ Λ || 1 /> - ^ΝΗ ν, ... Η ₂ ΝX / Ν о о 4.28 (m, 4Η), 3.23 (s, 3N), 2.70 (d, 0 = 4.0 Hz, 6H), 2.11 (d, 0 = 1.5 Hz, 6H), 1.87 (br. s, 4H), 1.28 (db, 0 = 14.1, 7.0 Hz, 6H) Example 133 1- (4- (4- (2- (dimethylamino) acetamido) 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butyl) -2- (1 -ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide tristrifluoroacetate jj 1 DE ^ N. 11 y - ⁿ -n hn - 1 Π ν Λ H G ζ> - ^ΝΗ ν, ... ^Η2Ν ν ^Λ ^ ^Ν Way eleven ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.82 (brs, 1H), 12.7 (brs, 1H), 10.91 (s, 1H), 9.94 ( br s, 1H), 7.97 (s, 2H), 7.76 (d, 0 = 8.3 Hz, 1H), 7.72 (d, 0 = 8.0 Hz, 1H), 7, 52 (d, 0 = 8.5 Hz, 1H), 7.34 (d, 0 = 7.8 Hz, 2H), 7.18-7.28 (m, 1H), 6.60 (s, 1H ), 6.59 (s, 1H), 4.55 (q, 0 = 6.8 Hz, 4H), 4.27 (br s, 6H), 2.92 (s, 6H), 2.10 (s, 6H), 1.87 (br s, 4H), 1.29 (t, 0 = 7.0 Hz, 6H) LC-MS Method D Retention Time = 0.8 0 min, [M + 2H] ⁺ / 2 = 369.0 Example 2- (1-ethyl-3-methyl-1H- Way ^X H NMR (4 00 LC-MS

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134 pyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (2hydroxyacetamido) -1Hbenzo [d] imidazole-1yl) butyl) -1H-benzo [d ] imidazole-5carboxamide bistrifluoroacetate О Η р ^Ν 'Ν ην— / 7 Π) Il Г /> - ^ΝΗ Ν, ... ο ο eleven MHz, DMSO-dg) δ ppm 12.83 (br s, 1H), 12.52 (br. s, 1H), 10.32 (br s, 1H), 7.99 (s, 1H), 7.97 (br s, 1H), 7.76 (d, 0 = 8.3 Hz, 1H), 7.54 (d, 0 = 8.3 Hz, 2H), 7.34 (width s, 1 H), 7.30 (d, 0 = 8.0 Hz, 1H), 7.16- 7.23 (m, 1H), 6.61 (s, 2H), 4.50-4.62 (m, 4H), 4.27 (br s, 4H), 4.14 (s, 2H), 2.68 (br s, 1H), 2.11 (s, 6H), 1.87 (br. s, 4H), 1.30 (q, 0 = 7.0 Hz, 6H) method D: retention time = 0.8 6 min, [M + 2H] ⁺ / 2 = 355.4 Example 135 2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5- carboxamido) -4- (L / -methyl2- (methylamino) acetamido) 1H-benzo [d] imidazole-1- yl) butyl) -1H- benzo [d] imidazole-5carboxamide tristrifluoroacetate Way eleven ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.83 (br s, 1H), 8.61 (br s, 1H), 7.918.04 (m, 2 H), 7.77 ( d, 0 = 8.5 Hz, 1H), 7.63 (br s, 1H), 7.44-7.59 (m, 2H), 7.19- LC-MS Method D Retention Time = 0.7 9 min, [M + 2H] ⁺ / 2 = 369.0

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O II n O n t ⁿ -n hn- ('] Η Ν Ά Η Γ ζ> ~ ^NH N, ... ν-ς 8 θ 7.40 (m, ZN), 6.55-6.64 (m, 2H), 4.48- 4.63 (m, 4H), 4.26 (br s, 6H), 3.33 (s, 1H), 3.24 (s, 2H), 2.63- 2.71 (m, 1H), 2.44 (t, J = 5, l Hz, 2H), 2.05- 2.19 (m, 6H), 1.76-1.95 (m, 4H), 1.28 (q, J = 6.7 Hz, 6H) Example 136 1- (4- (4- (aminomethyl) -2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butyl) -2- (1-ethyl-Zmethyl- 1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-5carboxamide tristrifluoroacetate nh ₂ \ ^ \ o | H y - \ Ν'-n ⁷ hn-C ΊΓ H N \ Il Г Z> ~ ^NH Ν'-m \ / N 8 θ Way eleven ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.82 (br s, 1H), 12.53 (br s, 1H), 8.05 (br s, 3H), 7.97 (s, 2H), 7.76 (d, J = 8.3 Hz, 1H), 7.55 (t, J = 8.5 Hz, 2H), 7.25-7.39 (m, 3H) , 6.61 (s, 2H), 4.56 (d, J = 7.0 Hz, 4H), 4.38 (m, 2H), 4.27 (br s, 4H), 2.10 ( s, 6H), 1.84 (br s, 4H), 1.21-1.36 (m, 6 H) LC-MS Method D Retention Time = 0.7 6 min, [M + 2H] ⁺ / 2 = 333.4 Example 2- (1-ethyl-3-methyl-1H- Way ^X H NMR (4 00 LC-MS

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137

Example

138 pyrazole-5-carboxamido) - 11

1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (2 (methylamino) acetamido) 1H-benzo [d] imidazol-1yl) butyl) -1Hbenzo [d] imidazole- 5carboxamide tristrifluoroacetate

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (2hydroxy-ΛΖmethylacetamido) -1Нbenzo [d ] imidazole-1yl) butyl) -1Нbenzo [d] imidazole-5carboxamide bis

MHz, DMSO-dg) δ ppm 12.80 (br s, 1H), 12.54 (br. s, 1H), 10.86 (br s, 1H), 8.88 (br s, 2H), 7.97 (s, 2H), 7.76 (d, 0 = 8.0 Hz, 1H), 7.69 (d, 0 = 8.0 Hz, 1H), 7.53 (d, 0 = 8.3 Hz, 1H), 7.33 (d, 0 = 7.8 Hz, 2H), 7.19- 7.27 (m, 1H), 6.59 (s, 2H), 4.55 (q, 0 = 7.0 Hz, 4 N), 4.26 (m, 4H), 4.08 (t, 0 = 5.5 Hz, 2H), 2.63- 2.72 (m, ZN), 2.10 (s, 6 H), 1.87 (br. s, 4H), 1.29 (t, 0 = 7.0 Hz, 6H) method D: retention time = 0.7 9 min, [M + 2H] ⁺ / 2 = 362.0 ^X H NMR (4000 MHz, DMSO-dg) δ ppm 12.83 (br s, 2H), 7.99 (d, 0 = 1.3 Hz, 1H), 7.96 (br s, 1H ), 7.76 (dd, 0 = 8.3, 1.5 Hz, 1H), 7.54 LC-MS Method D Retention Time = 0.8 6 min, [M + 2H] ⁺ / 2 = 362.4

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trifluoroacetate O \ ^ ^on x ^ o H y - / ^N -N HN - &] H Ά II D)> - ^{NH n} -m y / Ν 8 o (br s, 2H), 7.33 (br s, 1H), 7.26 (t, 0 = 7.9 Hz, 1H), 7.19 (d, 0 = 7.3 Hz, 1H), 6.61 (br s, 2H), 4.45-4.63 (m, 4H), 4.25 (br s, 4H), 3.44-3.86 (br s, 2H), 3.29 (br s, 1H), 3.18 (br. s, ZN), 2.11 (br. s, ZN), 2.10 (br s, 3H), 1.86 (br s, 4H), 1.21- 1.35 (m, 6H) Example 139 1- (4- (4- (2aminoacetamido) -2- (Ιο tyl-3-methyl-1H-pyrazole 5-carboxamido) -1Нbenzo [d] imidazole-1- yl) butyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide Way eleven ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.82 (br s, 1H), 12.6 (br s, 1H), 10.81 (s, 1H), 8.20 (br s, 3H), 7.97 (d, 0 = 1.3 Hz, 2H), 7.76 (dd, 0 = 8.5, 1.5 Hz, 1H), 7.69 (d, 0 = 8.0 Hz, 1 H), 7.53 (d, 0 = 8.5 Hz, 1H), 7.33 (d, 0 = 8.0 Hz, 2H), 7.19- LC-MS Method D Retention Time = 0.7 9 min, [M + H] ⁺ = 7 08.6

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О JL ^ NH, О Η 2 ⁿ 'n hn - & IH) A Il T /) - ^NH N, ... w J? oh oh 7.28 (m, IH), 6.59 (s, 2 Η), 4.55 (q, 3 = 7.0 Hz, 4Η), 4.26 (d, 3 = 6.3 Hz, 4Η), 3.85- 4.02 (μ, 2Η), 2.10 (s, 6Η), 1.87 (br. S, 4Η), 1.29 (τ, 3 = 7.2 Hz, 6Η) Example 140 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) - 1- (4- (2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -4- (5oxoimidazolidine-1 -yl) 1Н-benzo [d] imidazol-1- yl) butyl) -1Н-benzo [d] imidazole-5carboxamide HN— \ Η у— ^N N HN— ('] Π>Ar' ^X5 X-- ^N A Η TZ> - ^{NH N} -M ivkAAiT ν-ς ([o o x-H Way eleven ^Χ Η NMR (4 00 MHz, DMSO-dg) δ ppm 12.7 (br s, 1Η), 12.4 (br s, 1H), 7.96 (s, 2H), 7.74 (dd , 3 = 8.4, 1.3 Hz, 1H), 7.51 (d, 3 = 8.6 Hz, 1H), 7.38 (d, 3 = 8.1Hz, 1 Η), 7.35 (br s, 1H), 7.23 (t, 3 = 8.1 Hz, 1 H), 6.98 (d, 3 = 7.8 Hz, 1H), 6.61 (s, 1H), 6.59 (s, 1H), 4.86 (br s, 2H), 4.4 6-4.64 (m, 4H), 4.26 (br s, 4H), 3.73 (br s, 1H), 3.55 (br s, 2H), 2.10 (s, 6H), LC-MS Method E Retention Time = 0.7 3 min, [M + H] ⁺ = 7 20.9

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Example

141

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (3-methyl5-oxoimidazolidine-1- yl) 1Н-benzo [d] imidazole-1yl) butyl) -1Нbenzo [d] imidazole-5carboxamide

Way

Example

142

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (3methylbutanamido) -1HC

1.87 (br. S, 4H), 1.21- 1.38 (m, 6H) ^T H NMR (400 MHz, DMSO- ₆ ) δ ppm 12.81 (br s, 1H), 12.24 (br s, 1H), 7.97 (s, 2 H), 7.75 (dd, J = 8.4, 1.3 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H), 7.41 (d, J = 8, l Hz, 1H), 7 , 34 (br s, 1H), 7.25 (t, J = 8, l Hz, 1 H), 7.00 (d, J = 7.8 Hz, 1H), 6.61 (s, 1H ), 6.59 (br s, 1H), 4.66 (br s, 2H), 4.48-4.61 (m, 4H), 4.27 (br s, 4H), 3, 51 (br s, 2H), 2.46 (s, 3H), 2.10 (s, 6H), 1.80-1.93 (m, 4H), 1.22-1.36 (m, 6H) LC-MS Method D Retention Time = 0.82 min, [M + H] ⁺ = 7 34.6 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.82 (br s, 1H), 12.53 (s, 1H), 10.36 lx-ms method E: retention time = 0 _g 99 min,

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benzo [d] imidazole-1- (s, 1H), [M + H] ⁺ = 7 yl) butyl) -1Нbenzo [d] imidazole-5carboxamide O | o Pu- ⁿ -n hn - ((] Π ₍ ^χΖ5 ^ Γ- ^Ν \ Il T ^NH N, ... J o '-X 7.91-8.01 (m, 2H), 7.76 (dd, C = 8.4, 1.3 Hz, 1H), 7.65 (d, J = 8, l Hz, 1H), 7.54 (d, J = 8.6 Hz, 1H), 7.34 (br s, 1H), 7.22-7.30 (m, 1H), 7.12- 7.21 (m, 1H), 6.59 (s, 1H), 6.58 (s, 1H), 4.56 (q, J = 1.0 Hz, 4H), 4.18- 4.32 (m, 4H), 2.33 (d, J = 7, l Hz, 2H), 2.15 (m, 1H), 2.09 (s, 6H), 1.86 (br. s, 4H), 1.29 (td, J = 7.0, 4.6 Hz, 6H), 0.98 (d, J = 6.6 Hz, 6H) 36, 1 8-ethyl-Yu, 18-dimethyl- Way ^X H NMR (4 00 LC-MS 7,20-DIOXO-23- (5- 13 MHz, DMSO-dg) method C: oxoimidazolidin-l-yl) - δ ppm 8.02 time 6,7,8,11,12,13,14,15,20,2 (s, 1H), 7.98 holding 1,28,29,30,31- (br. s, 1H), nia = 0.83 tetrahydrobenzo [4,5] - 7.82 (d, MIN, imidazo [1,2- J = 8.5 Hz, [M + H] ⁺ = 7 a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'- 1] [1,3,6,15,17] pentaazacy clogeneicosin-3- 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.52 (d, J = 8.0 Hz, 60.4

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carboxamide n O _{H; N} AfY ^N 'ML U I / ^-NH / ⁴ X I! JL /> ~ \ ⁿ / ⁿ 'n ° x> ° * —NH 1H), 7.287.40 (m, 2 Η), 7.06 (br s, 1H), 6.55 (br s, 1H), 4.87 (br s, 2H), 4.74 (br s, 2H), 4.47 (q, J = 7.3 Hz, 2 Η), 4.2 4 (br s, 4H), 3.55 (br s, 2H), 2.82 (br s, 2H), 2.16 (s, 3H), 2.09 (s, 3H), 1.92 (br s, 4H), 1.81 (br s, 2H), 1.49 (br s, 2H), 1.38 (br s, 2H), 1.30 (t, J = 6.9 Hz, 3H) Example 144 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) - 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (2 (methylamino) ethoxy) -1Hbenzo [d] imidazole-1- yl) butyl) -1H- benzo [d] imidazole-6carboxamide Way eleven ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 12.81 (br s, 1H), 11.98 (br s, 1H), 8.90 (br s, 2H), 7.97 (s, 2H), 7.76 (dd, J = 8.3, 1.5 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H), 7.35 (br s, 1H), 7.14-7.22 (m, 2H), 6.91 (dd, D = 6.1, LC-MS Method C Retention Time = 0.7 9 min, [M + H] ⁺ = 7 09.4

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Example 2- (1-ethyl-3-methyl-1H- 146 pyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -4-methyl-1Hbenzo [d] imidazol-1-yl) butyl) -1H-benzo [d] imidazole-5carboxamide ditrifluoroacetate ° 1 H y - ^ν -ν ην - & I Π) A ³ H I /> - ^NH N, ... X / N Π о VA

Example

147 methyl 2 - ((1- (4- (5carbamoyl-2- (1-ethyl-Zmethyl-1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-1-

2H) 2.82 (d, J = 4.65 Hz, 4 Η) 2.74-2.79 (m, IH) 2.16 (s, 3H) 2.08 (s, 3H) 1.89 (br s, 4H) 1.80 (br s, 2H) 1.48 (br s, 2H) 1.37 (br s, 2H) 1.30 (t, J = 6.97 Hz, ZN) Way eleven ⁷ H NMR (4000 MHz, DMSO-dg) δ ppm 1.29 (m, 6H) 1.85 (br s, 4H) 2.07-2.19 (m, 6H) 4.25 (d, J = 7.83 Hz, 4H) 4.55 (quint, J = 6.69 Hz, 4H) 6.59 (s, 2H) 7.01-7.19 (m, 2H) 7.307.41 (m, 2H) 7.53 (d, J = 8.34 Hz, 1 H) 7.75 (dd, J = 8.46, 1.39 Hz, 1H) 7.928.05 (m, 2H) 12.83 (br . s, 2H) LC-MS Method C Retention Time = 0.98 min, [M + H] ⁺ = 6 50.5 Way eleven ^X H NMR (400 MHz, DMSO-dg) δ ppm 1.221.34 (m, 6H) 1.85 (br s, LC-MS method D: time retention = 1.03

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yl) butyl) -2- (1-ethyl-Zmethyl-1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-4yl) oxy) acetate ditrifluoroacetate О Н у— ^N -N hn— &] Η Il Γ JAnH N, ... y / J O 4H), 2.11 (d, 0 = 1.00 Hz, 6H), 3.94 (s, 3H) 4.24 (d, 0 = 9.29 Hz, 4H), 4.45- 4.60 (m, 4H), 6, 5 6-6, 64 (m, 2H), 6.90 (d, 0 = 7.78 Hz, 1H) 7.12-7.23 (m, 2H), 7.33 (br s, 1H), 7.51 (d, 0 = 8.53 Hz, 1H) 7.75 (dd, 0 = 8.53, 1.51 Hz, 1H), 7.97 (D, 0 = 1.51 Hz, 2H), 12.80 (br. s, 2H) min, [M + H] ⁺ = 6 66.5 Example 148 2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5- carboxamido) -4-methoxy1Н-benzo [d] imidazol-1yl) butyl) -1Н- benzo [d] imidazole-5carboxamide ditrifluoroacetate Way 16 ^X H NMR (400 MHz, DMSO-dg) δ ppm 1.29 (m, 6 Η) 1.86 (br. S, 4H) 2.11 (s, 6H) 3.71 (s, 3H) 4 , 25 (br s, 4H) 4.56 (q, 0 = 7.01 Hz, 4H) 6.58-6.65 (m, 2H) 5.00 (s, 2H) 6.88 (br. s, 1H) 7.12-7.22 (m, 2H) 7.30 (br s, 1H) 7.51 (d, 0 = 8.31 Hz, 1H) 7.75 (dd, LC-MS Method D retention time = 0.97 min, [M + H] ⁺ = 7 24.5

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Example

149

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (2hydroxyethoxy) -1

N-benzo [d] imidazole-1yl) butyl) -1Hbenzo [d] imidazole-5carboxamide diitrifluoroacetate

Way

0 = 8.44, 1.59 Hz, 1H) 7.908.00 (m, 2H) 12.79 (br. s, 2H) ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 1.2 ΟΙ, 39 (m, 6Η) 1.85 (br s, 4Η) 2.10 (d, 0 = 1.96 Hz, 6H) 3.74-3.82 (m, 3H), 4.13 (t, 0 = 4.52 Hz, 2H), 4.20-4.31 (m, 4H), 4.48-4.61 ( m, 4H), 6.60 (s, 2H), 6.87 (d, 0 = 7.58 Hz, 1H), 7.10-7.20 (m, 2H), 7.34 (br. s , 1H), 7.53 (d, 0 = 8.31 Hz, 1H), 7.75 (dd, 0 = 8.56, 1.47 Hz, 1H), 7.97 (d, 0 = 1, 47 Hz, 2H), 12.11 (br s, 1H) 12.83 (br s, 2H) LC-MS Method D Retention Time = 0.90 min, [M + H] ⁺ = 6 96.4

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Example 150 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4- (hydroxymethyl) -1Hbenzo [d] imidazole -1yl) butyl) -1Нbenzo [d] imidazole-5carboxamide ditrifluoroacetate ОН у - ^ν 'ν ην-1 II) A Η 1 JA ^nh о о Way eleven ^X H NMR (4000 MHz, DMSO-dg) δ ppm 1.241.35 (m, 6H), 1.87 (br s, 4H), 2.00-2.14 (m, 6H), 4.27 (d, J = 4.65 Hz, 4H), 4.56 (quint, J = 7.21 Hz, 4H), 4.83 (s, 2H) 6.60 (d, D = 8.56 Hz, 2H), 7, Οδ- Ο, 12 (m, 1H) 7.14-7.23 (m, 1H), 7.31 (br s, 1H), 7.41 (d, J = 7.83 Hz, 1H), 7.53 (d, J = 8.31 Hz, 1H), 7.75 (dd, J = 8.56, 1.47 Hz, 1H), 7.89-8.01 (m , 2 H), 12.17 (br s, 1H) 12.80 (br s, 2H) LC-MS Method D Retention Time = 0.8 9 min, [M + H] ⁺ = 6 66.5 Example 151 1- (4- (4- (2- (dimethylamino) -2- oxoethoxy) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-1yl) butyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5- Way 16 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 1.181.38 (m, 6H), 1.80-1.94 (m, 4Η), 2.11 (s, 6Η), 2.85 (s , 3 Η), 3.00 (s, 3 Η), 4.26 (d, LC-MS Method E Retention Time = 0.8 8 min, [M + H] ⁺ = 7 37.5

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carboxamide 1 O P 7— \ ⁿ 'n hn — P J Π N G /> - ^NH N, ... \ / N O ° J = 10.27 Hz, 4H), 4.49- 4.64 (m, 4H), 5.02 (s, 2H), 6.60 (d, J = 9.29 Hz, 2H) 6.81-6.92 (m, 1H), 7.10-7.20 (m, 2 H) 7.30 (br s, 1H), 7.52 (d, J = 8.31 Hz, 1H), 7.75 (dd, J = 8.44, 1.35 Hz, 1H), 7.87-8.03 (m, 2H) 12.09 (br s, 1H), 12.80 (s, 1 H) Example 152 1- (4- (4- (2-amino-2oxoethoxy) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-1- yl) butyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1H-benzo [d] imidazole-5carboxamide ditrifluoroacetate Way 16 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 1.261.33 (m, 6H), 1.86 (br s, 4H), 2.07-2.14 (m, 6H), 4.25 (d, J = 9.54 Hz, 6H), 4.514.60 (m, 4H) 6.59 (d, J = 7.09 Hz, 1H), 6.86 (dd, J = 5.62, 3 , 42 Hz, 1H), 7.13-7.16 (m, 2H), 7.30 (br s, 1H), 7.50-7.59 (m, 2H), 7.76 LC-MS Method D Retention Time = 0.8 6 min, [M + H] ⁺ = 7 09.4

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° <sx ^NH2 О ° Н у - ^ν 'ν ην - 1 Η? (γ ASsy.-N Ύ Il Τ Ά ^{ΝΗ ν -ν} μ j 8 ° (dd, 0 = 8.31, 1.47 Hz, 1Η), 7.89-8.01 (m, 2H), 8.44 (br s, 1H), 12.77 (br. s, 2H) Example 153 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -4- (2 (methylamino) -2oxoethoxy ) -1Нbenzo [d] imidazol-1yl) butyl) -1Нbenzo [d] imidazole-5carboxamide 1 ° \ / ^NH о γ П у-4 ^Ν Ν ΗΝ-ά 'l · Η ίί Г /> ~ ^{ΝΗ ν} - ~ μ J? 8 ο Way 16 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 1.251.35 (m, 6), 1.86 (br. S, 4Η), 2.04-2.18 (m, 6Η), 2, 75 (d, 0 = 4.40 Hz, ZN), 4.25 (br. S, 4Η), 4.52-4.63 (μ, 7Η), 6.54-6.63 (m, 2Η) , 6.84-6.90 (μ, 1Η), 7.16 (d, 0 = 4.65 Hz, 2Η), 7.31 (br.s, 1Η), 7.53 (d, 0 = 8 , 31 Hz, 1H), 7.76 (d, 0 = 8.31 Hz, 1H), 7.90-8.01 (m, 2H), 8.64 (d, 0 = 4.40 Hz, 1H ), 12.31 (br s, 1H), 12.80 (br. LC-MS Method D Retention Time = 0.8 9 min, [M + H] ⁺ = 7 23.5

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s, 1H) Example 154 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1Н-pyrazole-5-carboxamido) -1Нbenzo [d] imidazol-1-yl) butyl) -2- (1-ethyl-Zmethyl- 1Н-pyrazole-5carboxamido) -N- (2morpholinoethyl) -1Нbenzo [d] imidazole-5carboxamide tritrifluoroacetate 0 h ₂ n – V ( ^Η 1Ω ^ν S θχχ '/' Ν fi [ΝΗ ΝΗ Way eleven ^X H NMR (400 MHz, DMSO-dg) δ ppm 9.66 (br s, 1H), 8.70-8.77 (m, 1H), 7.98 (s, 3H) 7.68- 7.81 (m, 2H), 7.57 (dd, J = 19.20, 8.34 Hz, 2H), 7.38 (br s, 1H), 6.61 (d, J = 9, 60 Hz, 2H), 4.51- 4.66 (m, 4H), 4.29 (br s, 4H), 4.03 (d, J = ll, 87 Hz, 2H), 3.54- 3.75 (m, 6H), 3.35 (br s, 2H), 3.16 (d, J = 9.60 Hz, 2H), 2.07-2.13 (m, 1H), 2 , 11 (s, 6H), 1.88 (br s, 4H), 1.31 (m, J = 7.07 Hz, 6H) LC-MS Method C Retention Time = 0.7 5 min, [M + H] ⁺ = 7 92.7 Example 155 1- (4- (5-carbamoyl-2- (1ethyl-3-methyl-1Η-pyrazole-5-carboxamido) -1Нbenzo [d] imidazole-1- yl) butyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -Ν, Νdimethyl-1H- benzo [d] imidazole-5carboxamide Way eleven ^X H NMR (4 00 MHz, DMSO- ₆ ) δ ppm 12.7312.87 (m, 2Η), 7.97 (s, 1Η), 7.75 (d, J = 8.34 Hz, 1H) , 7.48-7.58 (m, 3H), 7.34 (br s, 1H) 7.24 (dd, LC-MS Method C Retention Time = 0.8 6 min, [M + H] ⁺ = 7 07.7

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O H ₂ N-Y v xjl ° ^<n x> 7 ^Ο \\ ^ Ν / y J1 J = 8.34, 1.26 Hz, 1H), 6.60 (d, J = 9.60 Hz, 2H) 4.56 (D, J = 7.07 Hz, 4H), 4.27 (D, J = 6.32 Hz, 4H), 2.95 (br. S, J = 15.66 Hz, 6H), 2.09 (d, J = l, 77 Hz, 6H), 1.87 (br. s, 4H), 1.30 (m, σ = Ί, 07 Hz, 6H), 1.23 (s, 1H) Example 156 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole 5-carboxamido) -1Nbenzo [d] imidazol-1- yl) butyl) -2- (1-ethyl-Zmethyl-1H- pyrazole-5carboxamido) -N-methyl-1Hbenzo [d] imidazole-5carboxamide diitrifluoroacetate O H ₂ N - & ^ \ JL ° < ⁿ S ° W ^ ^N 'N H G - ^NH θ ^ γΑ ^ Α-Ν ⁷ ^ ΝΗ Way eleven ^X H NMR (400 MHz, DMSO- ₆ ) δ ppm 12.84 (br s, 2H), 8.41 (d, J = 4.55 Hz, 1H), 7.95 (d, J = 14.15 Hz, 3H), 7.75 (d, J = 8.34 Hz, 1H), 7.69 (d, J = 8.34 Hz, 1H), 7.54 (dd, J = 8, 46, 2.65 Hz, 2H), 7.34 (br s, 1H), 6.59 (s, 2H) 4.56 (q, J = 6.99 Hz, 4H), 4.27 (br s, 4H), 2.79 (d, s7 = 4.2 9 Hz, 3H), 2.09 (s, LC-MS Method C Retention Time = 0.8 0 min, [M + H] ⁺ = 6 93.6

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Example

157

2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -7isopropoxy-1Hbenzo [d] imidazole-1yl) butyl ) -1Нbenzo [d] imidazole-5carboxamide h ₂ n

Way

Example 23- (2- (dimethylamino) -N- Way 158 methylacetamido) -8-ethyl 10,18-dimethyl-7,20- dioxo 6,7,8,11,12,13,14,15,20,2 1,28,29,30,31- tetrahydrobenzo [4,5] imide zo [1,2- a] benzo [4,5] imidazo [2,1- 13

6H), 1.86 (br. s, 4H), 1.29 (t, 0 = 6.95 Hz, 6H) ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.78 (br s, 2H), 8.02-7.92 (m, 2H), 7.75 (dd, 0 = 8.4, 1.3 Hz, 1H), 7.53 (d, 0 = 8.4 Hz, 1H), 7.32 (br s, 1H), 7.14-7.03 (m, 2H), 6, 85- 6.75 (m, 1H), 6.59 (s, 2H), 4.74-4.63 (m, 1H), 4.62- 4.51 (m, 4H), 4.43- 4.22 (m, 4H), 2.11 (s, 3H), 2.10 (s, 3H), 1.97-1.69 (m, 4H), 1.35-1.23 (m, 6H), 1.15 (d, 0 = 6.0 Hz, 6H) LC-MS Method A retention time = 1.51 8 min, [M + H] ⁺ = 6 94.5 ^X H NMR (4 00 MHz, (methanol-dj δ 8.35 (s, 1H), 7.99 (s, 2H), 7.88 (s, 1H), 7.72 (d, 1H), 7 , 46 (br s, 2H), 7.35 (d, LC-MS Method A retention time = 0.82 min, [M + H] ⁺ = 7 90.6

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p] dipyrazolo [5,1-e: 4 ', 3' 1] [1,3,6,15,17] - pentaazacyclogenaicosin-3-carboxamide trifluoroacetate OH ₂ 1 \ 1 - Pt 5-Λ (G 'G 'Y ^N ₄ N Χ ^' - Ν Λ- (Г Ν I о ^{/ А} м n X <\। о 1H), 6.75 (s, 1H), 4.47- 4.62 (m, 3H), 4.35 (br s, 6H), 3.71 (br s, 2H), 3.43 (s, 2H), 3.12 (s, 1H), 2.79-2.92 (m, 6H), 2.29 (d, J = 6.3 Hz, 2H), 2.20 (br s, 4H), 2.05 (br s, 4H), 1.90 (br s, 6H), 1.62 (br s, 2H), 1.32- 1.45 (m, 6H) Example 159 23- (2-amino-Pmethylacetamido) -8-ethyl 10,18-dimethyl-7,20 dioxo 6,7,8,11,12,13,14,15,20,2 1,28,29,30,31- tetrahydrobenzo [4,5] imide zo [1,2- a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3' 1] [1,3,6,15,17] - pentaazacyclogeneicosine 3-carboxamide Way 13 ^X H NMR (4 00 MHz, DMSO-dg) δ ppm 7.958.07 (m, 2H), 7.88-7.15 (m, 5Η), 6.75 (bs, IH), 4.69 (br s, 2H), 4.35 (br s, 6H), 4.49 (d, J = 7, l Hz, 2H), 4.22 (br s, 6H), 3.34 ( s, 3H), 2.75-2.89 (m, 2H), 2.21 (s, 2H), 2.17 (s, 2H), 1.91 (br s, 3H), 1.72 -1.85 (m, 3H), 1.50 (br s, 2H), LC-MS Method A retention time = 0.81 min, [M + H] ⁺ = 7 62.7

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yl) but-2-en-1-yl) -7- (suapomethyl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide O h ₂ n ~ ^ fl ' λ '// ^ο Ν— \ U / Η Γ} _ \ ^ - ^Ν ' Ν NC // __ / / O /, Χ— N h ₂ n Л x у \ h VXX NN II NH. ABOUT ---' (lat. s, 2Η), 8.12-7.80 (m, 4Η), 7.73 (d, J = 6.5 Hz, 2Η), 7.48- 7.30 (m, ZN), 6.55 (s, 2Η), 6.01-5.94 (m, IH), 5.62- 5.54 (μ, 1Η), 5.01 (s, 2Η), 4.82 (s, 2Η), 4.56-4.46 (μ, 4Η), 4.34 (s, 2Η), 2.11 (s, ZN), 2.08 (s, ZN), 1.29- 1.23 (m, 6Η) niya = 1.2 9 min, [M + H] ⁺ = 7 16, 4 Example 162 (E) -1- (4- (5-carbamoyl-2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) IH-benzo [d] imidazol-1yl) but-2-en-1-yl) -2- (1ethyl-3-methyl-1Н-pyrazole5-carboxamido) -7 (hydroxymethyl) -1Нbenzo [d] imidazole-5carboxamide О fl 4ji // (Η \ ν V] / Ν HO C 1 1 0 Η ₂ Ν J!, Χ / λ H Ν-Ν II Ν π ι Ο --- ' Method 9 ^τ Η NMR (4 00 MHz, flMCO-d ₆ ) δ ppm 12.92 (br s, 2Η), 8.02-7.91 (m, 4Η), 7.72 (d, J = 10, l Hz, 2H), 7.71 (s, IH) 7.43 (d, J = 8.4 Hz, IH), 7.34 (br s, 2H), 6.52 (d, J = 6, 7 Hz, 2H), 5.98 (d, J = 15.5 Hz, 1H), 5.51 (dd, J = 13.2, 8.0 Hz, 1H), 5.09 (s, 2H) , 4.81 (d, J = 4.2 Hz, 2H), 4.61 (s, 2H), 4.55- LC-MS Method A retention time = 1.23 4 min, [M + H] ⁺ = 7 07.9

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4.48 (m, 4H), 2.12 (s, 3H), 2.11 (s, 3H), 1.29-1.25 (m, 6H) Example 163 (E) -2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4 (morpholinomethyl) -1Hbenzo [ d] imidazol-1-yl) but-2-en-1-yl) -1Нbenzo [d] imidazole-5carboxamide O ^ ^NH 2 9 O. Г>'^ NH / \ N-y L /)>n' \ | / 1 = N / HN N / ^ O \ / Ν Ί Way eleven ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.90 (br s, 2H), 9.81 (s, 1H), 7.97 (s, 2H), 7.72-7.23 (m, 6H), 6.56 (d, 3 = 15.7 Hz, 2H), 5.93-5.85 (m, 2H), 4.85 (br s, 4H), 4.72 ( br s, 2H), 4.54-4.50 (m, 4H), 3.8 (br s, 2H), 3.5 (br s, 2H), 3.26 (br s, 4H), 2.13 (s, 6H), 1.30-1.22 (m, 6H) LC-MS Method A Retention Time = 1.30 8 min, [M + H] ⁺ = 7 33 Example 164 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) - 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -7 (morpholinomethyl) -1Hbenzo [d] imidazole-1- yl) butyl) -1H- benzo [d] imidazole-5carboxamide trifluoroacetate Way eleven ^T H NMR (400 MHz, DMSO-dg) δ ppm 12.84 (s, 2), 10.03 (s, 1), 7.97 (t, 3 = 4.4 Hz, 2), 7.75 (dd, 3 = 8.4, 1.3 Hz, 1), 7.66 (d, 3 = 7.6 Hz, 1), 7.51 (d, 3 = 8.4 Hz, 1), 7 , 32 (d, 3 = 12.3 Hz, LC-MS Method A Retention Time = 1.32 9 min, [M + H] ⁺ = 7 35.3

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Tl y - ν ^ν 'ν ην - {Ύ Η Α < ^ζΖ ^ γ- ^ ^Ν \ ίί Γ /> - ^{ΝΗ Ν} -κι S ° 3), 6.60 (τ, 0 = 11.2 Hz, 2), 4.52 (dd, 0 = 18.6, 13.6 Hz, 8Η), 4.25 (d, 0 = 6.2 Hz, 2H), 3.89 (s, 2), 3.61 (s, 2H, 3.25 (s, 4H, 2.08 ( d, 0 = 15.6 Hz, 6H), 1.81 (d, 0 = 22.1 Hz, 4H), 1.30 (t, 0 = 7.0 Hz, 6H) Example 165 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -7-methyl-1Hbenzo [d] imidazole -1yl) butyl) -1Н-benzo [d] imidazole-5carboxamide ° N y-H ^Ν -Ν ην— {1 Π) GT ιίΧΧ /) - NH Ν, ... \ / Ν 8 ° Way eleven ² H NMR (400 MHz, DMSO-dg) δ ppm 12.77 (s, 2H), 7.97 (t, 0 = 4.1 Hz, 2H), 7.76 (dd, 0 = 8.4, 1.4 Hz, 1H), 7.58 (d, 0 = 8.4 Hz, 1H), 7.37 (m, 2H), 7.09 (t, 0 = 7.7 Hz, 1H), 7 , 00 (d, 0 = 7.5 Hz, 1H), 6.58 (d, 0 = 21.8 Hz, 2H), 4.56 (dq, 0 = 14.2, 7.0 Hz, 4H) , 4.39 (t, 0 = 6.8 Hz, 2H), 4.28 (d, 0 = 6.9 Hz, 2H), 2.61 (d, 0 = 8.6 Hz, 3H), 2 , 09 (d, LC-MS Method A Retention Time = 1.47 0 min, [M + H] ⁺ = 6 50.3

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J = 14.8 Hz, 6H), 1.88 (dd, J = 29.7, 5.6 Hz, 4H), 1.29 (dt, J = 9.8, 7.1 Hz, 6H) Example 166 (E) -2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -4-methoxy1H-benzo [d ] imidazol-1yl) but-2-en-1-yl) 1Hbenzo [d] imidazole-5carboxamide \ __ _{_} O 11 y — x ⁿ 'n hn— {J Π \ nA / ¹ | ί 1 Α ^νη ν _% . , X / Ν 8 ° Way eleven ^X H NMR (400 MHz, DMSO- ₆ ) δ ppm 7.97 (m, 2 Η), 7.72 (d, J = 8.3 Hz, 1 H), 7.43 (d, J = 8 , 3 Hz, 1), 7.35 (s, 1 Η), 7.14 (t, J = 8, l Hz, 1 Η), 7.06 (d, J = 8.0 Hz, 1 Η ), 6.89 (s, 1 Η), 6.58 (m, 2 Η), 5.90 (s, 2 Η), 4.82 (m, 4H), 4.52 (m, 4 Η) , 3.94 (s, 3 Η), 2.14 (s, 6 Η), 1.301.24 (m, 6 H) LC-MS Method A Retention Time = 1.4 9 6 min, [M + H] ⁺ = 6 64.2 Example 167 (Ε) -2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1- (4- (2- (1-ethyl-3-methyl1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-1yl ) but-2-en-1-yl) 1Hbenzo [d] imidazole-5carboxamide Way eleven ⁷ H NMR (400 MHz, DMSO- ₆ ) δ ppm 12.76 (br.m, 2), 7.96 (m, 2), 7.71 (d, s7 = 8.5 Hz, 1 Η), 7.50 (s, 1 Η), 7.43 (d, J = 8.4 Hz, LC-MS Method A Retention Time NLI = 1.447 min, [M + H] ⁺ = 6 34.2

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o H y - ⁿ -n hn - jT y \ g ι ^χΖΪ Αγ ^ ^Ν \ ίί Γ JA ^NH Ν, ... ^η ₂ Ν ^! ^ Α _ν ^ y / Ν 8 ° 1H), 7.38 (s, 1 Η), 7.33 (s, 1 Η), 7.16 (m, 2H), 6.55 (d, 0 = 7.4 Hz, 2 Η), 5 , 91 (s, 2), 4.83 (s, 4), 4.53 (q, O = 7.1 Hz, 4), 2.13 (s, 6), 1.28 ( m, 6 N) Example 168 (Ε) -1- (4- (5-carbamoyl-2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) ΙΗ-benzo [d] imidazol-1yl) but-2-en-1-yl) -2- (1ethyl-3-methyl-1H-pyrazole5-carboxamido) -7 (morpholinomethyl) -1Hbenzo [d] imidazole-5carboxamide X ^ \ O ° N '// ^{n H} 2 -n hn — ς' 1 | l V-0 Η Г ^NH N_ .. J ο \ -Ύ Way 16 ^X H NMR (400 MHz, DMSO-dg) δ ppm 13.10 (br s, 2H), 8.01 (m, 4), 7.75 (m, 2 Η), 7.43 (m, 2 Η), 7.36 (s, 1 Η), 6.58 (s, 2 Η), 5.92 (d, 0 = 16.3 Hz, 1 H), 5.46 (d, 0 = 15 , 6 Hz, 1 Η), 5.11 (s, 2), 4.80 (s, 2), 4.54 (br. S, 6), 2.14 (s, 6 Η), 1.31-1.26 (m, 6 N) LC-MS Method A Retention Time NLI = 1.255 min, [M + H] ⁺ = 7 76.8 Example 169 The structure is not defined. One of two possible structures: 8-ethyl-10, 18-dimethyl7,2 0-dioxo Way 13 ^X H NMR (400 MHz, MeOG-d ₆ ) δ ppm 8.07 (d, J = l, 3 Hz, 1 Η), 7.89 lx-ms Method A: retention time NIJ

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6,7,8,11,12,13,14,15,20,2 1,28,29,30,31 tetrahydrobenzo [4,5] imidazo [1,2-a] benzo [4,5] imidazo [2 , 1-p] dipyrazolo [5,1-e: 4 ', 3'-L] [1,3,6,9,15,17] hexaazacyclogenicosine24-carboxamide N A ___. M 0 HN \ NH _N W nH / 'nA> \ = / A J Y 8-ethyl-10, 18-dimethyl-7,2 0-dioxo 6,7,8,11,12,13,14,15,20,2 1,28,29 , 30,31-tetrahydrobenzo [4,5] imide zo [1,2-a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'1] [1, 3,6,9,15,17] - hexaazacyclogenicosin-3carboxamide O HN \ NH nW jQj __J 0 HN y --- 'o ^ NH ₂ (dd, J = 8.6, 1.4 Hz, 1 H), 7.60 (d, J = 8.4 Hz, 1 H), 7.52 (d, J = 7.4 Hz, 1 H ), 7.31-7.44 (m, 3 H), 6.06 (s, 1 H), 4.87 (br s, 2 H), 4.26 (br s, 2 H), 4.20 (br s, 2 H), 4.08-4.16 (m, 2 H), 3.59 (t, J = 4.9 Hz, 2 H), 3.36-3.42 (m, 2 H), 3.08-3.14 (m, 2 H), 2.29 (s, 3 H), 2.21 (s, 3 H), 2.04 (br s, 4 H), 1.09 (t, J = 7, l Hz, 3 H) = 1.270 min, [M + H] ⁺ = 6 77.3 Example 170 The structure is not defined. One of two possible structures: Way 13 ^X H NMR (MeODd ₄ ) δ ppm 7.89-7.96 (m, LC-MS Method A: time

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8-ethyl-10, 18-dimethyl-7,2 0-dioxo 6,7,8,11,12,13,14,15,20,2 1,28,29,30,31-tetrahydrobenzo [4, 5] imide 3o [1,2-a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'- 1] [1,3,6,9,15, 17] - hexaazacyclogenicosine-24-carboxamide 0 HN I NH 'nA> \ = / А A ⁰ HN _z --J А N 8-ethyl-Yu, 18-dimethyl-7.2 0-dioxo-6,7,8 , 11,12,13,14,15,20,2 1,28,29,30,31-tetrahydrobenzo [4,5] imide zo [1,2-a] benzo [4,5] imidazo [2,1p ] dipyrazolo [5,1-e: 4 ', 3'- 1] [1,3,6, 9,15,17] - hexaazacyclogenicosine-3carboxamide _o ΗΝ | ΝΗ «/ b о ΗΝ> --- An Ο ^ ΝΗ ₂ 2 Η), 7.527.61 (m, ZN), 7.34-7.43 (μ, 2Η), 6.07 (s, 1Η), 4.854.94 (width μ, 2Η), 4.194.26 ( μ, 1Η), 4.13 (d, J = 7.3 Hz, 1Η), 3.57 (τ, J = 4.9 Hz, 2Η), 3.39 (τ, J = 6.3 Hz, 2H), 2.29 (s, ZN), 2.21 (s, ZN), 2.05 (br. S, 4H), 1.08 (t, J = 7, l Hz, ZN) retention = 1.289 min, [M + H] ⁺ = 6 77.2 Example 1,1 '- (2,3-difluorobutane- Way ^X H NMR (400 lx-ms

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171 1,4-diyl) bis (2- (1-ethyl-Zmethyl-1Н-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-5carboxamide) О ° ⁿ -n hn — I | l ² \ NV / ii g ja ^nh n, ... γ / 8 Ο 15 MHz, DMSO-dg) δ ppm 12.95 (s, 2), 8.03 (s, 2 Η), 8.00 (br. S, 2 Η), 7.82 (d, J = 8, 4 Hz, 2), 7.59 (d, J = 8.4 Hz, 2), 7.36 (bs, 2 Η), 6.67 (s, 2 H), 4.80- 4.92 (m, 2), 4.59 (q, J = 7.2 Hz, 4), 2.06 (s, 6 Η), 1.33 (t, J = 7.1 Hz, 6 N) Method A: retention time NLI = 1.268 min, [M + H] ⁺ = 7 15.2 Example 172 N, N'- (4-carbamoyl8,9,16,17,18,19hexahydro-7Н-6,10-dioxa2,14,15а, 19atetraazacyclopentadeca [1, 2,3-cd: 11,10,9с'd ' ] diindene-1,15diyl) bis (1-ethyl-3-methyl1Н-pyrazole-5-carboxamide ditrifluoroacetate nh ₂ ^{ο; ί} > - / = \ Гд ~ ° ⁰ Г Ν --- N / Ν NH NH Ϋ ν Way 21 ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.6213.07 (m, 1), 8.02 (br.s, 1 Η), 7.67 (s, 1 Η), 7.45 (ο, 1 Η), 7.38 (bs, 1 Η), 7,137.21 (μ, 2 Η), 6.9 6 (dd, 0 = 6.24, 3.06 Hz, 1 Η) , 6.59 (d, 0 = 10.51 Hz, 2), 4.54-4.67 (μ, 6), 4.32-4.53 (μ, 10), 2.30- lx-ms method D: retention time = 1.05 min, [M + H] ⁺ = 7 08.9

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2.36 (m, 1 Η), 2.14-2.25 (m, 1 Η), 2.08-2.14 (m, 6 Η), 2.05 (br. S, 3 Η), 1.26-1.39 (m, 6 N) Example 173 1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -1H-benzo [d] imidazol-1-yl) butyl) -2- (1-ethyl-Zmethyl- 1Н-pyrazole-5carboxamido) -4-hydroxy1Н-benzo [d] imidazole-5carboxamide 2,2,2trifluoroacetate \ __ _{_} o ° PU Ν Ύ Ν '' ΝΗ, ^ν -ν ην-I | Τ ² Η Τ ^ΝΗ а. . ^Η2Ν γΑΑ-Ν ^ О ОН ^{0 44} Method 9 ^X H NMR (400 MHz, CD ₃ OD) δ ppm 7.88 (d, 0 = 1.2 Hz, 1H), 7.77 (dd, 0 = 8.4, 1.2 Hz, 1H), 7 , 65 (d, 0 = 8.8 Hz, 1H), 7.41 (d, 0 = 8.4 Hz, 1H), 6.91 (d, 0 = 8.4 Hz, 1H), 6.59 (d, 0 = 3.2 Hz, 2H), 4.684.59 (m, 4H), 4.29-4.26 (m, 4H), 2.22 (s, 3H), 2.21 (s, 3H), 2.05 (s, 4H), 1.421.38 (m, 6H) LC-MS Method A retention time = 1.36 2 min, [M + H] ⁺ = 6 95.6 Example 174 1,1 '- (ethane-1,2-diyl) bis (2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-5carboxamide) Way 15 ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.53 (s, 2H), 8.01 (s, 2H), 7.9 (s, 2H), 7.89 (s, 2H), 7 , 59 (s, 2H), 7.35 (s, 2H), 7.25 (s, 1H), 7.12 (s, 1H), 6.99 LC-MS Method A retention time = 1.25 min, [M + H] ⁺ = 6 51.2

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V- \ / ° S Ν ΗΝ— {] | Ι ² Π Γ)> - ^{ΝΗ ν} -μ \ / Ν 8 ° ^ Χ (s, 1H), 5.34 (s, 2H), 4.43 (q, 3 = 6, 7 Hz, 4H), 1.95 (s, 6H), 1.24 (t, 3 = 8.0 Hz, 6H) Example 175 1- (2- (Ν- (2- (5-carbamoyl-2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) ΙΗ-benzo [d] imidazole-1yl) ethyl) -2-hydroxyacetamido) ethyl ) -2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-5carboxamide nh ₂ 0+. C /, ^OH N ^ Z / \ r - N ^? A Α ^ν χ A ° G γ '' --NL ο ^ ^ΝΗ \ ___ ⁿ y% X A h ₂ n Way 16 ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.86 (s, 1H), 12.77 (s, 1H), 7.98 (d, 3 = 8.0 Hz, 3H), 7.76 (d, 3 = 8.0 Hz, 2H), 7.517.54-7.48 (m, 2H), 7.35-7.32 (s, 2H), 6.64 (s, 1H), 6, 59 (s, 1H), 4.594.51 (m, 4H), 4.41-4.38 (m, 4H), 3.854.82 (m, 2H), 3.70-3.64 (m, 4H) , 2.092.08 (m, 5H), 1.33-1.30 (m, 5H) LC-MS Method A retention time = 1.23 min, [M + H] ⁺ = 7 51.9 Example 176 1- (2- (2-amino-P- (2- (5carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Нbenzo [d] imidazole-1- yl) ethyl) acetamido) ethyl) -2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-5- Way 15 ^X H NMR (400 MHz, DMSO- ₆ ) δ ppm 12.91 (s, 2H), 8.03-7.99 (m, 6H), 7.817.78 (m, 2H), 7.63 (d , 3 = 8.0 Hz, LC-MS Method A retention time = 1.16 min, [M + H] ⁺ = 7 50.7

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carboxamide nh ₂ “6 · С -г ^ ^{= 0} / - Ο ^ ΝΗ \ 1 η \ ην- ^ Λ-, and ^{0 q} 4 η ₂ ν 1H), 7.55 (d, J = 8.0 Hz, 1H), 7.37 (d, J = 4.0 Hz, 2H), 6.67 (s, 1H), 6.58 (s, 1H), 4.604.50 (m, 6H), 4.42 (d, J = 6.0 Hz, 2H), 3.95 (t, J = 6.0 Hz, 2H), 3.95 (t, J = 6.0 Hz, 2H), 3.56 (d, J = 4.0 Hz, 2H), 2.06 (d, J = 8.0 Hz, 5H), 1.341.30 (m, 5Η) Example 177 8-ethyl-10, 18-dimethyl-7.20 dioxo 6,7,8,11,12,13,14,15,20,21,28,29-dodecahydrobenzo [4,5] imide zo [2,1 - b] benzo [4,5] imidazo [1,2f] dipyrazolo [5,1-j: 4 ', 3' - q] [1,3,6,8,11] pentaazacyclic ononadecin-3,24 dicarboxamide νη ₂ η ₂ ν NH ΗΝ \ ss ° J '' Ν ' ^s \ /] N— \ \ S = NL +, \ / N Me Me' Me ^x Way 19 D NMR (400 MHz, DMSO-dg) δ ppm 12.74 (s, 2Η), 8.06-7.90 (m, 4Η), 7.61 (s, 2Η), 7.34 (s, 2Η ), 6.90 (s, 2Η), 6.28 (s, 1Η), 4.69 (s, 4Η), 4.56 (s, 2Η), 4.27 (s, 2Η), 2.69 (d, J = 12.7 Hz, 2Η), 2.11 (d, J = 22.2 Hz, 6Η), 1.87 (s, 2H), 1.58 (s, 2H), 1.34 (s, 2H), 1.21 (dd, J = 15.2, 8.2 Hz, 3H) LC-MS Method A Retention Time = 1.2 9 9 min, [M + H] ⁺ = 6 91.2

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Example 178 3,24-dicarbamoyl-8-ethyl10,18-dimethyl-7,20-dioxo8,11,12,13,14,15,20,21,28,29,31,32-dodesahydro-1Нbenzo [4,5] imidazo [2,1-b] benzo [4,5] imidazo [1,2i] dipyrazolo [5,1-t: 4 ', 3't] [1,3,6,9,11,14] hexaazac iklodocosin -30-carboxylate h ₂ n nh ₂ y = O X <1 / - N \ \ Boc NH ° <sZ ^NH 1 \ 1> = Ν \ = n / Way 19 ^X H NMR (400 MHz, CD ₃ OD) δ ppm 8.01-7.95 (m, 2Η), 7.86-7.80 (μ, 2Η), 7.507.36 (μ, 2Η), 6, 72-6.59 (μ, 1Η), 4.76 (τ, J = 6 Hz, 1Η), 4.65 (τ, J = 6Hz, 1Η), 4.45-4.42 (m, 4Η) , 4.32 (s, 1Η), 3.783.65 (μ, 4Η), 2.84 (d, J = 8 Hz, 2Η), 2.71 (d, J = 8Hz, 2H), 2.27 ( s, 1H), 2.722.20 (m, 2Η), 2.16 (s, 1Η), 1.88-1.78 (μ, 2Η), 1.521.43 (μ, 2Η), 1.37-1 , 32 (μ, 3Η), 1.231.16 (μ, 2Η), 0.80 (s, 4Η), 0.72 (s, ZN) LC-MS Method A Retention Time = 1.35 6 min, [M + H] ⁺ = 8 34.7 Example 179 8-ethyl-Yu, 18-dimethyl-7,20 dioxo 7,8,11,12,13,14,15,20,21,2 8.29,30,31,32-tetrahydro1H-benzo [4,5] imidazo [2,1b] benzo [4,5] imidazo [1,2i] dipyrazolo [5,1-t: 4 ', 3't] [1,3,6,9,11,14] hexaazats iklodocosin-3,24 dicarboxamide trifluoroacetate Way 19 ^Χ Η NMR (400 MHz, CDO ₃ D) δ ppm 8.06-8.06 (μ, 1Η), 8.01 (s, 1Η), 7.95-7.90 (μ, 2Η), 7, 68 (d, J = 8.0 Hz, 1Η), 7.60 (d, J = 12 Hz, 1H), 6.85 (s, 1Η), LC-MS Method A Retention Time = 1.23 min, [M + H] ⁺ = 7 34.0

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Way

One of two possible structures:

dioxo28,29,30,31idazo [1.21] [1,3,6,15,17]

8-ethyl-Yu, 18-dimethyl-7.20dioxo6,7,8,11,12,13,14,15,20,21

28,29,30,31 tetradecahydrobenzo [4,5] im idazo [1,2а] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'1] [1,3 , 6,15,17] pentaazacycloneicosin-3carboxamide

The structure is not defined.

8-ethyl-Yu, 18-dimethyl-7,206,7,8,11,12,13,14,15,20,21 tetradecahydrobenzo [4.5] im

a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'pentaazacyclogeneicosine24-carboxamide

1.85-1.76 (m 2H), 1.36 (t J = 6.0 Hz, 2H), 1.29 (t J = 6.0 Hz, 2H), 1.19 -

1.12 (m, 2H)

^X H NMR (400 LC-MS MHz, DMSO-d ₆ ) Method A: δ ppm 8.03 time (s, ZN), 7.84 holding (d, 0 = 8.0 Hz, nia = 1.4 8 1H), 7.66 (d, 9 minutes, 0 = 8.0 Hz, [M + H] ⁺ = 6 1H), 7.57- 76, 9 7.55 (s, 1H), 7.37 (s, 1H),

7.28-7.22 (m, 2H), 7.13 (s, 1H), 7.00 (s, 1H), 6.58 (s, 1H), 5.84 (s, 1H), 4 , 75 (t, 2H), 4.47 (q, 0 = 8.0

Hz, 2H),

4.28-4.22 (m,

4H), 2.82 (t,

0 = 8.0 Hz,

2H), 2.68 - 2.66 (m, 1H) 2.34 -2.33 (m 1H), 2.16 (FROM 2H), 2.09 (FROM, 2H), 1.91 (FROM 2H), 1.81 (FROM 2H), 1.51 - 1.47 (m, 4H) 1.29 (t, 0 = 8, 0 Hz, 2H), 1.23 (from 2H)

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h ₂ n m \ ^ N O ^ NH _H / Jr -> ° A ^N JL Z ^N - \ / Z% ^? \ Example 182 The structure is not defined. One of two possible structures: 8-ethyl-10, 18-dimethyl-7,20dioxo-6,7,8,11,12,13,14,15,20,21, 28,29,30,31-tetradecahydrobenzo [ 4,5] im idazo [1,2-a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3'1] [1,3,6,15,17 ] pentaazacyclogeneycosine24-carboxamide nh ₂ Ό К? m NH HN ^Οί Ζ yY Ν \ / 1 ^Ν -V \ = ^N An ' ^X 8-ethyl-Y, 18-dimethyl-7.20 dioxo-6,7,8,11,12,13,14,15,20 , 21, 28,29,30,31-tetradecahydrobenzo [4,5] im idazo [1,2-a] benzo [4,5] imidazo [2,1p] dipyrazolo [5,1-e: 4 ', 3 '1] [1,3,6,15,17] pentaazacycloneicosine-3carboxamide Way 19 ^X H NMR (400 MHz, DMSO-s1 ₆ ) δ ppm 8.037.99 (m, 2H), 7.84-7.80 (m, 1H), 7.627.54 (m, 3H), 7.38-7 , 35 (m, 1H), 7.617.54 (m, 2H), 7.10 (s, 1H), 6.97 (s, 1H), 6.57-6.55 (m, 1H), 4.774, 72 (m, 2H), 4.50-4.45 (m, 2H), 4.274.22 (m, 3H), 2.85-2.80 (m, 2H), 2.692.67 (m, 1H) , 2.34-2.32 (m, 1H), 2.172.13 (m, ZN), 2.10-2.07 (m, ZN), 1.931.89 (m, ZN), 1.83-1 , 79 (m, 2H), 1.511.46 (m, 2H), 1.40-1.35 (m, 2H), 1.29 (t, J = 6.0 Hz, 2H), 1.241.22 ( m, 1H) LC-MS Method A Retention Time = 1.4 8 9 min, [M + H] ⁺ = 6 77.0

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h ₂ n Cha U ^Ν γ K ^ N O ^ ^NH HN X -—> ° L D / A Example 183 10,18-dimethyl-7,20-dioxo6,7,8,11,12,13,14, - 15,20,21,28,29,30,31 tetradecahydrobenzo [4,5] im idazo [1,2- a] benzo [4,5] imidazo [2,1-p] dipyrazolo [5,1-e: 4 ', 3'- 1] [1,3,6,15,17] pentaazacyclogeneicosine-3,24-dicarboxamide NH ₂ H ₂ N _p b i N 1 ._____ J ^,, Ν - ^ΗΝ ΗΝ V = N y == N Way 19 ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.91 (s, 1H), 12.76 (s, 1H), 8.03 (s, 4H), 7.83 (m, 2H), 7.697 , 62 (m, 2H), 7.37 (br s, 2H), 6.57 (s, 1H), 4.75 (br s, 2H), 4.26 (br s, 4H), 2.87 (br s, 2H), 2.16-1.24 (m, 17H) LC-MS Method A retention time = 1.2 8 3 min, [M + H] ⁺ = 6 91.2 Example 184 (Ε) -7- (aminomethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -7-methoxy1H-benzo [d] imidazol-1yl) but-2-en-1-yl) -2- (1ethyl-3 -methyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide Method 9 ^X H NMR (400 MHz, DMSO-dg) δ ppm 8.15 (s, 1H), 7.95 (t, J = 14.8 Hz, 3H), 7.68 (d, T = 25.8 Hz , 2H), 7.35 (D, J = 18, l Hz, 3H), 6.52 (d, J = 18.5 Hz, 2H), 5.76 (D, J = 15.9 Hz, 1H ), 5.54 (d, J = 15.8 LC-MS Method A Retention Time = 1.2 0 5 min, [M + H] ⁺ = 7 35.8

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N / —A / \ - / ° ° = \, ^Ν '^ ^Ζ ^ Γ ^ ΝΗ ₂ hn— {jj J Ν' ^ γ О ^Н ₂ М // Г ΪΓ Ζ> ~ ^ΝΗ Ν ^ .. η ₂ νy -ς JJ 8 θ Hz, 1H), 5.08 (s, 2H), 4.88 (s, 2H), 4.51 (d, 0 = 6.7 Hz, 4H), 3.95 (s, 2H), 3, 76 (s, 3H), 2.10 (d, 0 = 1.8 Hz, 6H), 1.26 (td, 0 = 7.1, 4.5 Hz, 6H) Example 185 (Ε) -7- (aminomethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -7- (3hydroxypropoxy) -1Hbenzo [d] imidazole-1- yl) but2-en-1-yl) -2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide D,? Г // LA ΗΝ — σ || 1 NH ₂ °> h (1 h ₂ n L £ A _x ^h _z ⁿ -n ^oh 8 ° Method 9 ^X H NMR (400 MHz, DMSO-dg) δ ppm 8.15 (s, 1H), 7.95 (t, 0 = 17.8 Hz, 3H), 7.70 (s, 1H), 7.64 (s, 1H), 7.34 (m, 3H), 6.52 (d, 0 = 9.9 Hz, 2H), 5.78 (d, 0 = 15.9 Hz, 1H), 5.56 (d, 0 = 15.8 Hz, 1H), 5.09 (s, 2H), 4.90 (s, 2H), 4.52 (d, 0 = 6.6 Hz, 4H), 4.04 (s, 2H), 3.91 (s, 2H), 3.41 (s, 2H), 2.10 (s, 6H), 1.62 (m, 2H), 1.27 (t, 0 = 7.1 Hz, 6H) LC-MS Method A Retention Time = 1.18 6 min, [M + H] ⁺ = 7 79.8 Example 186 1,1 '- (butane-1,4- diyl) bis (2- (1-allyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-5carboxamide) Way eleven ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.81 (s, 2H), 7.97 (s, 4H), 7.76 (d, 0 = 8.34 lx-ms method C: time retention = 0.83 min,

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V \ ° jj nI '// ^NH 2 ⁿ n HN-1 Π nA / ¹ , ζ ^ ν-Ν Η Τ Α ^{ΝΗ Ν} -μ Η ₂ Ν \ / Ν 8 θ Hz, 2H), 7.53 (d, J = 8.34 Hz, 2 Η), 7.33 (br s, 2H), 6.64 (s, 2H), 5.936, 05 (m, 2H), 5.21 (d, J = 4.80 Hz, 4 Η ), 5.02 (dd, J = 10.36, 1.01 Hz, 2H), 4.89 (dd, J = 17.18, 1.01 Hz, 2H), 4.27 (br s, 4H), 2.10 (s, 6H), 1.86 (br s, 4H) [M + H] ⁺ = 7 03.6 Example 187 1,1 '- (butane-1,4-diyl) bis (2- (1-ETHYL-4-IOD3-methyl-1H-pyrazole-5carboxamido) -1H-benzo [d] imidazole-5carboxamide) 1 Me. / o ° 11 7 ~ X NZ '/ # ιΤ ΝΗο Ν ΗΝ - V 1 ² ) Me <) Me iTW (^ γΜ-Lul o ° / ^ Me 1 Way eleven ^T H NMR (400 MHz, DMSO- ₆ ) δ ppm 12.92 (s, 2H), 8.00 (D, J = 1, 01 Hz, 2H), 7.98 (br. S, 2H), 7.78 (dd, J = 8.46, 1.39 Hz, 2H), 7.57 (d, l7 = 8.34 Hz, 2H), 7.35 (br s, 2 Η), 4, 52 (q, J = 7.07 Hz, 4 Η), 4.35 (br s, 4H), 2.14 (s, 6H), 1.91 (br s, 4H), 1.28 (t, J = 7.07 Hz, 6H) LC-MS Method C Retention Time = 1.01 min, [M + H] ⁺ = 9 31.5 Example 1-allyl-2- (1- (5- (5 - ((1- Method 7 ^X H NMR (400 LC-MS

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188 allyl-5-carbamoyl-1Hbenzo [d] imidazole-2yl) carbamoyl) -1-ethyl-Zmethyl-1H-pyrazole-4yl) pentyl) -3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole- 5carboxamide [/ Me / O G> - / NI II ^h 2 ⁿ ^ AA-i / P < ^{Me 0} \ ⁰ / n n ~ -n ί) MHz, DMSO-dg) δ ppm 12.88 (s, 1H), 12.81 (s, 1 Η), 7.99-8.02 (m, 2H), 7.97 (br.s ,, 2 Η), 7.77 (ddd, 0 = 8.34, 3.66, 1.39 Hz, 2), 7.41 (dd, 0 = 16.93, 8.34 Hz, 2), 7 , 34 (br s, 2), 6, 65 (s, 1H), 5.87-6.02 (m, 2 Η), 4.995.22 (m, 4H), 4.82 (dd, 0 = 11.62, 4.80 Hz, 4H), 4.50-4.61 (m, 4H), 2.73 (t, 0 = 7.45 Hz, 2H), 2.15 (s, 3H) , 2.08 (s, 3H), 1.711.85 (m, 2H), 1.45-1.55 (m, 2H), 1.271.34 (m, 5H) method C: retention time = 0.93 min, [M + H] ⁺ = 7 45.7 Example 189 8-ethyl-Yu, 18-dimethyl-7,20 dioxo 6,7,8,11,12,13,14,15,20,21,28,31- dodecahydrobenzo [4,5] imide zo [1,2- a] benzo [4,5] imidazo [2,1- p] dipyrazolo [5,1 - e: 4 ', 3' - 1] [1,3,6,15,17] Method 7 Data are given for trans-isomer ^X H NMR (400 MHz, DMSO-dg) δ ppm 12.87 (s, 1H), 12.84 (s, 1H), 7.98 Data shown for trans isomer LC-MS Method C: retention time

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Example

190

2- (1- (5- (5- ((5-carbamoyl- Method

1-propyl-1H-22 benzo [d] imidazole-2yl) carbamoyl) -1-ethyl-Zmethyl-1H-pyrazole-4yl) pentyl) -3-methyl-1H-pyrazole-5-carboxamido) -1propyl-1Hbenzo [d] imidazole-5carboxamide

(br. s, 4H), nia = 0.82 7.77 (dd, MIN, J = 7.71, 3.16 [M + H] ⁺ = 7 Hz, 2H), 7.33-7.48 (m, 4H), 6.55 (s, 1 Η), 5.895.98 (m, 1 Η), 5.66-5.75 (m, 1 Η ), 4.90 (d, J = 7.83 Hz, 4 Η), 4.73 (t, J = 6.95 Hz, 2H), 4.47 (q, J = 6.99 Hz, 2H) , 2.72-2.80 (m, 2H), 2.17 (s, 3H), 2.10 (s, 3H), 1.72 (br s, 2H), 1.44 (br s , 2H), 1.30 (t, J = 7.07 Hz, 5H) 17.6 ^X H NMR (400 LC-MS MHz, DMSO-d ₆ ) method C: δ ppm 12.84 time (s, 1H), holding 12.78 (s, 1 nia = 0.97 H), 8.00 (d, MIN, J = 2.21 Hz, 2 [M + H] ⁺ = 7 Η), 7.97 (br s, 2 H), 7.78 (d, J = 7.07 Hz, 2 Η), 7.49-7.58 (m, 2 Η), 7.33 ( width s, 2 Η), 6.63 (s, 1 Η), 49.7

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Me S о Г _ ki W Π Г / А / \\ I / ^h 2 ⁿ VYA-n ^ П / Me ⁰ \ ⁰ / ^H 2 ^N ^ Ti ^X % ^ '\ H NN | ly ~ ^N x_Z \\ \ F-M? ⁰ Me 4.51-4.65 (m, 4), 4.074.20 (m, 4), 2.76 (t, 0 = 7.07 Hz, 2 Η), 2.15 (s, 3) , 2.08 (s, 3 Η), 1.69-1.83 (m, 6 Η), 1.481.59 (m, 2 Η), 1.28-1.36 (m, 5H), 0.83-0.91 (m, 6H) Example 191 ethyl 4- (2- (1- (5- (5 - ((1allyl-5-carbamoyl-1H-benzo [d] imidazol-2- yl) carbamoyl) -1-ethyl-3-methyl-1H-pyrazole-4- yl) pentyl) -3-methyl-1Hpyrazole-5-carboxamido) -5carbamoyl-1H-benzo [d] imidazol-1- yl) butanoate II Me S o <___ \\ n A- / NI /) - ΝΗ Λ / / II / Me О) О / H, N Ж 'Х Η Ν-Ν IL JL / ^Ν νΛ \ к fl' ^^^ Ме 1 ° \ X ° OEt Way 23 ^X H NMR (400 MHz, (CDCl ₃ methanol D ₄ , 1: 1) δ ppm 7.89 (s, 1H), 7.87 (s, 1H), 7.82 (d, 0 = 8.28 Hz, 1H), 7.78 (d, 0 = 8.28 Hz, 1H), 7.37 (d, 0 = 8.53 Hz, 1H), 7.24 (d, 0 = 8.53 Hz, 1H) , 6.68 (s, 1H), 5.835.95 (m, 1H), 5.20 (d, 0 = 10.29 Hz, 1H), 5.07 (d, 0 = 17.32 Hz, 1H ), 4.77 (d, 0 = 4.27 Hz, 2H), 4.594.65 (m, 2H), 4.54 (q, 0 = 6.94 Hz, 2 LC-MS Method D Retention Time = 1.01 min, [M + H] ⁺ = 8 19.7

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Η), 4.23 (τ, J = 6.65 Hz, 2Η), 4.03 (q, J = 7.19 Hz, 2 Η), 2.79 (τ, J = 7.40 Hz, 2Η ), 2.38 (τ, J = 6.90 Hz, 2 Η), 2.23 (s, 3 Η), 2.15 (s, 3 Η), 2.08-2.14 (μ, 2Η ), 1.801.92 (μ, 2Η), 1.51-1.63 (μ, 2), 1.36 (τ, J = 7.03 Hz, 5 Η), 1.15 (t, J = 7.03 Hz, ZN) Example 192 ethyl 4- (5-carbamoyl-2- (1 (5- (5- ((5-carbamoyl-1Нbenzo [d] imidazole-2- yl) carbamoyl) -1-ethyl-3- methyl-1H-pyrazole-4- yl) pentyl) -3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butanoate Me n⁰ ___ P \\ N. Γ /> - ^ \\ II Π \^{Me 0} \ ⁰ / η₂ν ΥΎχΙ η ν ~ ν [II τ Ν fl Me L⁰ OEt Way 23 ^X H NMR (400 MHz, DMSO-dg) δ ppm 7.11 (s, 1H), 7.07 (s, 1H), 6.99 (dd, J = 8.44, 1.34 Hz, 1H) , 6.91 (dd, J = 8.31, 1.47 Hz, 1H), 6.58 (D, J = 8.5 6 Hz, 1H), 6.54 (D, J = 8.5 6 Hz, 1H), 5.78 (s, 1H), 4.04 (br s, 5H), 3.75 (t, J = 6.85 Hz, 2H), 3.54 (q, J = 6 , 85 LC-MS Method D Retention Time = 0.8 8 min, [M + H] ⁺ = 7 79.6

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Hz, 2H), 3.37 (t, 0 = 6.72 Hz, 2H), 3.14 (q, 0 = 7.09 Hz, 2H), 1.84 (t, 0 = 7.09 Hz, 2 Η), 1.50-1.57 (m, 2H) , 1.33 (s, ZN), 1.32 (s, ZN), 1.191.28 (m, 2 Η), 0.92-1.02 (m, 2 Η), 0.64-0.73 (m, 2H), 0.52 (t, 0 = 7.09 Hz, ZN), 0.400.49 ( m, 2 Η), 0.28 (t, 0 = 7.21 Hz, ZN) Example 193 3- (2- (1- (5- (5- ((1- (2- (((benzyloxy) carbonyl) amino) ethyl) -5-carbamoyl-1Hbenzo [d] imidazole-2- yl) carbamoyl) -1-ethyl-Zmethyl-1H-pyrazole-4- yl) pentyl) -3-methyl-1Hpyrazole-5-carboxamido) -5carbamoyl-1H- benzo [d] imidazole-1yl) propanoic acid Way 23 ^X H NMR (400 MHz, (methanol-b ₄ ) δ 7.91 (s, 1H), 7.77-7.82 (m, 2H), 7.70 (dd, 0 = 8.56, 1, 47 Hz, 1 H), 7.44 (d, 0 = 8.31 Hz, 1H), 7.33 (d, 0 = 8.56 Hz, 1H), 7.127.30 (m, 5H), 6, 69 (s, 1 Η), 4.93 (s, 2 Η), 4.494.65 (m, 4H), 4.41 (t, 0 = 6.85 Hz, LC-MS Method E Retention Time = 0.81 min, [M + H] ⁺ = 9 14.5

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NHCbz 1 Me 1 o \ Η Τ / Ay ^N / A] G ^N / Me O / CO ₂ N / Η Ν-Ν || I> - ^Ν ^ ___ Ζ \\ ^ Α ^ ζΆ _Με P o o 2H), 4.30 (τ, J = 5.75 Hz, 2Η), 3.55 (τ, J = 5.75 Hz, 2Η), 2.88 (τ, J = 6.72 Hz, 2 Η ), 2.75-2.82 (μ, 2Η), 2.21 (s, ZN), 2.16 (s, ZN), 1.81-1.91 (m, 2Η), 1.531.63 (μ, 2Η), 1.36 (τ, J = 7.09 Hz, 5Η) Example 194 methyl 1- (4- (5-carbamoyl-2 (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-1yl) butyl) -2- (1-ethyl-Zmethyl-1H -pyrazole-5carboxamido) -7-methoxy1Н-benzo [d] imidazole-5carboxylate А | V А A 7 - nh ₂ ° Ι ^ ΐΓ ψχ W ^Ν == {' _Ν NH J О = / θ ^ ΝΗ # ^{N |} rA ^N ^ \ 1 Way eleven ^Χ Η NMR (400 MHz, DMSO-dg) δ ppm 12.6613.03 (μ, 2Η) 7.97 (s, 2Η) 7.78 (s, 2Η) 7.50-7.57 (m, 1Η ) 7.37 (d, J = l, 01 Hz, 2 Η) 6.59 (s, 2 Η) 4.57 (br s, 4H) 4.344.44 (m, 2H) 4.20-4, 31 (m, 2H) 3.88 (d, J = 3.79 Hz, 6H) 2.11 (s, 6H) 1.86 (br s, 4H) 1.27-1.37 (m, 6H ) LC-MS Method C Retention Time = 0.93 min, [M + H] ⁺ = 7 24.6 Example 195 2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) -1 (4- (2- (l-ethyl-3-methyl-1Hpyrazole-5-carboxamido) ΙΗ-benzo [d] imidazole-1- Way eleven ^X H NMR (400 MHz, DMSO-dg) δ ppm 1.25 (t, J = 6.95 Hz, 6H) 1.76 LC-MS method C: time holding nia = 0.8 8

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yl) butyl) -1Нbenzo [d] imidazole-5carboxamide Och / X 'n 1 - JI ν and m D JL <7 ”'ν ABOUT ' (br s, 4H) 2.10 (s, 6H) 4.11 (br s, 4H) 4.50-4.68 (m, 4H) 6.35 (s, 2H) 6.847.03 (m , 4H) 7.12 (d, J = 7.07 Hz, 2 H) 7.36-7.52 (m, 2H) 7.68 (br s, 1H) 8.00 (s, 1H) min, [M + H] + = 6 36, 5 Example 196 tert-butyl bis (2- (5carbamoyl-2- (1-ethyl-Zmethyl-1H-pyrazole-5carboxamido) -1Hbenzo [d] imidazole-1yl) ethyl) carbamate 0 7 ~ ^NH 2> ADR Ν | H n> 0 ... V 1 JL А о о ζ Way 15 ^X H NMR (400 MHz, (methanol-b ₄ ) δ 0.96-1.17 (m, 9H) 1.25-1.49 (m, 6H) 2.082.37 (m, 6 H) 3.58 (br s, 4H) 4.05-4.39 (m, 4H) 4.62 (br s, 4H) 6.38-6.63 (m, 2H) 7.06-7.30 (m , 2H) 7.63 (br s, 2H) 7.96 (br s, 2H) LC-MS Method C: retention time = 0.83 min, [M + H] + = 7 94.7 Example 197 7,7 '- (propane-1,3diylbis (oxy)) bis (1-allyl2- (1-ethyl-3-methyl-1Hpyrazole-5-carboxamido) 1H-benzo [d] imidazole-5carboxamide) ditrifluoroacetate Way 21 ^X H NMR (400 MHz, DMSO-dg) δ ppm 1.34 (t, J = 7.07 Hz, 6H) 2.18 (s, 6H) 2.332.46 (m, 2H) 4.41 (t, J = 5.81 Hz, 4 H) 4.60 (q, J = 7.07 Hz, 4H) 4.85-5.11 LC-MS Method C: retention time = 0.94 min. [M + H] + = 7 77.7

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Ligand for analysis AlexaFluor-488 FRET ', 6'-Diamino-5 - ((2- (1 - (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido ) -1H-benzo ^] imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-III-pyrazole-5-carboxamido) -1H-benzo ^] imidazole-5carboxamido) ethyl) carbamoyl) -3- oxo-3H-spiro [isobenzofuran-1,9'-xanthene] -4 ', 5'-disulfonic acid

- 218

- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo-imidazol-1 yl) buty.l) -2- ( '1-ethyl.l-3-methyl.l-1H-pyrazo.l-5-carboxamido) -1H-benzoLimidazo.l-5-carboxylic acid dihydrochloride

K methyl-1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-1yl) butyl) -2- (1-ethyl -3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5-carboxylate bistrifluoroacetate (400 mg, 0.434 mmol, example 23) in THF (3.47 ml), MeOH (3.47 ml) and water (1.74 ml) at room temperature was added 8M potassium hydroxide solution (1.09 ml, 8.68 mmol). After stirring overnight, the reaction mixture was concentrated and water was added. The mixture was acidified with 7N HCl aqueous solution to pH 4-5, and the resulting gray solid was collected by filtration to give the title compound (335 mg, 0.423 mmol, 97% yield). ^{1 * 3} H NMR (400 MHz, DMSOd6) δ ppm 12.82-12.95 (m, 3H), 8.08 (s, 1H), 7.99 (br s, 2H), 7.83 ( d, J = 8.34 Hz, 1H), 7.78 (d, J = 8.34 Hz, 1H), 7.58 (t, J = 7.33 Hz, 2H), 7.36 (br. s, 1H), 6.60 (d, J = 4.80 Hz, 2 H), 4.58 (d, J = 6.57 Hz, 4H), 4.29 (br s, 4H,) 2 , 10 (s, 6H), 1.88 (br s, 4H), 1.31 (t, J = 6.95 Hz, 6H); LCMS (LCMS Method C): retention time = 0.83 min, [M + H] + = 680.5.

Stage 1:

C- (2-aminoethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazol-1-yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5carboxamide trifluoroacetate

- (4- (5-Carbamoyl-2- (1-ethyl-3-methyl-1 H-pyrazole-5-carboxamido) -1 H-benzo ^ imidazol-1 yl) butyl) -2- (1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5-carboxylic acid (10 mg, 0.015 mmol) was dissolved (sonicated) in DMSO (300 μl) at 37 ° C. To this was added a solution of (9H-fluoren-9-yl) methyl (2-aminoethyl) carbamate hydrochloride (6.9 mg, 0.022 mmol) and HATU (7.6 mg, 0.020 mmol) in DMSO (100 μl), and then DIEA (10 μl, 0.057 mmol). After stirring overnight, the reaction mixture was diluted with DMF (600 μl), 4-methylpiperidine (400 μl) was added and the reaction mixture was stirred at room temperature for 1 h. The mixture was concentrated and the resulting residue was diluted 1: 1 with DMSO: MeOH (<1 ml) and purified by reverse phase chromatography (Jupiter C18 preparative column, 10 ml / min) eluting with 30-100% (9: 1 ACN: water) in water (0.1% TFA added) to afford the title compound ( 8.45 mg, 10.1 μmol, 69% yield). LCMS (LCMS Method G): retention time = 0.62 min, [M + H] + = 722.4.

Stage 2:

', 6'-Diamino-5 - ((2- (1 - (4- (5 -carbamoyl-2- (1-ethyl-3-methyl- 1H-pyrazole-5-carboxamido) -1НbenzoU] imidazole-1- yl) butyl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzoU] imidazole-5carboxamido) ethyl) carbamoyl) -3-oxo-3H-spiro [isobenzofuran-1, 9'-xanthene] -4 ', 5'-disulfonic acid

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k- (2-Aminoethyl) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1Hbenzo ^] imidazol-1-yl) butyl) - 2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -1H-benzo Schimidazole-5carboxamide trifluoroacetate (8.45 mg, 10.1 μmol) was dissolved in DMF (200 μL) and added to solid ( 5,6-) Alexa Fluor 488-ONSu (5.00 mg, 7.92 μmol). The commercial reagent Alexa Fluor 488-ONSu was a mixture of 5- and 6-position isomers.

When the solution was prepared, DIPEA (2 μl, 0.01 mmol) was added and the mixture was vortexed overnight in the dark. The LC-MS results showed the appearance of peaks of early and late elution products with an assumed molecular weight ([M + H] 1238.6). The reaction mixture was concentrated and the residue was dissolved in 1: 1 DMSO: MeOH (<1 ml) and purified by reverse phase chromatography (Jupiter C18 preparative column, 10 ml / min) eluting with 15-100% (9: 1 ACN: water ) in water (with the addition of 0.1% TPA). The previously eluted positional isomer was obtained with high purity. In contrast, the fractions of the later eluted isomer also contained unreacted starting material. These fractions, containing the unclean late eluted isomer, were pooled and concentrated. The resulting residue was dissolved in 1: 1 DMSO: MeOH (<1 ml) and purified by reverse phase chromatography (Waters SymmetryPrep preparative column, 10 ml / min) eluting with 15-100% (9: 1 ACN: water) in water (c adding 0.1% TFA) to afford the title compound (late eluted isomer, 1.94 mg, 1.49 μmol, 19% yield). LCMS (LCMS Method H): retention time = 0.69 min, [M + H] ⁺ = 1238.6.

It should be noted that the proposed structure of the title compound (5-isomer) is not based on rigorous structural determination, but is based on previous findings that the 5-position isomer is usually the later eluting isomer in reverse phase HPLC.

Biological tests and data

As indicated above, the compounds of the present invention are STING modulators and are useful in the treatment of STING-mediated diseases. The biological activity of the compounds of the present invention can be determined using any suitable assay to determine the activity of the compound as a STING modulator, as well as in vivo models and tissues.

The pIC ₅₀ value for each compound was obtained in at least one experiment, or an average of several experiments. It is understood that the data described herein may vary reasonably depending on the specific conditions and procedures used by the experimenter.

Binding Analysis (1) SPA

A radioligand binding assay was developed to measure quantitative interactions between a compound of the invention and the carboxy terminal domain (CTD) STING by competing with 3H-cGAMP (tritiated cyclic guanine- (2 ', 5') - monophosphate-adenine- (3 ', 5') -monophosphate). See also Li et

- 220 036435 al. (Nature Chemical Biology, 10, 1043-1048, (2014)). The sequence coding for residues ranging from 149 to 379 of the human STING protein (gene ID 340061) was expressed in bacteria with a Flag® carboxyterminal peptide fused to AviTag ™ for biotinylation and a hexahistidine tag to provide affinity purification. The purified STING-Flag-AviTag-6Xhis protein was biotinylated to completion using the BirA enzyme (Beckett D. et al., Protein Science, 1999, 8: 921-929). The relative activity of a compound of the invention was determined by competing in equilibrium binding reactions containing 50 nM biotinylated-STING, 50 nM 3H-cGAMP and 1.25 mg / ml using a streptavidin coated bead scintillation assay (Perkin Elmer) in phosphate buffered saline physiological buffer. Binding reactions were incubated at room temperature for 30 min and analyzed on a fluorescent plate reader. The concentration-response curves were normalized to control, which reflects complete inhibition of 3H-cGAMP binding by 10 mM unlabeled cGAMP, and without any control compound. The apparent pIC _{50 was} determined using a conventional two-state binding model. Under these conditions, the apparent inhibition constant for the positive control compound with GAMP is 40-50 nM, which is about ten times higher than its actual affinity, leaving 4-5 nM (Zhang X. et al., Molecular Cell, 2013, 51: 1- ten).

(2) FRET analysis

The binding activity of the molecules to the C-terminal domain (CTD) of human STING was determined using a competitive binding assay. This assay used a recombinant STING protein (149-379) with a C-terminal biotinylated Avi tag. Upon binding to STING, the probe of the active site Alex488 (see pages 226-229 for ligand synthesis for FRET assay) receives 490 nm emission from Tb-streptavidin-Avi-STING, and the increase in fluorescence is measured at 520 nm. Molecules that compete for the probe binding site will lower the signal at 520 nm. The assay was performed in black 384-well Greiner plates (cat # 784076) containing 100 nl of compounds in pure DMSO. 500 pM STING solution, 500 pM streptavidin-Lumi4-Tb and 100 nM Alix488 probe solution in phosphate buffered saline containing 0.02% (w / v) Pluronic F127 and 0.02% (w / v) ) bovine serum albumin was added to the plate using a Combi liquid dispenser (ThermoFisher). The plates were centrifuged for 1 min at 500 rpm, incubated for 15 min at room temperature, and then the fluorescence emission at 520 nm was measured after laser excitation at 337 nm on an Envision plate reader (Perkin-Elmer). PIC ₅₀ values were determined using a standard four-parameter curve corresponding to ABASE XE.

Using the above SPA analysis, it was found that the compounds of examples 1-9, 11, 13, 15, 16, 23, 25-30, 33-35, 47, 49, 50, 54, 55, 57-61, 63 -66, 68-74, 76-79, 81-83, 85-88, 90, 92, 102, 104, 105, 107-110, 112-114, 117, 118, 120, 122-144, 146-149 , 151-160, 163-167, 169, 170, 172-183 and 186-197 had pIC ₅₀ values ranging from 3.6 to 7.7. For example, the compounds of Example 1 and Example 5 above, inhibited the binding of MN-cGAMP with STING according to the method described above with a mean pIC ₅₀ 7.5 (# 1, n = 4, # 5, n = 2).

Using the above FRET analysis, Examples 1-106, 110, 112-125, 129, 131, 133, 134, 138, 142144, 146-153, 155-186, 188-193 and 196 showed pIC ₅₀ values in the range of 4, 1 up to the upper limit of analysis at 9.9. For example, the pIC ₅₀ values in the FRET assay for the following examples are:

The function of the compounds of formula (I) can be evaluated in cellular tests in which specific activation of STING and / or inhibition of IFNe protein secretion is detected.

(1) Functional Assay I (PBMC Antagonist Assay): STING inhibition by compounds of formula (I-N), (I-P) or (I) can be determined by measuring the loss of interferon β secreted from PBMCs stimulated by Bacmam virus, double-stranded DNA virus, after

221 036435 treatments with various concentrations of a compound of the invention.

Frozen PBMC cells were thawed and diluted in medium (RPMI-1640 with 1.5 g / L NaHCO ₃ , 4.5 g / L glucose, 10 mM Hepes and 1 mM Na pyruvate, 10% FBS) to a final titer of 5x05 cells / ml followed by infection with Bacmam virus at a final MOI of 43. The PBMC-Bacmam virus suspension was dispensed into a 384-well tissue culture plate (Griener 781073) at 25,000 cells per well containing 250 nL of compound dissolved in DMSO. The level of IFN | 3 protein secreted into the culture medium was measured after 24 h incubation at 37 ° C. using a human IFN® electrochemiluminescence kit (Meso Scale Diagnostics) according to the manufacturer's instructions. Percent inhibition was determined relative to controls that were not treated with compound or lacked Bacmam virus and compound concentration was plotted to determine pIC ₅₀ using a standard bipate receptor-ligand inhibition model.

(2) Functional Assay II (PBMC Agonist Assay): STING activation by compounds of formula I was determined by measuring levels of JFNp secreted from human peripheral blood mononuclear cells (PBMCs) treated with various concentrations of compounds of formula I. Frozen PBMCs were thawed in medium (RPMI-1640 with 1.5 g / L NaHCO ₃ , 4.5 g / L glucose, 10 mM Hepes and 1 mM Na pyruvate, 10% FBS, 10 ng / ml lipopolysaccharide) to a final titer of 5x105 cells / ml and dispensed into a 384-well a tissue culture plate (Griener 781073) at the rate of 15,000 cells per well, containing 250 nl of the compound dissolved in DMSO. The level of JFNp protein secreted into the culture medium was measured after 24 h incubation at 37 ° C. using a human IFN® electrochemiluminescence kit (Meso Scale Diagnostics) according to the manufacturer's instructions. Percent activation was determined versus control treatment with DMSO and plotted versus compound concentration to determine pEC ₅₀ using a standard receptor activation model.

(3) Functional Assay III (HEK WT Cell Agonist Assay): STING activation in cells can be determined using a luciferase reporter assay in human embryonic kidney cells (HEK293T) co-transfected with STING-expressing plasmids and the enzyme firefly luciferase regulated by interferon-stimulated the response promoter element (pISRE-Luc) (Agilent Technologies). Full length human STING (Gene ID 340061) and full length cyclic GMP-AMP synthetase (cGAS) (reference sequence NM 138441.2) were cloned into mammalian cell expression vectors containing the cytomegalovirus promoter. Transfections were performed using a cell suspension with Fugene® 6 according to the manufacturer's instructions (3: 1 Fugene®: DNA). Fifty microliters of the transfection suspension were dispensed into the wells of a 384-well plate containing 250 nl of a compound of the invention. The final well composition contained 20,000 cells / well, 1 ng STING, 20 ng pISRE-Luc and pcDNA3.1 empty vector (Invitrogen) to bring the total DNA concentration to 125 ng. Control wells, which, as expected, produced the maximum STING activation, were cotransfected with the cGAS expression plasmid. The plates were sealed and incubated for 24 hours at 37 ° C. Firefly luciferase expression was measured using a Steady-Glo® luciferase assay system (Promega) and analyzed using a conventional luminescent laboratory plate reader. Data were normalized to the luminescence response in the presence of cGAS, plotted against compound concentration, and fitted using a standard receptor activation model to obtain pEC ₅₀ .

Using the above functional assay III (HEK WT cell agonist assay), the compounds of Examples 1-23, 25-42, 44, 47-55, 57-61, 63-94 and 97-197 showed pEC ₅₀ values ranging from 4 , 4 up to the upper limit of analysis 9.1.

Examples 1, 3, 8, 100, 116 and 194 showed a pEC ₅₀ below 4.3. For example, pEC ₅₀ for the following examples are:

Example No. Agonist assay for HEK WT (pEC ₅₀ ) ten 7.5 eleven 7, 8 13 7.4 14 7.5 16 7, 6 eighteen 7.5 19 7.5 21 7.3 27 7, 6 31 7, 8

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Claims (12)

CLAIM 1. A compound which is (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -7- (3-morpholinopropoxy) -1H-benzo ^ ] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzo ^ | imidazole-5-carboxamide having the structure: or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1, which is a pharmaceutically acceptable salt (E) -1- (4- (5carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7- (3- morpholinopropoxy) -1H-benzoU] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido) -7-methoxy-1H- benzoU] imidazole 5-carboxamide. 3. A compound according to claim 1, which is (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -7- (3-morpholinopropoxy) - 1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl 1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzoIimidazole-5- carboxamide. 4. A compound according to claim 1, which is (E) -1- (4- (5-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5carboxamido) -7- (3-morpholinopropoxy) - 1H-benzo ^] imidazol-1-yl) but-2-en-1-yl) -2- (1-ethyl-3-methyl 1H-pyrazole-5-carboxamido) -7-methoxy-1H-benzoIimidazole-5- carboxamide tris hydrochloride. 5. A pharmaceutical composition for the treatment of a STING-mediated disease, comprising a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 and a pharmaceutically acceptable excipient. 6. A method of treating a STING-mediated disease, comprising administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 to a person in need thereof. 7. The method of claim 6, wherein the disease is cancer. 8. The method of claim 6, wherein the disease is a solid tumor. 9. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 in the treatment of a STING-mediated disease. 10. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 in the manufacture of a medicament for the treatment of a STING-mediated disease. 11. Use according to any one of claims 9 or 10, wherein the disease is a solid tumor. 12. Use according to any one of claims 9 or 10, wherein the disease is cancer.