Fused heteroaryl hydroxamates as STING agonists

Abstract

The present invention provides novel substituted heterocyclic compounds represented by formula I, or pharma- ceutically acceptable salts, solvates, polymorphs, esters, tautomers or prodrugs thereof, and compositions comprising these compounds. The provided compounds can be used as agonists of stimulator of interferon genes (STING), and are useful in the treatment of cancer and certain infectious diseases.

Description

[Technical field]

[0001] The present invention relates to a series of compounds that are stimulator of interferon genes (STING) transforming agents, which are useful in treating breast cancer, ovarian cancer, prostate cancer, testicular cancer, urothelial carcinoma, bladder cancer, non-small cell lung cancer, small cell lung cancer, sarcoma, colorectal adenocarcinoma, gastrointestinal stromal tumor, gastroesophageal carcinoma, colon cancer, pancreatic cancer, renal cancer, hepatocellular carcinoma, malignant mesothelioma, leukemia, lymphoma, myelodysplastic syndrome, multiple myeloma, transitional cell carcinoma, neuroblastoma, plasma cell neoplasms, Wilms' tumor, and other cancers. tumor), hepatocellular carcinoma, urinary tract cancer, bladder carcinoma, colorectal cancer, colon cancer, breast cancer, prostate cancer, kidney cancer, hepatocellular carcinoma, thyroid cancer, gallbladder cancer, ovarian tumor, cervical cancer, gastric cancer, endometrial carcinoma, head and neck cancer, melanoma, neuroendocrine carcinoma, CNS cancer, brain tumor (e.g., glioma, degenerative rare glioblastoma, adult glioblastoma multiforme and adult spongioblastoma multiforme), bone carcinoma, soft tissue sarcoma, retinoblastoma, neuroblastoma, ascites, malignant pleural edema, mesothelioma, trophoblastic tumor, hemangiopericytoma, Kaposi's sarcoma, mucoid carcinoma, myxoid Carcinoma, round cell carcinoma, breast dermoid carcinoma, esophageal squamous cell carcinoma, oral carcinoma, adrenal cortical carcinoma, autoimmune diseases, atherosclerosis, arthritis (e.g., osteoarthritis or rheumatoid arthritis), inflammatory bowel disease (e.g., ulcerative colitis or Crohn's disease), peripheral vascular disease, cerebrovascular accident (stroke), chronic inflammation, Alzheimer's disease, neurodegenerative diseases or disorders, Aicardi-Goutieres syndrome, juvenile arthritis, osteoporosis, amyotrophic lateral sclerosis, multiple sclerosis, cardiac dysfunction, transplantation, or infectious diseases.

[0002] The invention further relates to pharmaceutical compositions comprising the compounds of the invention, to the use of said compounds in the manufacture of medicaments, and to methods of treating mammals, especially humans, by administering said compounds. [Background technology]

[0003] Humans and other mammals are exposed to bacteria and threats from pathogenic and non-pathogenic microorganisms, and have mechanisms of the immune system. The immune system relies on two major categories, namely, the innate (non-specific), general immune system and the adaptive (acquired), specialized immune system. Cells of the innate immune system sense infection through pattern recognition receptors (PRRs), including Toll-like receptors (TLRs), retinoic acid-inducible gene I-like receptors (RLRs), nucleotide oligomerization domain-like receptors (NLRs, also called NACHTs, LRRs, and PYD domain proteins), AIM2-like receptors (ALRs), and cytoplasmic DNA sensors (CDSs). PRRs sense ligands such as pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) released from damaged cells. PRR proteins are expressed by dendritic cells, monocytes, epithelial cells, and macrophages (Alberts B, et al, Innate immunity. Molecular biology of the cell. 4th ed New York: Gerland Science. (2002); Schroder K, et al, J. Cell. 140(6):821-32 (2010); Jounai N, et al, Front Cell Infect Microbiol. 2:168 (2013)). PRRs are activated by PAMP and DAMP ligands, which initiate signaling chain reactions leading to overexpression of pro-inflammatory cytokine and chemokine genes (Takeuchi O, et al, Cell. 140(6):805-20 (2010)).

[0004] The central signaling adaptor molecule stimulator of interferon genes (STING), also named TMEM 173, ERIS, MPYS, and MITA, is a ubiquitously expressed protein that is mainly confined to the cytoplasmic reticular (ER) membrane. STING can be recognized and activated by cyclic dinucleotides (CDNs) and aberrant DNA species or in the cytoplasm of cells, and plays a role in inducing pro-inflammatory cytokines, chemokines, and type I interferons (IFNs) in response (references [Wu JJ, et al, Med Res Rev. (2019); Ishikawa H, et al, Nature. 455 (7213): 674-678 (2008)]). STING is activated by cyclic GMP-AMP (cGAMP) synthase (cGAS), which generates 2',3'-cyclic GMP-AMP (2',3'-cGAMP) from cyclic-di-GMP (CDG) and cyclic-di-AMP (CDA). CDG and CDA are synthesized by bacteria such as Listeria monocytogenes. Upon binding to CDN, STING in turn recruits downstream Tank-binding kinase 1 (TBK1), which then phosphorylates STING and the transcription factor interferon regulatory factor 3 (IRF3), inducing type I IFNs and pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6), thus clearing pathogens and triggering long-lasting protective innate and adaptive immune system responses. Type I interferons play an important role in promoting cross-priming of CD8+ T cells and enhancing dendritic cells (references [Woodward JJ, et al, Science. 328(5986): 1703-1705 (2010); Chen J, et al, J Clin Invest. 129(10): 4224-4238 (2019)]).

[0005] Small changes in chemical synthesis and structure have substituted hydrophilic, unstable, and negatively charged CDNs for STING agonists. In addition, the phosphate moiety on CDNs has the disadvantage of being reduced by phosphoesterases and binding to STING. STING agonist compounds activate the innate immune system and antitumor immune responses through upregulation of IFN. Furthermore, STING activation within the tumor microenvironment (TME) drives the priming of T lymphocytes (reference [Yun LV, et al, Front Microbiol. 10 (2019)]).

[0006] This type of immune regulation is useful in autoinflammatory diseases, cancer, allergic diseases, neurodegenerative diseases, inflammatory diseases, amyotrophic lateral sclerosis, multiple sclerosis, and irritable bowel disease (references [Zitvogel L, et al, Nat Rev Immunol. 15(7):405-14(2015); Moisan J, et al, Am J Physiol. Lung Cell Mol. Physiol. 290(5):L987-95(2006); Cirulli ET, et al, Science. 347(6229):1436-41(2015)]).

[0007] On the other hand, increased type I IFN production is associated with chronic infections such as Mycobacteria, Franciscella, Chlamydia, and Plasmodium (Sebastian A, et al, J Immunol. 194(6):2455-2465(2015); Sebina I, et al, Immunology. 155(2):176-185(2018)]). In addition, it is also associated with patients with complex autoimmune diseases, including systemic lupus erythematosus (SLE), Aicardi-Goutières syndrome (AGS), and STING-associated infantile-onset vasculopathy (SAVI). Moreover, hyperactivity of the STING pathway induces cytokine storm and systemic inflammation. Thus, STING inhibitors could cure autoimmune diseases and treat patients with chronic activation of STING and pro-inflammatory cytokine production (Barber GN.et al,Nat Rev Immunol.15(12):760-70(2015);Gall A,et al,Immunity.36(1):120-31(2012);Crow YJ,et al,Nat Rev Immunol.15(7):429-40(2015);Rice GI,et al,J Clin Immunol.35(3):235-43(2015)).

[0008] STING agonist compounds treat cancer, superficial skin cancer, premalignant actinic keratosis, precancerous syndromes, infectious diseases, asthma and allergic rhinitis (references [Mathur V, et al, Neuron. 96(6):1290-1302(2017); Huber JP, et al, J Immunol. 185(2):813-7(2010)]). Thus, STING activity of agonists offers promising therapeutic effects in various diseases. Summary of the Invention [Means for solving the problem]

[0009] The present invention provides compounds of formula I, or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof:

[0010] [Chemical formula I] [ka]

[0011] During the ceremony,

[0012] Each R 1 are independently NH2, OH, and NHOR 4 or NHR 4 and;

[0013] Each R 2 are independently hydrogen, halogen, CF, -(C1-C6)alkyl, -O(C1-C6)alkyl, acyl, amino, substituted amino, cyano, acyloxy, or aryloxy;

[0014] R 3 is hydrogen, halogen, CF3, acyl, amino, substituted amino, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C3-C6)cycloalkyl, -(C1-C6)alkoxy, -(C1-C6)hydroxyalkyl, hetCyc 1 , hetCyc 2 , -(C1-C3) alkyl [hetCyc 1 ], -(C1-C3) alkyl [hetCyc 2 ], -(C1-C3) alkyl [hetAr 2 ], -(C1-C3) alkyl [hetAr 3 ], cyano, nitro, alkoxy, acyloxy or aryloxy;

[0015] R 4is H, trifluoromethyl, -(C1-C6)alkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C1-C3)alkyl[(C3-C6)cycloalkyl], -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl, -(C1-C6)alkylamine, -(C1-C6)hydroxyalkyl, -(C2-C6)dihydroxyalkyl, [(C1-C6)alkoxy](C1-C6)alkyl- or [(C1-C6)alkoxy]-[(C1-C6)alkoxy]-(C1-C6)alkyl-;

[0016] Each X 1 and X 2 are independently N or CR 5 and;

[0017] R 5 is hydrogen, halogen, OH, CF3, acyl, amino, substituted amino, -(C1-C6)alkyl, substituted (C1-C6)alkyl, -(C1-C6)haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C1-C6)alkyl, -O(C1-C6)hydroxyalkyl, -O(C2-C6)alkenyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[hetCyc 1 ], -O(C1-C6)alkyl[(C1-C6)alkoxy], -O(C2-C6)alkenyl[(C1-C6)alkoxy], -O(C2-C6)alkynyl[(C1-C6)alkoxy], -O(C1-C6)alkyl[(C3-C6)cycloalkyl], -O(C1-C3)alkyl[hetCyc 1 ], -O(C1-C3)alkyl[hetCyc 2 ] or -O(C2-C6)alkynyl[hetCyc 1 ] and;

[0018] hetCyc 1is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being selected from -(C1-C6) alkyl, -(C1-C6) alkoxy, OH, halogen, -C(O)R 6 , -CO2R 6 , -C(O)NR 6 R 7 , -S(O)NR 6 R 7 or -S(O)2R 6 optionally substituted with a substituent selected from

[0019] hetCyc 2 is a bridged eight-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom;

[0020] R 6 is H, -(C1-C6)alkyl, -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl, -(C1-C6alkyl)-NR 7 R 8 , -(C1-C6)hydroxyalkyl, -(C2-C6)dihydroxyalkyl, [(C1-C6)alkoxy](C1-C6)alkyl-, [(C1-C6)alkoxy]-[(C1-C6)alkoxy]-(C1-C6)alkyl-, hetCyc 1 , Ar 1 ,hetAr 2 or hetAr 3 and;

[0021] R 7 is H or -(C1-C6)alkyl;

[0022] NR 6 R 7 forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being selected from -(C1-C6) alkyl, OH, NH2, -(C1-C6) hydroxyalkyl, -(C1-C6) alkylamine, -COR 8 and -(C1-C3) alkylCO2R 8optionally substituted with one or more substituents independently selected from

[0023] R 8 is H, -(C1-C3)alkyl or -(C1-C3)hydroxyalkyl;

[0024] Ar 1 is phenyl optionally substituted with one or more substituents independently selected from (C1-C6)alkoxy, halogen, (C1-C6)alkyl, and CF3;

[0025] hetAr 2 is pyridyl optionally substituted with one or more substituents independently selected from halogen, CF, (C-C)alkyl, and (C-C)alkoxy;

[0026] hetAr 3 is a 5-membered heteroaryl having 2-3 ring heteroatoms independently selected from N, O and S, optionally substituted with (C1-C6)alkyl and OH;

[0027] Y 1 and Y 2 are independently N, O, S, and CR. 9 , N.R. 10 and;

[0028] R 9 is hydrogen, halogen, CF3, -(C1-C6)alkyl or -O(C1-C6)alkyl;

[0029] R 10 is hydrogen, -(C1-C6)alkyl or acyl;

[0030] A is -CH=CH-, -CC-, -CH2-, -CR 11 R 12 -, -C(O)NR 13 -, -C(O)NHOR 14 -, -NR 13 C(O)-, -NR 13 CO2-, -NR13 C(O)NR 13 -, -NR 13 -, -(C3-C7)cycloalkyl-, -O-, -S-, -S(O)- or -S(O)2-, optionally substituted -phenyl-, optionally substituted -(5- to 6-membered heteroaryl)- or optionally substituted -(5- to 6-membered heterocycloalkyl)-;

[0031] R 11 is selected from the group consisting of F, CF3, -(C1-C6)alkyl, substituted (C1-C6)alkyl, and cyano;

[0032] R 12 is H, F, CF3, -(C1-C6)alkyl;

[0033] R 11 and R 12 form together with the atom to which they are attached a 3- to 7-membered carbocyclic or heterocyclic ring;

[0034] R 13 is hydrogen, -(C1-C6)C alkyl, -(C1-C6)C fluoroalkyl, -(C1-C6) difluoroalkyl, or -(C1-C6) trifluoroalkyl;

[0035] R 14 is -(C1-C6)alkyl, -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl or [(C1-C6)alkoxy](C1-C6)alkyl-;

[0036] m is 0, 1, 2 or 3;

[0037] n is 0, 1, 2 or 3.

[0038] Compounds of formula I further include compounds of formulas II-a, II-b, and II-c, or pharma- ceutically acceptable salts thereof:

[0039] [Chemical formula II-a] [ka] [Chemical formula II-b] [ka] [Chemical formula II-c] [ka]

[0040] During the ceremony,

[0041] Each R 2 are independently hydrogen, halogen, CF, -(C1-C6)alkyl, -O(C1-C6)alkyl, acyl, amino, substituted amino, cyano, acyloxy, or aryloxy;

[0042] R 3 is hydrogen, halogen, CF3, acyl, amino, substituted amino, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C3-C6)cycloalkyl, -(C1-C6)alkoxy, -(C1-C6)hydroxyalkyl, hetCyc 1 , hetCyc 2 , -(C1-C3) alkyl [hetCyc 1 ], -(C1-C3) alkyl [hetCyc 2 ], -(C1-C3) alkyl [hetAr 2 ], -(C1-C3) alkyl [hetAr 3 ], cyano, nitro, alkoxy, acyloxy or aryloxy;

[0043] hetCyc 1 is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being selected from -(C1-C6) alkyl, -(C1-C6) alkoxy, OH, halogen, -C(O)R 6 , -CO2R 6 , -C(O)NR 6 R 7, -S(O)NR 6 R 7 or -S(O)2R 6 optionally substituted with a substituent selected from

[0044] hetCyc 2 is a bridged eight-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom;

[0045] Each X 1 and X 2 are independently N or CR 5 and;

[0046] R 5 is hydrogen, halogen, OH, CF3, acyl, amino, substituted amino, -(C1-C6)alkyl, substituted (C1-C6)alkyl, -(C1-C6)haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C1-C6)alkyl, -O(C1-C6)hydroxyalkyl, -O(C2-C6)alkenyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[hetCyc 1 ], -O(C1-C6)alkyl[(C1-C6)alkoxy], -O(C2-C6)alkenyl[(C1-C6)alkoxy], -O(C2-C6)alkynyl[(C1-C6)alkoxy], -O(C1-C6)alkyl[(C3-C6)cycloalkyl], -O(C1-C3)alkyl[hetCyc 1 ], -O(C1-C3)alkyl[hetCyc 2 ], or -O(C2-C6)alkynyl [hetCyc 1 ] and;

[0047] R 6 is H, -(C1-C6)alkyl, -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl, -(C1-C6alkyl)-NR 7 R 8, -(C1-C6)hydroxyalkyl, -(C2-C6)dihydroxyalkyl, [(C1-C6)alkoxy](C1-C6)alkyl-, [(C1-C6)alkoxy]-[(C1-C6)alkoxy]-(C1-C6)alkyl-, hetCyc 1 , Ar 1 ,hetAr 2 or hetAr 3 and;

[0048] R 7 is H or -(C1-C6)alkyl;

[0049] NR 6 R 7 forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being selected from -(C1-C6) alkyl, OH, NH2, -(C1-C6) hydroxyalkyl, -(C1-C6) alkylamine, -COR 8 and -(C1-C3) alkylCO2R 8 optionally substituted with one or more substituents independently selected from

[0050] R 8 is H, -(C1-C3)alkyl or -(C1-C3)hydroxyalkyl;

[0051] Ar 1 is phenyl optionally substituted with one or more substituents independently selected from (C1-C6)alkoxy, halogen, (C1-C6)alkyl, and CF3;

[0052] hetAr 2 is pyridyl optionally substituted with one or more substituents independently selected from halogen, CF, (C-C)alkyl, and (C-C)alkoxy;

[0053] hetAr 3 is a 5-membered heteroaryl having 2-3 ring heteroatoms independently selected from N, O and S, optionally substituted with (C1-C6)alkyl and OH;

[0054] Y 1 and Y 2 are independently N, O, S, and CR. 9 , N.R. 10 and;

[0055] R 9 is hydrogen, halogen, CF3, -(C1-C6)alkyl or -O(C1-C6)alkyl;

[0056] R 10 is hydrogen, -(C1-C6)alkyl or acyl.

[0057] Compounds of formula I further include compounds of formulas III-a, III-b, III-c, and III-d, or pharma- ceutically acceptable salts thereof:

[0058] [Chemical formula III-a] [ka] [Chemical formula III-b] [ka] [Chemical formula III-c] [ka] [Chemical formula III-d] [ka]

[0059] During the ceremony,

[0060] Z is -R 6 , -OR 6 , -NR 6 R 7 , -(C1-C6)alkyl, -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl, -(C1-C6alkyl)-NR 7 R 8, -(C1-C6)hydroxyalkyl, -(C2-C6)dihydroxyalkyl, [(C1-C6)alkoxy](C1-C6)alkyl- or [(C1-C6)alkoxy]-[(C1-C6)alkoxy]-(C1-C6)alkyl-;

[0061] L is CH or N;

[0062] Each X 1 and X 2 are independently N or CR 5 and;

[0063] R 5 is hydrogen, halogen, OH, CF3, acyl, amino, substituted amino, -(C1-C6)alkyl, substituted (C1-C6)alkyl, -(C1-C6)haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C1-C6)alkyl, -O(C1-C6)hydroxyalkyl, -O(C2-C6)alkenyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[(C4-C6)hydroxy], -O(C2-C6)alkynyl[hetCyc 1 ], -O(C1-C6)alkyl[(C1-C6)alkoxy], -O(C2-C6)alkenyl[(C1-C6)alkoxy], -O(C2-C6)alkynyl[(C1-C6)alkoxy], -O(C1-C6)alkyl[(C3-C6)cycloalkyl], -O(C1-C3)alkyl[hetCyc 1 ], -O(C1-C3)alkyl[hetCyc 2 ], or -O(C2-C6)alkynyl [hetCyc 1 ] and;

[0064] R 6 is H, -(C1-C6)alkyl, -(C1-C6)fluoroalkyl, -(C1-C6)difluoroalkyl, -(C1-C6)trifluoroalkyl, -(C1-C6alkyl)-NR 7 R 8, -(C1-C6)hydroxyalkyl, -(C2-C6)dihydroxyalkyl, [(C1-C6)alkoxy](C1-C6)alkyl-, [(C1-C6)alkoxy]-[(C1-C6)alkoxy]-(C1-C6)alkyl-, hetCyc 1 , Ar 1 ,hetAr 2 or hetAr 3 and;

[0065] R 7 is H or -(C1-C6)alkyl;

[0066] NR 6 R 7 forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being selected from -(C1-C6) alkyl, OH, NH2, -(C1-C6) hydroxyalkyl, -(C1-C6) alkylamine, -COR 8 and -(C1-C3) alkylCO2R 8 optionally substituted with one or more substituents independently selected from

[0067] R 8 is H, -(C1-C3)alkyl or -(C1-C3)hydroxyalkyl;

[0068] hetCyc 1 is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being selected from -(C1-C6) alkyl, -(C1-C6) alkoxy, OH, halogen, -C(O)R 6 , -CO2R 6 , -C(O)NR 6 R 7 , -S(O)NR 6 R 7 or -S(O)2R 6 optionally substituted with a substituent selected from

[0069] hetCyc 2 is a bridged eight-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom;

[0070] Ar 1 is phenyl, optionally substituted with one or more substituents independently selected from (C1-C6)alkoxy, halogen, (C1-C6)alkyl, and CF3;

[0071] hetAr 2 is pyridyl optionally substituted with one or more substituents independently selected from halogen, CF, (C-C)alkyl, and (C-C)alkoxy;

[0072] hetAr 3 is a 5-membered heteroaryl having 2-3 ring heteroatoms independently selected from N, O and S, and optionally substituted with (C1-C6)alkyl and OH.

[0073] The compounds of the present invention are agents of stimulator of interferon genes (STING) transformation and therefore are useful in treating breast cancer, ovarian cancer, prostate cancer, testicular cancer, urothelial carcinoma, bladder cancer, non-small cell lung cancer, small cell lung cancer, sarcoma, colorectal adenocarcinoma, gastrointestinal stromal tumor, gastroesophageal carcinoma, colon cancer, pancreatic cancer, renal cancer, hepatocellular carcinoma, malignant mesothelioma, leukemia, lymphoma, myelodysplastic syndrome, multiple myeloma, transitional cell carcinoma, neuroblastoma, plasma cell neoplasm, Wilms' tumor, hepatocellular carcinoma, urinary tract cancer, bladder carcinoma, colorectal cancer, colon cancer, breast cancer, prostate cancer, renal cancer, hepatocellular carcinoma, thyroid cancer, gallbladder cancer, ovarian tumor, cervical cancer, gastric cancer, endometrial carcinoma, head and neck cancer, melanoma, neuroendocrine carcinoma, CNS cancer, brain tumors (e.g., glioma, degenerative rare glioblastoma, adult and adult spongioblastoma multiforme), bone carcinoma, soft tissue sarcoma, retinoblastoma, neuroblastoma, ascites, malignant pleural edema, mesothelioma, trophoblastic tumor, hemangiopericytoma, Kaposi's sarcoma, mucinous carcinoma, round cell carcinoma, breast dermoid carcinoma, esophageal squamous cell carcinoma, oral carcinoma, adrenal cortical carcinoma, autoimmune disease, atherosclerosis, arthritis (e.g., osteoarthritis or rheumatoid arthritis), inflammatory bowel disease (e.g., ulcerative colitis or Crohn's disease), peripheral vascular disease, cerebrovascular accident (stroke), chronic inflammation, Alzheimer's disease, neurodegenerative disease or disorder, Aicardi-Goutieres syndrome, juvenile arthritis, osteoporosis, amyotrophic lateral sclerosis, multiple sclerosis, cardiac dysfunction, transplantation, or infection.

[0074] In another aspect, the present invention relates to a pharmaceutical composition comprising an effective amount of a compound of formula I, or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof. In some embodiments, the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier, adjuvant, and / or excipient. In some embodiments, such a composition may include any of preservatives, preparations for delayed absorption, fillers, binders, adsorbents, buffers, disintegrants, solubilizers, and other carriers, adjuvants, and / or excipients as inactive ingredients. The composition may be formulated in a manner well known in the art.

[0075] In some embodiments, the present invention relates to a method of treating a disease in an individual comprising administering to said individual a therapeutically effective amount of a composition comprising a compound of Formula I or a pharma-ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof.

[0076] In another aspect, the present invention relates to a method of treating a disorder in a mammal comprising administering to said mammal a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof.

[0077] In another aspect, the present invention relates to a method of treating a disorder in a human comprising administering to said human a therapeutically effective amount of a compound of formula I or a pharma-ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof.

[0078] In another aspect, the present invention provides a method for treating breast cancer, ovarian cancer, prostate cancer, testicular cancer, urothelial carcinoma, bladder cancer, non-small cell lung cancer, small cell lung cancer, sarcoma, colorectal adenocarcinoma, gastrointestinal stromal tumor, ovarian cancer, prostate cancer, testicular cancer, urothelial carcinoma, bladder cancer, sarcoma, colorectal adenocarcinoma, gastrointestinal stromal tumor, ovarian cancer, prostate cancer, urothelial carcinoma, bladder cancer, urothelial carcinoma ... Gastroesophageal carcinoma, colorectal cancer, pancreatic cancer, renal cancer, hepatocellular carcinoma, malignant mesothelioma, leukemia, lymphoma, myelodysplastic syndrome, multiple myeloma, transitional cell carcinoma, neuroblastoma, plasma cell neoplasm, Wilms' tumor, hepatocellular carcinoma, urinary tract cancer, bladder carcinoma, colorectal cancer, colon cancer, breast cancer, prostate cancer, renal cancer, hepatocellular carcinoma, thyroid cancer, gallbladder cancer, ovarian tumor, cervical cancer, gastric cancer, endometrial carcinoma, head and neck cancer, melanoma, neuroendocrine carcinoma, C NS cancer, brain tumors (e.g., glioma, degenerative rare glioblastoma, adult glioblastoma multiforme and adult spongioblastoma multiforme), bone carcinoma, soft tissue sarcoma, retinoblastoma, neuroblastoma, ascites, malignant pleural edema, mesothelioma, trophoblastic tumor, hemangiopericytoma, Kaposi's sarcoma, mucinous carcinoma, round cell carcinoma, breast dermoid carcinoma, esophageal squamous cell carcinoma, oral carcinoma, adrenal cortical carcinoma, autoimmune diseases, atherosclerosis, The present invention relates to methods of treating arthritis (e.g., osteoarthritis or rheumatoid arthritis), inflammatory bowel disease (e.g., ulcerative colitis or Crohn's disease), peripheral vascular disease, cerebrovascular accidents (stroke), chronic inflammation, Alzheimer's disease, a neurodegenerative disease or disorder, Aicardi-Goutières syndrome, juvenile arthritis, osteoporosis, amyotrophic lateral sclerosis, multiple sclerosis, cardiac dysfunction, transplantation or infectious disease, a condition or disorder.

[0079] In another aspect, the present invention relates to a method of treating a disorder or condition modulated by Stimulator of Interferon Genes (STING) transformation in a mammal, including a human, comprising administering to said mammal an amount of a compound of formula I, or a pharma- ceutically acceptable salt, ester, prodrug, solvate, such as a hydrate, polymorph, or tautomer thereof, effective to modulate the chain reaction.Appropriate dosages for a particular patient can be determined according to methods known to those skilled in the art.

[0080] In another aspect, the present invention relates to the use of the compound of formula I, or its pharma- ceutically acceptable salt, ester, prodrug, solvate, e.g., hydrate, polymorph, or tautomer, in the manufacture of a pharmaceutical composition.The pharmaceutical composition can be used to treat disorders or conditions regulated by stimulator of interferon genes (STING) transformation in mammals, including humans.The pharmaceutical composition is useful for treating cancer and other inflammation.

[0081] In another aspect, the invention relates to pharmaceutical compositions comprising a compound of formula I, or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof. In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. In further or additional embodiments, the pharmaceutical composition is in the form of tablets, capsules, pills, powders, sustained release formulations, solutions, and suspensions. In some embodiments, the pharmaceutical composition is in a form suitable for parenteral injection, such as a sterile solution, suspension, or emulsion, a form suitable for topical administration as an ointment or cream, or a form suitable for rectal administration as a suppository. In further or additional embodiments, the pharmaceutical composition is in a unit dosage form suitable for single administration of a precise dosage. In further or additional embodiments, the amount of the compound of formula I ranges from about 0.001 to about 1000 mg / kg body weight / day. In further or additional embodiments, the amount of the compound of formula I ranges from about 0.5 to about 50 mg / kg body weight / day.

[0082] In another aspect, the present invention relates to a process for preparing a compound of Formula I or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer, or prodrug thereof.

[0083] technical challenges The present invention provides novel substituted heterocyclic compounds represented by formula I, or pharma-ceutically acceptable salts, solvates, polymorphs, esters, tautomers or prodrugs thereof, and compositions comprising the compounds.

[0084] The present invention also provides pharmaceutical compositions comprising the compounds of the present invention.

[0085] The invention further provides the use of said compounds in the manufacture of a medicament, and a method of treating a mammal, particularly a human, by administering said compounds.

[0086] Means for solving the problem The novel features of the invention are set forth with particularity in the appended claims. The features and advantages of the present invention will be more readily understood by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized.

[0087] While preferred embodiments of the present invention have been shown and described herein, such embodiments are provided by way of example only. It should be understood that various alternatives to the embodiments of the present invention described herein may be used in practicing the present invention. Those skilled in the art will recognize that various changes, modifications, and substitutions are possible without departing from the invention. The following claims are intended to define the scope of the aspects of the present invention, and it is intended to cover methods and structures within the scope of such claims and their equivalents.

[0088] The section headings used herein are for organizational purposes only and should not be construed as limiting the claimed subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and papers, are expressly incorporated herein by reference in their entirety for any purpose. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0089] Specific Chemical Terms

[0090] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which the claimed subject matter belongs. Unless otherwise stated, all patents, patent applications, and published materials mentioned throughout the disclosure of this specification are incorporated by reference in their entirety. In the event that there are multiple definitions for terms herein, the definitions in this section shall prevail. When a URL or other such identifier or address is referenced, it is understood that such identifiers may vary and specific information on the Internet may vary, but equivalent information may be found by searching the Internet or other appropriate reference. Reference thereto evidences the availability and public dissemination of such information.

[0091] The foregoing general description and the following detailed description are merely exemplary and explanatory and should not be construed as limiting any claims that may be made. In this application, singular terms include plurals unless otherwise specified. As used in the specification and the appended claims, it should be noted that the singular includes plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or" means "and / or" unless otherwise specified. Furthermore, the use of different forms of terms such as "starting with" and "starting with" is not intended to be limiting. Similarly, the use of different forms of terms such as "including" and "including" is not intended to be limiting.

[0092] Definitions of standard chemical terms are given in the literature [Carey and Sundberg, "ADVANCED ORGANIC CHEMISTRY 4 TH"Ed." Vols. A (2000) and B (2001), Plenum Press, New York]. Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, IR and UV / Vis spectroscopy, and pharmacology, which are within the skill of the art, are used. Unless otherwise defined, the nomenclature used in connection with analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein, and the laboratory procedures and techniques are known in the art. Standard techniques for chemical synthesis, chemical analysis, pharmaceutical manufacture, dosage forms, and patient delivery and treatment may be used. Reactions and purification techniques may be performed, for example, using kits according to manufacturer's specifications, as commonly practiced in the art, or as described herein. In general, the techniques and procedures described above may be performed according to conventional methods well known to those of skill in the art and as described in the various general and more specific references cited and discussed throughout this specification. Throughout this specification, groups and substituents thereof may be selected by one of skill in the art to provide stable moieties and compounds.

[0093] Where substituents are specified by their conventional chemical formula written from left to right, they equally include the chemically identical substituents that result from writing the structure from right to left. As a non-limiting example, CH2O is the same as OCH2.

[0094] Unless otherwise noted, general chemical terms such as, but not limited to, "alkyl," "amine," and "aryl" are used interchangeably with their optionally substituted forms. For example, "alkyl" as used herein includes optionally substituted alkyl.

[0095] The compounds presented herein may have one or more stereocenters, each of which may be present in the R or S configuration, or a combination thereof. Similarly, the compounds presented herein may have one or more double bonds, each of which may be present in the E (trans) or Z (cis) configuration, or a combination thereof. The presentation of a particular stereoisomer, positional isomer, partial stereoisomer, enantiomer, or epimer should be understood to include all possible stereoisomers, positional isomers, partial stereoisomers, enantiomers, or epimers, and mixtures thereof. Thus, the compounds presented herein include all individual sequence stereoisomers, positional isomers, partial stereoisomers, enantiomers, and epimeric forms, as well as the corresponding mixtures thereof. Techniques for inverting or leaving unchanged specific stereocenters, and for separating mixtures of stereoisomers, are well known to those skilled in the art, and it is well within the ability of the skilled artisan to select the method appropriate for a particular situation. See, for example, the literature [Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; and Heller, Acc. Chem. Res. 1990, 23, 128].

[0096] The term "bond" or "single bond" refers to a chemical bond between two atoms or two moieties when the atoms connected by the bond are considered to be part of a larger structure.

[0097] The term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes cases where said event or circumstance occurs and does not occur. For example, "optionally substituted alkyl" means "alkyl" or "substituted alkyl" as defined below. Furthermore, an optionally substituted group may be unsubstituted (e.g., CH2CH3), fully substituted (e.g., CF2CF3), monosubstituted (e.g., CH2CH2F), or substituted at any level between fully and monosubstituted (e.g., CH2CHF2, CF2CH3, CFHCHF2, etc.). For any group containing one or more substituents, one of skill in the art will understand that such group is not intended to introduce any substitution or substitution pattern that is sterically impractical and / or synthetically impractical (e.g., substituted alkyl includes optionally substituted cycloalkyl groups, which are ultimately defined to include optionally substituted alkyl groups, potentially limitless). Thus, any described substituent should be understood to generally have a maximum molecular weight of about 1,000 daltons, more typically about 500 daltons or less (except where a macromolecular substituent is expressly intended, e.g., polypeptides, polysaccharides, polyethylene glycol, DNA, RNA, etc.).

[0098] As used herein, C1-Cn includes C1-C2, C1-C3, ...C1-Cn. By way of example, a group designated as "C1-C4" indicates that there are 1 to 4 carbon atoms in the moiety, i.e., groups containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms, as well as the ranges C1-C2 and C1-C3. Thus, by way of example, "C1-C4 alkyl" indicates that there are 1 to 4 carbon atoms in the alkyl group, i.e., the alkyl group is selected from among methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and t-butyl. Numeric ranges such as "1 to 10" refer to the respective integers within the given range given whenever described herein, e.g., "1 to 10 carbon atoms" means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0099] As used herein, the term "heteroatom" or "hetero", alone or in combination, refers to an atom other than carbon and hydrogen. Heteroatoms are independently selected from among oxygen, nitrogen, sulfur, phosphorus, silicon, selenium and tin, but are not limited to these atoms. In embodiments where two or more heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other.

[0100] The term "alkyl," as used herein, alone or in combination, refers to an optionally substituted straight-chain or optionally substituted branched-chain saturated hydrocarbon monoradical having 1 to about 10 carbon atoms, more preferably 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-amyl and hexyl, as well as longer alkyl groups such as heptyl, octyl, and the like. Each time a numerical range such as "C1-C6 alkyl" or "C1_6 alkyl" appears herein, it means that the alkyl group can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms, but this definition also encompasses the occurrence of the term "alkyl" where no numerical range is given.

[0101] The term "aliphatic," as used herein, alone or in combination, refers to an optionally substituted straight or branched chain, acyclic, saturated, partially unsaturated or fully unsaturated non-aromatic hydrocarbon. Thus, the term generically includes alkyl, alkenyl and alkynyl groups.

[0102] The terms "cycle", "cyclic", "ring", "membered" and "-membered ring" as used herein, alone or in combination, refer to any covalently enclosed structure, including alicyclic, heterocyclic, aromatic, heteroaromatic, and polycyclic fused or non-fused ring systems as described herein. The ring may be optionally substituted. The ring may form part of a fused ring system. The term "-membered" is meant to indicate the number of skeletal atoms that constitute the ring. Thus, by way of example, cyclohexane, pyridine, pyran and pyrimidine are six-membered rings, and cyclopentane, pyrrole, tetrahydrofuran and thiophene are five-membered rings.

[0103] The term "cycloalkyl," as used herein, alone or in combination, refers to an optionally substituted saturated, hydrocarbon monoradical ring containing from 3 to about 15 annular carbon atoms or from 3 to about 10 annular carbon atoms, but which may contain additional non-annular carbon atoms as substituents (e.g., methylcyclopropyl).

[0104] Non-limiting examples of "cycloalkyl" include aziridinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl, quinolizinyl, and the like. The term also includes all rings of carbohydrates, including but not limited to monosaccharides, disaccharides and oligosaccharides.

[0105] The term "aromatic" as used herein refers to a non-uniform planar, cyclic or polycyclic ring moiety in an electron system containing 4n+2n electrons, where n is an integer. Aromatic rings can be formed by 5, 6, 7, 8, 9, or more than 9 atoms. Aromatics may be optionally substituted and may be monocyclic or fused-ring polycyclic. The term "aromatic" includes both all-carbon rings (e.g., phenyl) and rings containing one or more heteroatoms (e.g., pyridine).

[0106] Specific pharmaceutical terms

[0107] The terms "subject," "patient," or "individual," as used herein with respect to an individual having a disorder, condition, or the like, include mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates, such as chimpanzees, and other ape and monkey species; farm animals, such as cows, horses, sheep, goats, pigs; livestock, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, and the like. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0108] The term "treat" or "treatment" as used herein and other grammatical equivalents include alleviating, attenuating or ameliorating the symptoms of a disease or condition, preventing additional symptoms, improving or preventing the underlying metabolic cause of a symptom, inhibiting a disease or condition, e.g., preventing the onset of a disease or condition, relieving a disease or condition, inducing inhibition of a disease or condition, alleviating a condition caused by a disease or condition, or interrupting the symptoms of a disease or condition, and is intended to include prevention. The term further includes achieving a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit refers to the eradication or amelioration of the underlying disease being treated. A therapeutic benefit is also achieved when a patient may still suffer from the underlying disease, but one or more of the physiological symptoms associated with the underlying disease are eradicated or ameliorated, and an improvement is observed in the patient. For a prophylactic benefit, the composition can be administered to a patient at risk of developing a particular disease, even if the disease has not been diagnosed, or to a patient who has reported one or more of the physiological symptoms of the disease.

[0109] The terms "effective amount", "therapeutically effective amount" or "pharmaceutical effective amount" as used herein refer to a sufficient amount of at least one formulation or compound administered such that one or more of the symptoms of the disease or condition being treated are alleviated to some extent. The result may be a reduction and / or alleviation of the signs, symptoms or causes of the disease, or any other favorable alteration of a biological system. For example, a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof is an amount of a formulation of the present invention that, when administered to a human in need thereof, is sufficient to modulate the activity of STING such that a disease state mediated by STING activity is reduced, alleviated, or prevented. The appropriate "effective" amount in any individual can be determined using techniques such as dose escalation studies.

[0110] The terms "administer", "administration" and the like used herein refer to methods that can be used to deliver a compound or composition to a desired biological site of action. Such methods include, but are not limited to, oral route, intraduodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, or infusion), topical and rectal administration. Those skilled in the art are familiar with administration techniques that can be used with the compounds and methods described herein, for example, as discussed in the literature [Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.]. In a preferred embodiment, the compounds and compositions described herein are administered orally.

[0111] The term "acceptable" as used herein with respect to a dosage form, composition, or ingredient means that it does not have a continuing detrimental effect on the general health of the subject being treated.

[0112] The term "pharmaceutical acceptable" as used herein refers to a substance, such as a carrier or diluent, that does not interfere with the biological activity or properties of the compounds described herein and is relatively non-toxic, i.e., the substance can be administered to an individual without causing undesired biological effects or interacting in a deleterious manner with any components included in the composition.

[0113] The term "pharmaceutical composition" as used herein refers to a biologically active compound optionally mixed with at least one pharma- ceutically acceptable chemical component such as, but not limited to, a carrier, stabilizer, diluent, dispersant, suspending agent, thickener and / or excipient.

[0114] The term "carrier," as used herein, refers to relatively non-toxic chemical compounds or formulations that facilitate the incorporation of a compound into cells or tissues.

[0115] The term "agonist" as used herein refers to a molecule, such as a compound, a drug, an enzyme activator, or a hormone modulating agent, that enhances the activity of another molecule or the activity of a receptor site.

[0116] The term "modulate" as used herein means to interact with a target directly or indirectly to alter the activity of the target, including, by way of example, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.

[0117] The term "modulator" as used herein refers to a molecule that interacts with a target either directly or indirectly, including, but not limited to, the interactions of an agonist and an antagonist.

[0118] The term "pharmaceutical acceptable salts" as used herein refers to salts that maintain the biological effectiveness of the free acids and bases of a particular compound and are not biologically or otherwise undesirable. The compounds described herein may have acidic or basic groups and therefore can react with any of a number of inorganic or organic salts, and inorganic and organic acids to form pharmaceutical acceptable salts. Such salts can be prepared in the same system during the final isolation and purification of the compounds of the present invention, or can be prepared by separately reacting the compound purified in the form of a free base with an appropriate organic or inorganic acid and isolating the salt formed thereby.Examples of pharma- ceutically acceptable salts include salts prepared by reaction of the compounds described herein with an inorganic or organic acid or base, such as acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butyne-1,4-dioate, camphorate, camphosulfonate, caprylate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, decanoate, digluconate, dihydrogen phosphate, dinitrobenzoate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexyne-1,6-dioate, hydroxybenzoate, hydroxybutyrate, hydrochloride ... Examples of suitable sulphates include chloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maliate, malonate, methanesulfonate, mandelate, metaphosphate, methoxybenzoate, methylbenzoate, monohydrogen phosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, suberate, sebacate, sulfonate, tartrate, thiocyanate, tosylate, undeconate, and xylenesulfonate. Other acids, such as oxalic acid, although not themselves pharma- ceutically acceptable, can be used to prepare salts useful as intermediates for obtaining the compounds of the invention and their pharma- ceutically acceptable acid addition salts (see, for example, Berge et al., J. Pharm. Sci. 1977, 66, 1-19).Additionally, the compounds described herein, which may contain free acid groups, can be reacted with a suitable base, such as hydroxides, carbonates, or bicarbonates of pharmaceutically acceptable metal cations, with ammonia, or with pharmaceutically acceptable organic primary, secondary, or tertiary amines. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Illustrative examples of bases include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, and the like. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like. The compounds described herein should also be understood to include the quaternization of any basic nitrogen-containing groups that may be included. Water- or oil-soluble or dispersible products can be obtained by such quaternization. See, for example, Berge et al., supra.

[0119] The term "solvate" as used herein refers to a combination of a compound of the present invention and a solvent molecule formed by solvation. In some circumstances, the solvate refers to a hydrate, i.e., the solvent molecule is a water molecule, and the combination of a compound of the present invention and water forms a hydrate.

[0120] The terms "polymorph" or "polymorphic" as used herein refer to compounds of the present invention that exist in different crystal lattice forms.

[0121] The term "ester" as used herein refers to a derivative of a compound of the present invention derived from an oxoacid group and a hydroxyl group, either of which can be present in a compound of the present invention.

[0122] The term "tautomers" as used herein refers to isomers that are readily interconverted from the compounds of the invention, for example, by migration of a hydrogen atom or proton.

[0123] The term "pharmaceutical acceptable derivative or prodrug" as used herein refers to any pharmaceutical acceptable salt, ester, salt of an ester or other derivative of a compound of the invention, which upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the invention, or a pharmaceutical active metabolite or residue thereof. Particularly advantageous derivatives or prodrugs are those which increase the bioavailability of a compound of the invention when such compound is administered to a patient (e.g., by allowing an orally administered compound to be absorbed more readily into the blood) or which enhance the delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system).

[0124] Pharmaceutically acceptable prodrugs of the compounds described herein include, but are not limited to, esters, carbonates, thiocarbonates, N-acyl derivatives, N-acyloxyalkyl derivatives, quaternary derivatives of tertiary amines, N-Mannich bases, Schiff bases, amino acid conjugates, phosphate esters, metal salts and sulfonate esters. Various forms of prodrugs are widely known in the art. See, for example, Design of Prodrugs, Bundgaard, A. Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.; Academic, 1985, vol. 42, p. 309-396; Bundgaard, H. "Design and Application of Prodrugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and H. Bundgaard, Ed, 1991, Chapter 5, p. 113-191; and Bundgaard, H., Advanced Drug Delivery Review, 1992, 8, 1-38, each of which is incorporated herein by reference. Prodrugs described herein include, but are not limited to, the following groups and combinations of these groups: amine derived prodrugs. Hydroxy prodrugs include, but are not limited to, acyloxyalkyl esters, alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters and disulfide-containing esters.

[0125] The terms "enhance" or "enhancing" as used herein means to increase or prolong either the potency or duration of a desired effect. Thus, in enhancing the effect of a therapeutic agent, the term "enhancing" refers to the ability to increase or prolong, in potency or duration, the effect of other therapeutic agents on a system.

[0126] An "enhancing-effective amount," as used herein, refers to an amount adequate to enhance the effect of another therapeutic agent in a desired system.

[0127] The terms "pharmaceutical combination", "additional therapeutic administration", "additional therapeutic agent administration" and the like as used herein refer to pharmaceutical therapy resulting from mixing or combining more than one active ingredient, including both fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that at least one of the compounds described herein and at least one co-formulation are administered simultaneously to a patient in the form of a single entity or single dosage. The term "non-fixed combination" means that at least one of the compounds described herein and at least one co-formulation are administered simultaneously, together, or sequentially as separate entities with various intervening time limits to a patient, such administration providing effective levels of two or more compounds in the patient's body. This also applies to cocktail therapy, for example, administration of three or more active ingredients.

[0128] As used herein, the terms "co-administered," "administered together with," and their grammatical equivalents, etc., are meant to include administration of a selected therapeutic agent to a single patient, and are intended to include therapeutic regimes in which the therapeutic agents are administered by the same or different routes of administration, or at the same or different times. In some embodiments, the compounds described herein are administered simultaneously with other formulations. These terms include administering two or more formulations to an animal, where both the formulations and / or their metabolites are present in the animal at the same time. These include administration of separate compositions simultaneously, administration of separate compositions at different times, and / or administration of a composition in which both formulations are present. Thus, in some embodiments, the compounds of the present invention and other formulations are administered as a single composition.

[0129] The term "metabolite" as used herein refers to a derivative of a compound that is formed when the compound is metabolized.

[0130] The term "active metabolite" as used herein refers to a biologically active derivative of a compound that is formed when the compound is metabolized.

[0131] The term "metabolized" as used herein refers to the sum of the processes by which a particular substance is transformed by an organism, including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes. Thus, enzymes can cause specific structural modifications to compounds. For example, cytochrome P450 catalyzes various oxidation and reduction reactions, while uridine diphosphate glucuronyltransferase catalyzes the transfer of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups. Additional information on metabolism can be obtained from the literature [The Pharmacological Basis of Therapeutics, 9th Edition, McGraw-Hill (1996)].

[0132] Description of the embodiments Experimental Part

[0133] NMR spectra were recorded in DMSO-d6, MeOH-d4, and CDCl3 solutions in 5-mm od tubes (Norell, Inc. 507-HP) at 30 °C. 1 Analytes were collected on a JEOL at 400 MHz for H. Chemical shifts (δ) are relative to tetramethylsilane (TMS = 0.00 ppm) and are expressed in ppm. LC / MS was measured on an ISQ EM ion trap mass spectrometer, Thermo Fisher Vanquish Flex (column: hypersil Gold (C18, φ2.1x50mm, 1.9μm, 120Å, 30°C) operated in ESI(+) ionization mode; flow rate = 0.5mL / min. Mobile phase = 0.01% heptafluorobutyric acid (HFBA) and 1.0% isopropyl alcohol (IPA) in water or CH3CN.

[0134] [ka]

[0135] Intermediate 1: Methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride

[0136]

[0137] Step A: Methyl (E)-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate

[0138] To a solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate (5.00 g, 20.4 mmol) in n-BuOH (50 mL) at room temperature, tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate hydrochloride (4.53 g, 20.4 mmol) and DIPEA (17.7 mL, 102 mmol) were added. The reaction mixture was heated at 120° C. for 12 h. After concentration in vacuo, the residue was dissolved in EtOAc and washed with saturated aqueous NaHCO3 solution. The separated aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by recrystallization from MeOH to provide the title compound (5.56 g, 66%) as a yellow solid. 1 HNMR(400MHz,DMSO-d6):δ8.17(1H,d,J=1.9Hz),8.02(1H,t,J=6.2Hz),7.43(1H,d,J=1.9Hz),6.94(1H,t,J=5.9H z),5.52(2H,t,J=3.0Hz),4.12(2H,dd,J=6.1,3.0Hz),3.89(3H,s),3.83(3H,s),3.50-3.43(2H,m),1.35(9H,s). LC-MS:m / z=396.10[M+H] + .

[0139] Step B: Methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride

[0140] To a solution of methyl (E)-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (1.00 g, 2.53 mmol) in MeOH (6.8 mL) at room temperature was added HCl (4 M in dioxane, 6.3 mL, 25 mmol). The reaction mixture was stirred at room temperature overnight. After concentration in vacuo, the residual solid was suspended in Et2O, collected by filtration, washed with Et2O and then dried under vacuum to provide the title compound (827 mg, 99%) as an orange solid. 1H NMR(400MHz,DMSO-d6):δ8.19(1H,d,J=1.6Hz),8.14(1H,t,J=7.2Hz),7.81(3H,brs),7.46(1H,d,J=2.0H z),5.87-5.82(1H,m),5.63-5.58(1H,m),4.21(2H,t,J=6.0Hz),3.90(3H,s),3.84(3H,s),3.40(2H,brs).

[0141] [ka]

[0142] Intermediate 2: (E)-2-(4-aminobut-2-en-1-yl)isoindoline-1,3-dione

[0143] [ka]

[0144] Step A: (E)-2-(4-bromobut-2-en-1-yl)isoindoline-1,3-dione

[0145] To a solution of potassium phthalimide (10.0 g, 54.0 mmol) in DMF (56 mL) at room temperature was added (E)-1,4-dibromobut-2-ene (34.6 g, 162 mmol). The reaction mixture was stirred at room temperature for 22 h. After addition of cold water, the precipitated solid was collected by filtration. The solid was purified by column chromatography on SiO2 (hexane:EtOAc = 4:1 to 1:1) to provide the title compound (9.86 g, 65%) as a white solid. 1 H NMR (400MHz, CDCl3): δ7.88-7.83(2H,m),7.75-7.70(2H,m),5.98-5.80(1H,m),5.86-5.79(1H,m),4.30(2H,dd,J=0.8,6.0Hz),3.90(2H,d,J=7.6Hz).

[0146] Step B: (1s,3R,5S)-1-((E)-4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)-1,3,5,7-tetraaza-adamantan-1-ium bromide

[0147] To a solution of (E)-2-(4-bromobut-2-en-1-yl)isoindoline-1,3-dione (9.86 g, 35.2 mmol) in CHCl3 (98 mL) at room temperature was added hexamethylenetetraamine (7.40 g, 52.8 mmol). The reaction mixture was stirred at room temperature for 26 h. The precipitated solid was collected by filtration, washed with CHCl3, and dried under vacuum to provide the title compound (14.4 g, 97%) as a white powder. 1 H NMR(400MHz,CD3OD):δ7.90-7.82(4H,m),6.19-6.12(1H,m),5.93-5.85(1H,m),5.06(6H ,s),4.73-4.70(3H,m),4.55-4.52(3H,m),4.41(2H,d,J=5.6Hz),3.47(2H,d,J=7.6Hz).

[0148] Step C: (E)-2-(4-aminobut-2-en-1-yl)isoindoline-1,3-dione

[0149] To a solution of (1s,3R,5S)-1-((E)-4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)-1,3,5,7-tetraaza-adamantan-1-ium bromide (9.80 g, 23.3 mmol) in EtOH (192 mL) at 0° C. was added concentrated HCl (9.80 mL, 118 mmol). The reaction mixture was refluxed for 2 h. After concentration in vacuum, the residue was purified by crystallization from Et2O / MeOH to provide the HCl salt form of the title compound (5.89 g, quant.) as a white solid. The salt compound (5.89 g, 23.2 mmol) was dissolved in DCM / MeOH and then basified with 1N aqueous NaOH and saturated aqueous NaHCO3 until pH 8. The separated organic layer was washed with brine, dried over Na2SO4, filtered and concentrated in vacuum. The residue was concentrated in vacuo to provide the title compound (3.17 g, 63%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ7.91-7.85(4H,m),5.91-5.59(2H,m),4.20(1H,dd,J=5.6,1.2Hz),3.43-3.38(2H,m).

[0150]

[0151] Intermediate 3: 1-Ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate

[0152] [ka]

[0153] To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (2.00 g, 13.0 mmol) in DCM (40 mL) at 0 °C was added DMF (1-2 drops), followed by oxalyl chloride (3.50 mL, 40.8 mmol). The reaction mixture was stirred at room temperature for 2 h. After concentration in vacuum, the residue was dissolved in acetone (12 mL). The solution was added to a solution of potassium thiocyanate (1.64 g, 16.9 mmol) in acetone (28 mL) at 0 °C. The reaction mixture was stirred at room temperature for 1 h. After treatment with hexane (20 mL), the mixture was concentrated in vacuum. The impurities were solidified from hexane and DCM. The solid was filtered off and the filtrate was concentrated in vacuum. The residue was purified by column chromatography on SiO2 (hexane: EtOAc = 3: 1-2: 1) to provide the title compound (1.83 g, 72% between two steps) as a yellow oil. 1 H NMR (400MHz, CDCl3): δ6.72(1H,s), 4.49(2H,q,J=7.2Hz),2.28(3H,s),1.39(3H,t,J=7.2Hz).

[0154]

[0155] Intermediate 4: tert-Butyl 4-(chlorosulfonyl)piperazine-1-carboxylate

[0156] [ka]

[0157] To a solution of sulfonyl chloride (0.0520 mL, 0.644 mmol) in DCM (2.0 mL) at 0° C. was added a mixture of tert-butyl piperazine-1-carboxylate (0.100 g, 0.537 mmol) and pyridine (0.0650 mL, 0.805 mmol) in DCM (0.250 mL). The reaction mixture was stirred at room temperature for 1 h. After treatment with 1N aqueous HCl, the separated organic layer was washed with brine, dried over Na2SO4, and concentrated in vacuo to provide the title compound (54 mg, 35%). 1H NMR (400MHz, DMSO-d6): δ3.55 (4H, brs), 3.27 (4H, brs), 1.42 (9H, s).

[0158]

[0159] Intermediate 5: 4-Nitrophenylcyclopropylcarbamate

[0160] [ka]

[0161] To a solution of 4-nitrophenyl carbonochloridate (1.77 g, 8.76 mmol) in THF (25 mL) at room temperature, cyclopropanamine (0.610 mL, 8.76 mmol) was added followed by DIPEA (3.06 mL, 17.5 mmol). The reaction mixture was stirred at room temperature for 1 h. After quenching the reaction using water, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (hexane:EtOAc = 3:1) to provide the title compound (758 mg, 39%) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.27-8.23(3H,m),7.42-7.38(2H,m),2.60-2.56(1H,m),0.68-0.63(2H,m),0.54-0.50(2H,m).

[0162] Intermediate 6: 1-(tert-butyl) 4-(4-nitrophenyl)piperazine-1,4-dicarboxylate

[0163] [ka]

[0164] To a solution of 4-nitrophenyl chloroformate (0.541 g, 2.68 mmol) and TEA (0.561 mL, 4.03 mmol) in DCM (5.4 mL) was added tert-butyl piperazine-1-carboxylate (0.500 g, 2.68 mmol). The reaction mixture was stirred at room temperature for 3 h. After quenching the reaction using water, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (0.788 g, 84%) as a white solid. 1 H NMR (400MHz, DMSO-d6): δ8.28(2H,dd,J=6.8,2.4Hz),7.45(2H,dd,J=7.0,2.2Hz),3.59(1H,d,J=4.0Hz),3.41(6H,brs),1.42(9H,s).

[0165]

[0166] Intermediate 7: Cyclopropyl(4-nitrophenyl)carbonate

[0167] [ka]

[0168] A mixture of cyclopropanol (0.545 mL, 8.61 mmol) and pyridine (2.79 mL, 34.4 mmol) in DCM (86 mL) was stirred at room temperature for 10 min and cooled to 0 °C. After 4-nitrophenyl carbonochloridate (3.47 g, 17.2 mmol) was added, the reaction mixture was stirred at room temperature for 19 h. After concentration in vacuum, the residue was suspended in a mixture of EtOAc and hexanes (v / v = 1:1) and then stirred at room temperature for 10 min. The organic solvent was gently poured (repeated three times). The combined organic layers were concentrated in vacuum. The residue was purified by column chromatography on SiO2 (hexane only to hexane:EtOAc = 8:1) to provide the title compound (0.700 g, 36%) as a white solid. 1H NMR (400MHz, CDCl3): δ8.31-8.26(2H,m),7.41-7.37(2H m),4.31-4.26(1H,m),0.91-0.89(2H,m),0.84-0.82(2H,m).

[0169] Intermediate 8: tert-Butyl 4-(((4-nitrophenoxy)carbonyl)oxy)piperidine-1-carboxylate

[0170] [ka]

[0171] To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (0.100 g, 0.497 mmol) and TEA (0.104 mL, 0.745 mmol) in DCM (0.66 mL) was added 4-nitrophenyl carbonochloridate (0.110 g, 0.547 mmol). The reaction mixture was stirred at room temperature for 2 h. After concentration in vacuo, the residue was purified by column chromatography on SiO2 (hexane:EtOAc=4:1) to provide the title compound (98 mg, 54%). 1 H NMR(400MHz,DMSO-d6):δ8.32(2H,d,J=9.2Hz),7.58(2H,d,J=12.4Hz),4.91-4.87(1H,m),3 .63-3.57(2H, m), 3.22(2H, t, J=9.5Hz), 1.96-1.91(2H, m), 1.63-1.61(2H, m), 1.41(9H, s).

[0172]

[0173] Intermediate 9: Cyclopentyl(4-nitrophenyl)carbonate

[0174] [ka]

[0175] 4-Nitrophenyl chloroformate (2.34 g, 11.6 mmol) was added to a solution of cyclopentanol (0.527 mL, 5.80 mmol) in pyridine (1.8 mL) at 0 °C. The reaction mixture was stirred at room temperature for 3 h. The precipitated solid was suspended in DCM and filtered off. The filtrate was concentrated in vacuo. The residue was purified by column chromatography on SiO2 (hexane:EtOAc = 20:1 to 9:1) to provide the title compound (1.22 g, 84%) as a colorless oil. 1 H NMR (400MHz, DMSO-d6): δ8.30(2H,td,J=6.2,3.7Hz),7.55(2H,td,J=6.2,3.7Hz),5.16-5.13(1H,m),1.92-1.78(4H,m),1.72-1.58(4H,m).

[0176]

[0177] Intermediate 10: 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carbonate

[0178] [ka]

[0179] To a stirred solution of ethylene glycol (0.0900 mL, 1.61 mmol) in DCM (5.3 mL) at room temperature, pyridine (0.391 mL, 4.83 mmol) was added followed by TBS-Cl (0.292 mL, 1.69 mmol). The mixture was stirred at room temperature for 2 h. After 4-nitrophenyl-chloroformate (0.325 g, 1.61 mmol) was added, the reaction mixture was stirred at room temperature for 2 h. After concentration in vacuo, the residue was purified by column chromatography on SiO2 (hexane:EtOAc = 99:1 to 10:1) to provide the title compound (0.158 g, 28%) as a colorless oil. 1H NMR (400MHz, CDCl3): δ8.28(2H,dt,J=8.8,2.6Hz),7.38(2H,dt,J=8.4,2.8Hz),4.36(2H,t,J=4.8Hz),3.91(2H,t,J=4.8Hz),0.91(9H,s),0.10(6H,s).

[0180]

[0181] Intermediate 11: 4-Nitrophenyl 4-((tert-butyldimethylsilyl)oxy)piperidine-1-carboxylate

[0182] [ka]

[0183] Step A: 4-((tert-butyldimethylsilyl)oxy)piperidine

[0184] To a solution of piperidin-4-ol (0.197 mL, 1.98 mmol) in DCM (4.0 mL) at room temperature was added 1H-imidazole (0.538 g, 7.91 mmol) followed by TBS-Cl (0.715 g, 4.75 mmol). The reaction mixture was stirred at room temperature for 1 h. After quenching the reaction using water, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=100:10:1) to provide the title compound (0.436 g, quant.) as a colorless oil. 1 H NMR(400MHz,CDCl3):δ3.77-3.71(1H,m),3.09-3.03(2H,m),2.65-2.58(2H,m ),1.80-1.74(2H,m),1.43(2H,tt,J=12.9,4.6Hz),0.88(9H,s),0.05(6H,s).

[0185] Step B: 4-Nitrophenyl 4-((tert-butyldimethylsilyl)oxy)piperidine-1-carboxylate

[0186] To a solution of 4-((tert-butyldimethylsilyl)oxy)piperidine (0.426 g, 1.98 mmol) in DCM (6.6 mL) at room temperature was added pyridine (0.322 mL, 3.96 mmol) followed by 4-nitrophenyl carbonochloridate (0.399 g, 1.98 mmol). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by column chromatography on SiO2 (hexane:EtOAc = 10:1) to provide the title compound (0.169 g, 23% between two steps) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ8.25(2H,dt,J=8.2,2.8Hz),7.29(2H,dt,J=7.2,2.8Hz),4.03-3.99(1H,m),3.80-3.74 (1H,m),3.71-3.59(2H,m),3.58-3.52(1H,m),1.84-1.76(2H,m),1.65-1.59(2H,m),0.91(9H,s),0.08(6H,s).

[0187]

[0188] General Process for Alkynyloxy Intermediates

[0189] [ka]

[0190] Intermediate 12: tert-Butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0191] [ka]

[0192] Step A: tert-Butyl 3-formylazetidine-1-carboxylate

[0193] To a solution of tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (20.0 g, 107 mmol) in DCM (300 mL) at 0° C. was added Dess-Martin periodinane (90.6 g, 214 mmol). The reaction mixture was stirred at room temperature for 3 h. After adding 2M aqueous Na2S2O3 at 0° C., the mixture was stirred at 0° C. for another 30 min. After diluting with water, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 1:1) to provide the title compound (13.9 g, 42%) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ9.85 (1H, d, J=2.1Hz), 4.16-4.06 (4H, m), 1.44 (1H, s), 1.44 (9H, s). LC-MS:m / z=257.01[M-56] + .

[0194] Step B: tert-Butyl 3-ethynylazetidine-1-carboxylate

[0195] Dimethyl (1-diazo-2-oxopropyl)phosphonate (14.9 g, 83.6 mmol) was added to a mixture of tert-butyl 3-formylazetidine-1-carboxylate (12.9 g, 69.6 mmol) and K2CO3 (28.9 g, 209 mmol) in MeOH (150 mL) at 0 °C. The reaction mixture was stirred at room temperature overnight. After concentration in vacuum, the residue was dissolved in water and then extracted twice with Et2O. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography on SiO2 (petroleum ether: EtOAc = 10:1) to provide the title compound (8.80 g, 70%) as a yellow oil. 1 H NMR (400MHz, CDCl3): δ4.14(2H,t,J=8.6Hz),3.94(2H,dd,J=8.3,6.4Hz),3.30(1H,ttd,J=8.8,6.3,2.5Hz),2.28(1H,d,J=2.5Hz),1.44(9H,s). LC-MS:m / z=126.0[M-56]+ .

[0196] Step C: tert-Butyl 3-(3-hydroxyprop-1-yn-1-yl)azetidine-1-carboxylate

[0197] To a solution of tert-butyl 3-ethynylazetidine-1-carboxylate (8.10 g, 44.7 mmol) in THF (150 mL) at -78 °C was added LDA (2 M in THF, 14.4 g, 134 mmol) dropwise. The mixture was stirred at -78 °C for 30 min. After adding paraformaldehyde (21.1 g, 134 mmol) at -78 °C, the reaction mixture was stirred at room temperature for 1.5 h. After quenching the reaction using saturated aqueous NH4Cl solution, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:EtOAc = 2:1) to provide the title compound (5.37 g, 55%) as a yellow oil. 1 H NMR (400MHz, CDCl3): δ4.29(2H,d,J=2.0Hz),4.13(2H,t,J=8.5Hz),3.92(2H,dd,J=8.2,6.3Hz),3.34(1H,ttt,J=8.4,6.3,2.0Hz),1.44(9H,s). LC-MS:m / z=156.0[M-56] + .

[0198] Step D: tert-Butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0199] To a solution of methyl 4-chloro-3-hydroxy-5-nitrobenzoate (1.20 g, 5.21 mmol) in THF (23 mL) at 0° C. were successively added tert-butyl 3-(3-hydroxyprop-1-yn-1-yl)azetidine-1-carboxylate (1.00 g, 4.73 mmol), DIAD (1.10 mL, 5.68 mmol) and PPh3 (1.49 g, 5.68 mmol). The reaction mixture was stirred at room temperature overnight. After quenching the reaction with water, the mixture was extracted twice with EtOAc. The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (3.26 g, >99%) as a colorless oil. 1 H NMR(400MHz,DMSO-d6):δ8.14(1H,d,J=2.0Hz),8.00(1H,d,J=2.0Hz),5.20(2H,d,J=2.0Hz) ,4.09(2H,t,J=9.4Hz),3.92(3H,s),3.69(2H,t,J=6.8Hz),3.50-3.48(1H,m),1.36(9H,s).

[0200] Step E: tert-Butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0201] To a solution of tert-butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate (1.94 g, 4.57 mmol) at room temperature was added NH4OH (21.3 mL, 137 mmol). The reaction mixture was stirred at 50 °C for 3 h. After dilution with water, the mixture was extracted with DCM, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM only to DCM:MeOH = 98:2) to afford the title compound (0.977 g, 52% between two steps) as a yellow oil. 1H NMR(400MHz,DMSO-d6):δ8.27(1H,s),8.12(1H,d,J=2.0Hz),7.98(1H,d,J=1.6Hz),7.81(1H,s),5 .14(2H,d,J=1.6Hz),4.07(2H,t,J=8.4Hz),3.70(2H,t,J=6.8Hz),3.51-3.47(1H,m),1.36(9H,s).

[0202]

[0203] Intermediate 13: tert-Butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0204] [ka]

[0205] Step A: tert-3-ethynylpyrrolidine-1-carboxylate

[0206] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 3-formylpyrrolidine-1-carboxylate. The crude product was used in the next reaction without purification. 1 HNMR(400MHz,CDCl3):δ3.72-3.42(2H,m),3.30(2H,d,J=17.8Hz),2.94(1H, s),2.20-2.11(1H,m),2.11(1H,d,J=2.3Hz),2.01-1.87(1H,m),1.46(9H,s). LC-MS:m / z=140.0[M-56] + .

[0207] Step B: tert-3-(3-hydroxyprop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0208] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 3-ethynylpyrrolidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM:EtOAc=10:1) to provide the title compound (73%) as a yellow oil. 1 HNMR (400MHz, CDCl3): δ4.26(2H,d,J=2.0Hz),3.61(1H,dd,J=10.6,7.3Hz,1H),3.49(1H,ddd,J=12.2,7.9,4.6Hz), 3.39-3.20(2H,m),3.06-2.92(1H,m),2.13(1H,dtd,J=11.7,6.8,4.7Hz),1.91(1H,dq,J=12.3,8.0Hz),1.46(9H,s). LC-MS:m / z=170.0[M+H] + .

[0209] Step C: tert-Butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0210] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 3-(3-hydroxyprop-1-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (>99%) as a colorless oil. 1 H NMR(400MHz,DMSO-d6):δ8.11(1H,d,J=1.2Hz),7.98(1H,d,J=1.6Hz),5.13(2H,s), 3.91(3H,s),3.28-3.16(2H,m),3.10(2H,s),1.79-1.77(1H,m),1.38-1.35(11H,m).

[0211] Step D: tert-Butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0212] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to provide the title compound (37%) as a pale yellow solid. 1 H NMR(400MHz, DMSO-d:δ8.25(1H,s),8.11(1H,d,J=1.2Hz),7.98(1H,s),7.78(1H,s),5.09(2H,s),3.47(1H,d, J=4.4Hz), 3.20(1H,t,J=7.4Hz),3.11-3.08(2H,m),2.08-2.06(1H,m),1.81-1.75(1H,m),1.39-1.37(10H,m).

[0213]

[0214] Intermediate 14: tert-Butyl 3-(3-hydroxyprop-1-yn-1-yl)piperidine-1-carboxylate

[0215] [ka]

[0216] Step A: tert-Butyl 3-ethynylpiperidine-1-carboxylate

[0217] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 3-formylpiperidine-1-carboxylate. The crude product was used in the next reaction without purification. 1H NMR(400MHz,CDCl3):δ3.92(1H,d,J=13.8Hz),3.78-3.69(1H,m),3.07-2.93(2H,m),2.49-2.39(1H,m),2.06(1H,d, J=2.4Hz),2.03-1.93(1H,m),1.71(1H,dtt,J=13.8,5.1,3.4Hz),1.62-1.52(1H,m),1.46(9H,s),1.45-1.37(1H,m). LC-MS:m / z=154.0[M-56] + .

[0218] Step B: tert-Butyl 3-(3-hydroxyprop-1-yn-1-yl)piperidine-1-carboxylate

[0219] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 3-ethynylpiperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM:EtOAc=10:1) to provide the title compound (66%) as a yellow oil. 1 HNMR(400MHz,CDCl3)δ4.25(2H,d,J=2.0Hz),3.88(1H,ddt,J=13.2,4.0,1.3Hz),3.72(1H,dt,J=13.3,4.6Hz),3.05-2.92(2H, m),2.47(1H,ttt,J=9.4,3.7,1.9Hz),2.01-1.89(1H,m),1.75-1.67(1H,m),1.60-1.49(1H,m),1.46(9H,s),1.45-1.37(1H,m). LC-MS:m / z=184.0[M-56] + .

[0220] Step C: tert-Butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0221] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 3-(3-hydroxyprop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (>99%) as a colorless oil. 1 H NMR (400MHz, DMSO-d6): δ8.14(1H,d,J=2.0Hz),8.00(1H,d,J=1.2Hz),5.14(2H,d,J=1.2Hz),3.92 (3H,s),3.07(2H,s),2.55-2.50(2H,m),1.82-1.80(1H,m),1.57-1.48(2H,m),1.38-1.30(11H,m).

[0222] Step D: tert-Butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0223] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 3-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=9:1) to provide the title compound (68%) as a yellow oil. 1 H NMR(400MHz,DMSO-d6):δ8.26(1H,s),8.11(1H,d,J=2.0Hz),7.98(1H,d,J=1.6Hz),7.79(1H,s),5.08(2 H,d,J=2.0Hz),3.59(2H,s),3.04(2H,s),1.81(1H,s),1.59-1.47(2H,m),1.35(9H,s),1.34-1.29(2H,m)

[0224]

[0225] Intermediate 15: tert-Butyl 4-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0226] [ka]

[0227] Step A: tert-Butyl 4-(3-hydroxyprop-1-yn-1-yl)piperidine-1-carboxylate

[0228] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 4-ethynylpiperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 2:1) to provide the title compound (85%) as a yellow oil. 1 HNMR(400MHz,DMSO-d6):δ4.34-4.24(2H,m),3.71(2H,ddd,J=13.4,6.5,3.8Hz),3.14(2H,ddd,J=13.5,8.7,3.4Hz),2.6 0(1H,dddt,J=8.2,6.0,3.9,1.9Hz),1.77(2H,ddt,J=13.6,7.1,3.7Hz),1.56(2H,dtd,J=12.7,8.6,3.8Hz),1.45(9H,s). LC-MS:m / z=240.3[M+H] + .

[0229] Step B: tert-Butyl 4-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0230] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 4-(3-hydroxyprop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (47%) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ8.06(1H,d,J=1.2Hz),7.96(1H,d,J=1.6Hz),4.91(2H,d,J=1.6Hz),3.96(3H,s) ,3.61-3.59(2H,m),3.20-3.13(2H,m),2.62(1H,m),1.76-1.72(2H,m),1.57-1.49(2H,m),1.44(9H,s).

[0231] Step C: tert-Butyl 4-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0232] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using tert-butyl 4-(3-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to provide the title compound (32%) as a white solid. 1 H NMR (400MHz, CDCl3): δ8.25(1H,s),8.11(1H,d,J=1.2Hz),8.01(1H,d,J=2.0Hz),7.78(1H,s),5.10( 2H, d, J=1.2Hz), 3.46(2H, m), 3.09-3.05(2H, m), 2.76-2.64(1H, m), 1.70-1.65(2H, m), 1.38(11H, s).

[0233]

[0234] Intermediate 16: Methyl 4-chloro-3-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-5-nitrobenzamide

[0235] [ka]

[0236] Step A: 5-chloro-2-methylpent-3-yn-2-ol

[0237] To a solution of 3-chloroprop-1-yne (9.2 M in toluene, 4.38 mL, 40.3 mmol) in dry THF (134 mL) at -78 °C, n-BuLi (2.5 M in hexane, 16.1 mL, 40.3 mmol) was added dropwise. The mixture was stirred at -78 °C for 30 min. After adding dry acetone (2.96 mL, 40.3 mmol) over 5 min, the reaction mixture was stirred at -78 °C for 3 h and warmed to 0 °C. After quenching the reaction using 1 M aqueous NH4Cl solution, the mixture was extracted twice with Et2O. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (hexane: EtOAc = 4:1) to provide the title compound (4.39 g, 82%) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ4.12(2H,s), 2.78(1H,s), 1.48(6H,s).

[0238] Step B: Methyl 4-chloro-3-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-5-nitrobenzoate

[0239] A mixture of methyl 4-chloro-3-hydroxy-5-nitrobenzoate (500 mg, 2.16 mmol) and K2CO3 (358 mg, 2.59 mmol) in DMF (2.2 mL) was stirred at room temperature for 30 min. After adding 5-chloro-2-methylpent-3-yn-2-ol (315 mg, 2.38 mmol) at room temperature, the reaction mixture was stirred at 70 °C overnight. After filtration through a Celite pad with EtOAc washing, the filtrate was washed with water and brine, dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography on SiO2 (hexane:EtOAc = 3:1 to 1:1) to provide the title compound (578 mg, 82%) as a yellow oil. 1 H NMR (400MHz, CDCl3): 7.98(1H,d,J=1.2Hz),7.91(1H,d,J=1.6Hz),4.88(2H,s),3.92(3H,s),2.84(1H,s),1.45(6H,s).

[0240] Step C: Methyl 4-chloro-3-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-5-nitrobenzamide

[0241] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using methyl 4-chloro-3-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-5-nitrobenzoate (1.05 g, 3.20 mmol). The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to provide the title compound (62%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.26(1H,s),8.10(1H,d,J=2.0Hz),7.97(1H,d,J=1.2Hz),7.80(1H,s),5.44(1H,s),5.10(2H,s),1.33(6H,s).

[0242]

[0243] Intermediate 17: 2-(Dimethylamino)ethyl (4-nitrophenyl) carbonate hydrochloride

[0244] [ka]

[0245] To a solution of 2-(dimethylamino)ethan-1-ol (0.562 mL, 5.61 mmol) in THF (19 mL) at 0° C. was added 4-nitrophenyl carbonochloridate (1.13 g, 5.61 mmol). The reaction mixture was stirred at room temperature for 3 h. After treatment with EtOAc, the precipitated solid was collected by filtration, washed with EtOAc, and dried under vacuum to provide the title compound (1.34 g, 82%) as a white solid. 1 H NMR (400MHz, DMSO-d6): δ8.13-8.09(2H,m),6.98-6.94(2H,m),5.33(1H,brs),3.72(2H,s),3.10-3.13(2H,m),2.76(6H,s).

[0246]

[0247] Intermediate 18: tert-Butyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0248] [ka]

[0249] Step A: tert-Butyl 4-(4-hydroxybut-2-yn-1-yl)piperazine-1-carboxylate

[0250] To a solution of tert-butyl piperazine-1-carboxylate (6.20 g, 33.3 mmol) and K2CO3 (4.60 g, 33.3 mmol) in DMF (80 mL) at room temperature was added 4-[(4-methylbenzenesulfonyl)oxy]but-2-yn-1-ol (4.00 g, 16.6 mmol). The reaction mixture was stirred at room temperature for 2 h. After partitioning between EtOAc and water, the separated aqueous layer was extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 1:1) to provide the title compound (2.30 g, 54%) as a pale yellow oil. 1 HNMR (400MHz, CDCl3): δ4.28(2H,t,J=1.9Hz),3.47(4H,t,J=5.1Hz),3.33(2H,t,J=2.0Hz),2.99(1H,s),2.51(4H,t,J=5.1Hz),1.46(9H,s). LC-MS:m / z=255[M+H] + .

[0251] Step B: tert-Butyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0252] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 4-(4-hydroxybut-2-yn-1-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (Hexane:EtOAc=1 / 1 to DCM:MeOH=98 / 2) to provide the title compound (quantitative) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ8.07(1H,d,J=1.2Hz),7.92(1H,d,J=1.2Hz),4.94(2H,t,J=1.6H z),3.97(3H,s),3.43(4H,t,J=4.8Hz),3.34(2H,t,J=1.6Hz),2.45(4H,s),1.46(9H,s). LC-MS:m / z=468.1[M+H]+ .

[0253] Step C: tert-Butyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0254] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=9:1) to provide the title compound (36%) as a yellow solid. 1H NMR (400MHz, DMSO-d6): δ 8.26 (1H,s), 8.11 (1H,d,J=1.6Hz), 7.99 (1H,d,J=2.0Hz), 7.79 (1H,s), 5.15 (2H,s), 3.29-3.26 (4H,m), 2.32 (4H,t,J=4.4Hz), 1.38 (9H,s). LC-MS: m / z=453.1 [M+H] + .

[0255]

[0256] Intermediate 19: Isopropyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0257] [ka]

[0258] Step A: Isopropyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0259] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using isopropyl 4-(4-hydroxybut-2-yn-1-yl)piperazine-1-carboxylate and methyl 4-chloro-3-hydroxy-5-nitrobenzoate. The crude product was purified by column chromatography on NH-SiO2 (Hexanes:EtOAc=2:1 to 1:1) followed by SiO2 (Hexanes:EtOAc=1:1 to EtOAc:MeOH=10:1) to provide the title compound (93%) as a yellow oil. 1 H NMR (400MHz, CDCl3): δ8.06(1H,d,J=1.6Hz),7.91(1H,d,J=1.6Hz),4.94(2H,t,J=2.0Hz),4.92-4.86 (1H,m),3.97(3H,s),3.48-3.46(4H,m),3.35(2H,t,J=2.0Hz),2.45(4H,brs),1.24(6H,d,J=6.0Hz).

[0260] Step B: Isopropyl 4-(4-(5-carbamoyl-2-chloro-3-isopropylphenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0261] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using isopropyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:EtOAc=7:3 to DCM:MeOH=20:1) to provide the title compound (50%) as a pale yellow solid. 1 H NMR (400MHz, CDCl3): δ8.42(1H,s),8.13(1H,s),7.59(1H,s),7.43(1H,s),5.74(2H,brs),4.89(1H,t,J=6.0Hz), 4.78 (2H, s), 4.26 (3H, s), 3.91 (2H, s), 3.85 (2H, s), 3.44 (4H, s), 3.34 (2H, s), 2.40 (4H, s), 1.23 (6H, d, J=6.0Hz).

[0262]

[0263] Intermediate 20: 4-Chloro-3-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-5-nitrobenzamide

[0264] [ka]

[0265] Step A: 2-Methyl-1-(piperazin-1-yl)propan-1-one

[0266] To a solution of piperazine (20.2 g, 235 mmol) in AcOH (50 mL) at room temperature was added isobutyryl chloride (5.00 g, 46.9 mmol) slowly. The reaction mixture was stirred at room temperature for 2 h. 1N aqueous NaOH was used to basify to pH 9, and the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 9:1) to provide the title compound (6.00 g, 82%) as a colorless oil. LC-MS: m / z = 157 [M + H] + .

[0267] Step B: 1-[4-(4-hydroxybut-2-yn-1-yl)piperazin-1-yl]-2-methylpropan-1-one

[0268] The title compound was prepared in a similar manner to Intermediate 18 (Step A) using 2-methyl-1-(piperazin-1-yl)propan-1-one and 4-[(4-methylbenzenesulfonyl)oxy]but-2-yn-1-ol. The crude product was purified by column chromatography on SiO2 (EtOAc:MeOH=10:1) to provide the title compound (15%) as a pale yellow oil. 1H NMR (400MHz, CDCl3): δ4.30(2H,d,J=1.9Hz),3.68(2H,q,J=14.9,10.1Hz),3.57(2H,t,J=5.1Hz),3.37(2H ,d,J=2.0Hz),2.80(1H,hept,J=6.7Hz),2.57(4H,dt,J=11.1,4.9Hz),2.23(1H,s),1.14(6H,d,J=6.7Hz). LC-MS:m / z=225[M+H] + .

[0269] Step C: Methyl 4-chloro-3-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-5-nitrobenzoate

[0270] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and 1-(4-(4-hydroxybut-2-yn-1-yl)piperazin-1-yl)-2-methylpropan-1-one. The crude product was purified by column chromatography on SiO2 (hexanes:EtOAc=3:1) to provide the title compound (>99%) as a colorless oil. 1 H NMR (400MHz, CDCl3): δ8.14(1H,d,J=1.6Hz),7.99(1H,d,J=2.0Hz),5.22(2H,s),3. 92(3H,s),3.40(6H,m),2.84-2.77(1H,m),2.35-2.34(4H,m),0.95(6H,d,J=6.8Hz). LC-MS:m / z=438.1[M+H] + .

[0271] Step D: 4-Chloro-3-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-5-nitrobenzamide

[0272] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using methyl 4-chloro-3-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-5-nitrobenzoate. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=98:2) to provide the title compound (29% between two steps) as a yellow solid. 1 H NMR(400MHz,DMSO-d6):δ8.25(1H,s),8.11(1H,d,J=2.0Hz),7.99(1H,d,J=2.0Hz),7.79(1H,s),5.15(2 H,s),3.42(4H,s),3.36(2H,s),2.82(1H,t,J=7.2Hz),2.38(2H,s),2.33(2H,s),0.96(6H,d,J=6.4Hz). LC-MS:m / z=423.1[M+H] + .

[0273]

[0274] Intermediate 21: 4-chloro-3-((4-morpholinobut-2-yn-1-yl)oxy)-5-nitrobenzamide

[0275] [ka]

[0276] Step A: 4-(morpholin-4-yl)but-2-yn-1-ol

[0277] The title compound was prepared in a similar manner to Intermediate 18 (Step A) using morpholine and 4-[(4-methylbenzenesulfonyl)oxy]but-2-yn-1-ol. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 9:1) to provide the title compound (93%) as a pale yellow oil. 1HNMR (400MHz, CDCl3): δ4.28(2H,t,J=2.0Hz),3.78-3.71(4H,m),3.67(1H,s),3.31(2H,t,J=2.0Hz),2.58(4H,dd,J=5.7,3.8Hz). LC-MS:m / z=156.0[M+H] + .

[0278]

[0279] Step B: Methyl 4-chloro-3-((4-morpholinobut-2-yn-1-yl)oxy)-5-nitrobenzoate

[0280] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and 4-morpholinobut-2-yn-1-ol. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (quantitative) as a colorless oil. 1 H NMR(400MHz,DMSO-d6):δ8.15(1H,d,J=1.6Hz),8.01(1H,d,J=2.0Hz),5.23(2H,s) ,3.91(3H,s),3.51(4H,t,J=4.4Hz),3.33(2H,d,J=1.6Hz),2.34(4H,t,J=4.8Hz).

[0281] Step C: 4-chloro-3-((4-morpholinobut-2-yn-1-yl)oxy)-5-nitrobenzamide

[0282] The title compound was prepared using methyl 4-chloro-3-((4-morpholinobut-2-yn-1-yl)oxy)-5-nitrobenzoate in a similar manner to Intermediate 12 (Step E). The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to provide the title compound (60%) as a yellow solid. 1H NMR(400MHz,DMSO-d6):δ8.25(1H,s),8.11(1H,d,J=2.0Hz),8.01(1H,d,J=2.0Hz),7.80 (1H,s),5.16(2H,s),3.52(4H,t,J=4.8Hz),3.31(2H,d,J=2.0Hz),2.35(4H,t,J=4.8Hz).

[0283]

[0284] Intermediate 22: tert-Butyl 4-(5-(5-carbamoyl-2-chloro-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0285] [ka]

[0286] Step A: tert-4-(2-methylbut-3-yn-2-yl)piperazine-1-carboxylate

[0287] To a mixture of tert-butyl piperazine-1-carboxylate (6.00 g, 32.2 mmol), CuCl (0.320 g, 3.22 mmol) and TEA (6.52 g, 64.4 mmol) in THF (60 mL) at 0 °C was added 3-chloro-3-methylbut-1-yne (3.63 g, 35.4 mmol). The reaction mixture was stirred at room temperature for 5 h. After quenching the reaction using ice water, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether: EtOAc = 6:1) to provide the title compound (7.00 g, 86%) as a colorless oil. 1 HNMR (400MHz, DMSO-d6): δ3.31(4H,t,J=5.0Hz),3.18(1H,s),2.45(4H,t,J=5.1Hz),1.39(9H,s),1.30(6H,s).

[0288] Step B: tert-Butyl 4-(5-hydroxy-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0289] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 4-(2-methylbut-3-yn-2-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM:EtOAc=1:1) to provide the title compound (31%) as a pale yellow oil. 1 HNMR (400MHz, DMSO-d6): δ5.08(1H,t,J=5.9Hz),4.07(2H,d,J=5.9Hz),3.30(4H,d,J=4.9Hz),2.47(4H,t,J=5.1Hz),1.40(9H,s),1.29(6H,s). LC-MS:m / z=283[M+H] + .

[0290] Step C: tert-Butyl 4-(5-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0291] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 4-(5-hydroxy-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (quantitative) as a colorless oil. LC-MS: m / z=496.1 [M+H] + .

[0292] Step D: tert-Butyl 4-(5-(5-carbamoyl-2-chloro-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0293] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 4-(5-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to afford the title compound (30% between two steps) as a pale yellow solid. 1H NMR(400MHz,DMSO-d6):δ8.24(1H,s),8.11(1H,d,J=1.6Hz),8.03(1H,d,J=2.0Hz),7. 76(1H,s),5.13(2H,s),3.24(4H,t,J=4.6Hz),2.35(4H,s),1.38(9H,s),1.25(6H,s).

[0294]

[0295] Intermediate 23: tert-Butyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-yl)piperidine-1-carboxylate

[0296] [ka]

[0297] Step A: tert-Butyl 4-(prop-2-yn-1-yl)piperidine-1-carboxylate

[0298] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 4-(2-oxoethyl)piperidine-1-carboxylate and dimethyl (1-diazo-2-oxopropyl)phosphonate. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 3:1) to provide the title compound (98%) as a colorless oil. 1H NMR(400MHz,DMSO-d6):δ3.93(2H,d,J=13.1Hz),2.80(1H,t,J=2.7Hz),2.68(2H,s),2.13(2H,dd,J= 6.6,2.7Hz),1.71-1.63(2H,m),1.63-1.51(1H,m),1.39(9H,s),1.07(2H,tdd,J=12.9,11.5,4.4Hz).

[0299] Step B: tert-Butyl 4-(4-hydroxybut-2-yn-1-yl)piperidine-1-carboxylate

[0300] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 4-(prop-2-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM:EtOAc=20:1) to provide the title compound (73%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6): δ5.03(1H,t,J=5.9Hz),4.03(2H,dt,J=5.9,2.2Hz),3.93(2H,d,J=13.1Hz),2.67(2H,d,J=4.9Hz), 2.15(2H,dt,J=6.7,2.2Hz),1.72-1.63(2H,m),1.55(1H,dddd,J=13.9,11.3,6.7,3.1Hz),1.41(9H,s),1.13-0.98(2H,m).

[0301] Step C: tert-Butyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0302] The title compound was prepared using tert-butyl 4-(4-hydroxybut-2-yn-1-yl)piperidine-1-carboxylate and methyl 4-chloro-3-hydroxy-5-nitrobenzoate in a similar manner to intermediate 12 (Step D). The crude product was purified by column chromatography on NH-SiO2 (hexane:EtOAc=2:1 to 1:1) to provide the title compound as a yellow oil.1 H NMR(400MHz,CDCl3);δ8.06(1H,d,J=1.6Hz),7.93(1H,d,J=1.6Hz),5.00-4.94(1H,m),4.90(2H,t,J=2.0Hz),4.0 8(2H,brs),3.97(3H,s),2.67-2.60(2H,m),2.18-2.16(2H,m),1.70-1.67(2H,m),1.44(9H,s),1.14-1.04(2H,m).

[0303] Step D: tert-Butyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-yl)piperidine-1-carboxylate

[0304] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:EtOAc=7:3 to DCM:MeOH=20:1) to provide the title compound (51% between two steps) as a pale yellow solid. 1 H NMR (400 MHz, CDCl3); 1 H NMR (400MHz, CDCl3): δ7.86(1H,d,J=1.6Hz),7.75(1H,d,J=1.6Hz),5.00-4.94(1H,m),4.92(2H,t,J=2.0Hz ),4.05(2H,brs),2.67-2.60(2H,m),2.20-2.19(2H,m),1.65-1.61(2H,m),1.44(9H,s),1.14-1.04(2H,m).

[0305]

[0306] Intermediate 24: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0307] [ka]

[0308] Step A: tert-Butyl 3-(2-hydroxyethyl)azetidine-1-carboxylate

[0309] To a solution of tert-butyl 3-(2-methoxy-2-oxoethyl)azetidine-1-carboxylate (25.0 g, 109 mmol) in THF (250 mL) at 0° C. was added LiBH4 (191 mL, 382 mmol) in portions. The reaction mixture was stirred at room temperature overnight. After quenching the reaction with water, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (20.0 g, 91%) as a colorless oil. 1H NMR (400MHz, CDCl3): δ4.02(2H,q,J=8.6Hz),3.76(1H,dt,J=19.7,6.2Hz),3.64-3.57(3H,m),2.71-2.56(1H,m),1.88-1.80(2H,m),1.43(9H,s). LC-MS:m / z=202[M+H] + .

[0310] Step B: tert-Butyl 3-(2-oxoethyl)azetidine-1-carboxylate

[0311] A mixture of tert-butyl 3-(2-hydroxyethyl)azetidine-1-carboxylate (20.0 g, 99.4 mmol) and Dess-Martin periodinane (84.3 g, 199 mmol) in DCM (200 mL) was stirred at room temperature for 2 h. After saturated aqueous NaHCO3 was added, the mixture was filtered and the filter cake was washed with DCM. The filtrate was concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 3:1) to provide the title compound (10.0 g, 50%) as a yellow oil. 1HNMR (400MHz, CDCl3): δ9.77(1H,d,J=0.9Hz),3.74-3.42(4H,m),2.85-2.77(2H,m),2.07(1H,s),1.43(9H,s). LC-MS:m / z=200[M+H] + .

[0312] Step C: tert-Butyl 3-(prop-2-yn-1-yl)azetidine-1-carboxylate

[0313] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 3-(2-oxoethyl)azetidine-1-carboxylate and dimethyl (1-diazo-2-oxopropyl)phosphonate. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 19:1) to provide the title compound (60%) as a yellow oil. 1 HNMR (400MHz, CDCl3): δ4.06-3.97(2H,m),3.69(2H,dd,J=8.8,5.3Hz),2.44(2H,dd,J=6.9,2.7Hz),1.98(1H,t,J=2.6Hz),1.44(9H,s). LC-MS:m / z=196[M+H] + .

[0314] Step D: tert-Butyl 3-(4-hydroxybut-2-yn-1-yl)azetidine-1-carboxylate

[0315] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 3-(prop-2-yn-1-yl)azetidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 7:3) to provide the title compound (31%) as a yellow oil. 1HNMR(400MHz, CDCl3): δ4.24(2H,t,J=2.1Hz),4.02(2H,t,J=8.4Hz),3.68(2H, dd,J=8.8,5.2Hz),2.74-2.63(1H,m),2.47(2H,dt,J=6.8,2.2Hz),1.44(9H,s). LC-MS:m / z=226[M+H] + .

[0316] Step E: tert-Butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0317] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 3-(4-hydroxybut-2-yn-1-yl)azetidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (crude) as a colorless oil. LC-MS: m / z=339.0 [M-99] + .

[0318] Step F: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0319] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=9:1) to provide the title compound (26% between two steps) as a yellow oil. 1H NMR (400MHz, DMSO-d6): δ8.25(1H,s),8.11(1H,d,J=1.6Hz),7.97(1H,d,J=1.6Hz),7.78(1H,s),5.0 8(2H,s),3.83(2H,t,J=8.4Hz),3.46(2H,t,J=6.6Hz),2.67-2.60(1H,m),2.52(2H,s),1.33(9H,s). LC-MS:m / z=324.0[M-99] + .

[0320]

[0321] Intermediate 25: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0322] [ka]

[0323] Step A: tert-Butyl 3-[(E)-2-methoxyethenyl]pyrrolidine-1-carboxylate

[0324] KO in THF (180 mL) at room temperature t-To a mixture of Bu (20.3 g, 181 mmol) was added (methoxymethyl)triphenyl-lambda 5-phosphane chloride (62.1 g, 181 mmol). The mixture was stirred at room temperature for 30 min. tert-Butyl 3-formylpyrrolidine-1-carboxylate (18.0 g, 90.3 mmol) was added portionwise at room temperature, and then the reaction mixture was stirred at room temperature overnight. After quenching the reaction with water, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (petroleum ether:EtOAc = 3:1) to provide the title compound (6.30 g, 31%) as a yellow liquid. 1 HNMR(400MHz,CDCl3):δ6.38(1H,d,J=12.6Hz),4.64(1H,dd,J=12.7,8.5Hz),3.60(1H,s),3.52(2H,s),3.27 (1H,s),2.93(1H,s),2.68(1H,d,J=8.3Hz),1.98(1H,dddq,J=15.5,9.4,6.6,2.9Hz),1.46(9H,d,J=1.7Hz). LC-MS:m / z=228[M+H] + .

[0325] Step B: tert-Butyl 3-(2-oxoethyl)pyrrolidine-1-carboxylate

[0326] A mixture of tert-butyl 3-[(E)-2-methoxyethenyl]pyrrolidine-1-carboxylate (7.10 g, 31.2 mmol) and formic acid (28 mL) was stirred at room temperature for 40 min. After neutralization with saturated aqueous NaHCO3, the mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (5.00 g, 75%) as a yellow oil. 1HNMR (400MHz, CDCl3): δ9.79 (1H t,J=1.3Hz),3.63(1H,dd,J=10.8,7.0Hz),3.45(1H,ddd,J=11.5,8.2,3.8Hz),3.34-3.26(1H,m),2.92(1H, dd,J=10.8,7.6Hz),2.68-2.51(3H,m),2.10(1H,dtd,J=10.2,6.6,3.8Hz),1.61-1.49(1H,m),1.46(9H,s). LC-MS:m / z=214[M+H] + .

[0327] Step C: tert-Butyl 3-(prop-2-yn-1-yl)pyrrolidine-1-carboxylate

[0328] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 3-(2-hydroxyethyl)pyrrolidine-1-carboxylate and dimethyl (1-diazo-2-oxopropyl)phosphonate. The crude product was used in the next reaction without purification as a yellow oil. 1 HNMR(400MHz,CDCl3):δ3.47(1H,dd,J=10.9,7.1Hz),3.39(1H,ddd,J=15.1,7.4,3.4Hz),3.24(1H,dt,J=10.9,7.7Hz),2.99(1H,dd,J =10.8,7.4Hz),2.30(1H,dt,J=14.2,7.1Hz),2.21(2H,dt,J=6.9,2.5Hz),2.00-1.90(2H,m),1.68-1.59(1H,m),1.39(9H,d,J=0.9Hz). LC-MS:m / z=210[M+H] + .

[0329] Step D: tert-Butyl 3-(4-hydroxybut-2-yn-1-yl)pyrrolidine-1-carboxylate

[0330] The title compound was prepared using tert-butyl 3-(prop-2-yn-1-yl)pyrrolidine-1-carboxylate in a similar manner to Intermediate 12 (Step C). The crude product was purified by column chromatography on SiO2 (petroleum ether: EtOAc = 7:3) to afford the title compound (50% between two steps) as a yellow oil. 1 HNMR(400MHz,CDCl3):δ4.25(2H,t,J=2.0Hz),3.56-3.41(2H,m),3.30(1H,dt,J=10.9,7.8Hz),3.07(1H,dd,J=1 0.8,6.8Hz),2.43-2.30(2H,m),2.34-2.27(1H,m),2.07-1.95(2H,m),1.69(1H,dq,J=12.7,8.0Hz),1.46(9H,s). LC-MS:m / z=240[M+H] + .

[0331] Step E: tert-Butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0332] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 3-(4-hydroxybut-2-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (hexane:EtOAc=3:1) to provide the title compound (quantitative) as a colorless oil. 1 H NMR(400MHz,DMSO-d6):δ8.13(1H,d,J=1.6Hz),7.99(1H,d,J=2.0Hz),5.16(2H,s),3.91(3H,s),3.26-3.23(1H,m),3.17-3.09(1H,m) ,2.81(1H,dd,J=10.6,8.2Hz),2.36(2H,d,J=6.0Hz),2.29-2.20(1H,m),1.91-1.82(1H,m),1.57-1.47(1H,m),1.36(9H,d,J=7.6Hz).

[0333] Step F: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0334] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=95:5) to provide the title compound (73%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6):δ8.24(1H,s),8.11(1H,s),7.99(1H,s),7.78(1H,s),5.09(2H,s),3.36-3.33(1H,m),3.30-3.25(1H,m),3.17-3.09 (1H,m),2.83(1H,dd,J=10.6,7.8Hz),2.36(2H,d,J=6.4Hz),2.28-2.24(1H,m),1.88-1.86(1H,m),1.56-1.49(1H,m),1.37(9H,d,J=6.4Hz).

[0335]

[0336] Intermediate 26: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0337] [ka]

[0338] Step A: tert-Butyl 3-[(E)-2-methoxyethenyl]piperidine-1-carboxylate

[0339] The title compound was prepared in a similar manner to Intermediate 25 (Step A) using tert-butyl 3-formylpiperidine-1-carboxylate and (methoxymethyl)triphenyl-lambda 5-phosphane chloride. The crude product was purified by column chromatography on SiO2 (petroleum ether: EtOAc = 9:1) to provide the title compound (63%) as a yellow oil. LC-MS: m / z = 242 [M + H] + .

[0340] Step B: tert-Butyl 3-(2-oxoethyl)piperidine-1-carboxylate

[0341] The title compound was prepared in a similar manner to Intermediate 25 (Step B) using tert-butyl 3-[(E)-2-methoxyethenyl]piperidine-1-carboxylate. The crude product was used in the next reaction without purification as a yellow oil. 1 HNMR(400MHz, CDCl3): δ9.78(1H,t,J=1.9Hz),3.91-3.78(2H,m),2.89(1H,ddd,J=13.5,10.6,3.3Hz),2.67(1H,t,J=11. 3Hz),2.41(1H,ddd,J=16.9,6.6,1.7Hz),2.30(1H,ddd,J=16.9,7.1,2.1Hz),2.10(1H,ttt,J=10.4,7.0,3.8Hz),1.85(1H ddt,J=13.2,5.3,3.8Hz),1.64(1H,dq,J=12.9,4.1Hz),1.55-1.48(1H,m),1.46(9H,s),1.20(1H,dtd,J=14.0,10.4,4.0Hz). LC-MS:m / z=228[M+H] + .

[0342] Step C: tert-Butyl 3-(prop-2-yn-1-yl)piperidine-1-carboxylate

[0343] The title compound was prepared in a similar manner to Intermediate 12 (Step B) using tert-butyl 3-(2-oxoethyl)piperidine-1-carboxylate and dimethyl (1-diazo-2-oxopropyl)phosphonate. The crude product was used in the next reaction without purification as a yellow oil. 1 HNMR(400MHz,CDCl3):δ4.05-3.98(1H,m),3.89(1H,dtt,J=13.2,4.1,1.4Hz), 2.79(1H,ddd,J=13.2,11.2,3.2Hz),2.61(1H,dd,J=13.1,10.1Hz),2.17-2.11( 2H,m),1.99(1H,t,J=2.7Hz),1.88(1H,dt,J=13.1,3.9Hz),1.68(3H,dddt,J=1 8.0,14.2,8.1,4.1Hz),1.46(9H,s),1.25(1H,dddd,J=14.8,13.1,8.5,3.3Hz). LC-MS: m / z=224[M+H] + .

[0344] Step D: tert-Butyl 3-(4-hydroxybut-2-yn-1-yl)piperidine-1-carboxylate

[0345] The title compound was prepared in a similar manner to Intermediate 12 (Step C) using tert-butyl 3-(prop-2-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (petroleum ether:EtOAc=4:1) to provide the title compound (16% over three steps) as a yellow oil. 1HNMR(400MHz, CDCl3): δ4.25(2H,t,J=2.2Hz),4.05-3.96(1H,m),3.93-3.78(1H ,m),2.79(1H,ddd,J=13.1,11.1,3.2Hz),2.60(1H,dd,J=13.1,10.1Hz),2.17(2H ,dt,J=6.8,2.2Hz),1.87(2H,d,J=17.3Hz),1.66(2H,dtt,J=15.0,7.5,3.7Hz), 1.46(9H,s),1.43-1.32(1H,m),1.32-1.15(1H,m),0.92(1H,dt,J=14.2,7.2Hz). LC-MS: m / z=254[M+H] + .

[0346] Step E: tert-Butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0347] The title compound was prepared in a similar manner to Intermediate 12 (Step D) using tert-butyl 3-(4-hydroxybut-2-yn-1-yl)piperidine-1-carboxylate and methyl 4-chloro-3-hydroxy-5-nitrobenzoate. The crude product was purified by column chromatography on NH-SiO2 (hexane:EtOAc=3:1) to provide the title compound as a colorless oil. 1 H NMR(400MHz,DMSO-d6):δ8.13(1H,d,J=1.6Hz),7.99(1H,d,J=2.0Hz),5.14(2H,s),3.90(3H,s),3.73-3.70(2 H,m),2.62(1H,s),2.19(2H,d,J=5.2Hz),1.69(1H,d,J=12.8Hz),1.52-1.48(2H,m),1.38(1H,s),1.35(9H,s). LC-MS:m / z=367.1[M-99] + .

[0348] Step F: tert-Butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0349] The title compound was prepared in a similar manner to Intermediate 12 (Step E) using tert-butyl 3-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=9:1) to provide the title compound (34% between two steps) as a yellow foam. 1H NMR (400MHz, DMSO-d6): δ8.24(1H,s),8.10(1H,d,J=2.0Hz),7.99(1H,d,J=1.2Hz),7.78(1H,s),5.10(2H,s),3.80-3. 73(2H,m),2.68-2.63(1H,m),2.21-2.18(2H,m),1.71-1.68(1H,m),1.54-1.46(2H,m),1.38(9H,s),1.27-1.24(1H,m). LC-MS:m / z=352.0[M-99] + .

[0350]

[0351] [ka]

[0352] Working Example Example 1: (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0353] [ka]

[0354] Step A: tert-Butyl (E)-4-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0355] To a solution of tert-butyl 4-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (Intermediate 15, 0.388 g, 0.886 mmol) in DMSO (4.4 mL) was added DIPEA (0.774 mL, 4.43 mmol) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1, 0.588 g, 1.77 mmol). The reaction mixture was stirred at 120° C. for 3 h. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on NH-SiO2 (DCM only to DCM:MeOH=98:2) to afford the title compound (0.265 g, 43%) as an orange solid. 1 H NMR (400MHz, DMSO-d6): δ8.19(1H,d,J=2.0Hz),8.10(1H,d,J=1.6Hz),7.96(2H,m),7. 77(1H,t,J=6.4Hz),7.61(1H,d,J=2.0Hz),7.35(1H,d,J=1.6Hz),7.30(1H,brs),5.59( 2H,t,J=4.4Hz),4.78(2H,d,J=1.6Hz),4.10(4H,m),3.83(3H,s),3.80(3H,s),3.47-3 .42(2H,m),3.07(2H,t,J=9.6Hz),2.66(1H,m),1.68-1.63(2H,m),1.38-1.37(11H,m).

[0356] Step B: tert-Butyl (E)-4-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)-amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0357] To a solution of tert-butyl (E)-4-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitro-phenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (0.265 g, 0.380 mmol) in a mixture of MeOH (7.5 mL) and THF (5.8 mL) at 0 °C was added a solution of sodium hyposulfite (0.927 g, 5.32 mmol) in water (5.8 mL). The reaction mixture was stirred at room temperature for 1 h. After quenching the reaction using saturated aqueous NaHCO3 solution, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (0.195 g, 81%) as a light brown solid. LC-MS: m / z=637.2[M+H] + .

[0358] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0359] To a solution of tert-butyl (E)-4-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)-phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (0.195 g, 0.306 mmol) in DMF (2.5 mL) at room temperature was added cyanyl bromide (81.0 mg, 0.766 mmol). The reaction mixture was stirred at room temperature overnight and then heated at 70° C. for 1 h. After concentration in vacuo, the residue was purified by recrystallization from Et2O / MeOH. The solid was collected by filtration, washed with Et2O and dried under vacuum to provide the title compound (0.230 g, 89%) as a yellow solid. LC-MS: m / z=687.2 [M+H] + .

[0360] Step D: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0361] To a solution of methyl (E)-2-(bromo-15-azanyl)-1-(4-(2-(bromo-15-azanyl)-7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate (0.230 g, 0.271 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (92.0 mg, 0.596 mmol) in DMF (2.7 mL) at room temperature was added HATU (0.227 g, 0.596 mmol) and DIPEA (0.284 mL, 1.63 mmol). The reaction mixture was stirred at 80° C. overnight. After concentration in vacuum, the residue was dissolved in DCM and washed with saturated aqueous NaHCO3. The separated organic layer was dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=95:5:0.1) to provide the title compound (0.102 g, 28% over three steps) as a light brown solid. 1 H NMR (400MHz, DMSO-d6): δ8.19(1H,d,J=2.0Hz),8.10(1H,d,J=1.6Hz),7.96(2H,m),7. 77(1H,t,J=6.4Hz),7.61(1H,d,J=2.0Hz),7.35(1H,d,J=1.6Hz),7.30(1H,brs),5.59( 2H,t,J=4.4Hz),4.78(2H,d,J=1.6Hz),4.10(4H,m),3.83(3H,s),3.80(3H,s),3.47-3 .42(2H,m),3.07(2H,t,J=9.6Hz),2.66(1H,m),1.68-1.63(2H,m),1.38-1.37(11H,m).

[0362] Step E: (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0363] NaOH (28.0 mg, 0.704 mmol) was added to a solution of methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate (45.0 mg, 0.0470 mmol) in a mixture of MeOH (0.30 mL), THF (0.30 mL) and water (0.30 mL) at room temperature. The reaction mixture was stirred at 60° C. for 2 hours and cooled to room temperature. After concentration in vacuo, the residue was acidified with citric acid to pH 2. The precipitated solid was collected by filtration, washed with water and dried under vacuum to provide the title compound (38.0 mg, 86%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ7.89(1H,s),7.76(1H,s),7.66(1H,s),7.37(2H,s),7.2 7(1H,s),6.53(1H,s),6.51(1H,s),5.90-5.87(2H,m),4.91(s,4H),4.68(s,2H),4 .53(4H, d, J=6.8Hz), 3.65(3H, s), 3.45-3.42(2H, m), 2.93-2.90(2H, m), 2.15-2. 13(1H, m), 2.10 and 2.08(6H, s+s), 1.54(2H, brs), 1.34(9H, s), 1.29-1.22(8H, m).

[0364]

[0365] Example 2: tert-Butyl (E)-4-(3-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-5-(methoxycarbamoyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0366] [ka]

[0367] (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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-carboxamide) in DMF (2.0 mL) at room temperature To a solution of o-methylhydroxylamine hydrochloride (17.0 mg, 0.201 mmol), EDC (39.0 mg, 0.201 mmol), HOBT (6.16 mg, 0.0400 mmol) and DIPEA (0.0350 mL, 0.201 mmol) were added successively. The reaction mixture was stirred at room temperature overnight. After dilution with a mixture of DCM / MeOH, the mixture was washed with water, followed by brine, then dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:0.1 to 90:10:0.1) followed by recrystallization from hexane / acetone to provide the title compound (10.0 mg, 26%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ11.74(1H,s),7.90(1H,s),7.66(1H,s),7.57(1H,s),7.38(1H ,s),7.35(1H,s),7.11(1H,s),6.53(2H,s),5.94-5.82(2H,m),4.91(4H,s),4.65(2H,s) ,4.53(4H,d,J=6.4Hz),3.73(3H,s),3.63(3H,s),3.50-3.42(2H,m),2.93-2.84(2H,m), 2.44(1H,s),2.11(6H,d,J=3.6Hz),1.60-1.51(2H,m),1.34(11H,s),1.30-1.23(6H,m). LC-MS:m / z=974.3[M+H] + .

[0368]

[0369] Example 3: tert-Butyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0370] [ka]

[0371] (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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 A mixture of 1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid (Example 1, 20.0 mg, 0.0210 mmol), NH4Cl (11.3 mg, 0.212 mmol), HATU (12.1 mg, 0.0320 mmol) and DIPEA (0.0110 mL, 0.0630 mmol) was stirred at 120° C. for 2 h. After concentration in vacuum, the residue was purified by column chromatography on NH-SiO2 (DCM:MeOH=93:7) to provide the title compound (12.4 mg, 62%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.98(1H,s),7.91(1H,s),7.65(2H,d,J=3.6Hz),7.3 6(3H,s),7.28(1H,s),6.54(1H,s),6.52(1H,s),5.94-5.82(2H,m),4.91(4H, s),4.67(2H,s),4.54-4.52(4H,m),3.66(3H,s),3.46-3.42(2H,m),2.44(1H, s),2.10(6H,d,J=4.4Hz),1.57-1.54(2H,m),1.34(9H,s),1.29-1.24(10H,m). LC-MS: m / z=944.3[M+H] + .

[0372]

[0373] Example 4: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride

[0374] [ka]

[0375] HCl (4M HCl in dioxane, 0.622 mL, 2.49 mmol) was added to a solution of tert-butyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate (Example 3, 47.0 mg, 0.0500 mmol) in DCM (2.5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h. After concentration in vacuo, the residual solid was purified by recrystallization from Et2O / MeOH. The solid was collected by filtration, washed with Et2O, and dried under vacuum to provide the title compound (32.0 mg, 73%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ8.69(1H,s),8.58(1H,s),8.02(1H,s),7.96(1H,s),7.67(1H,d,J=0.8Hz),7.65(1H,d ,J=0.8Hz),7.40(1H,d,J=1.6Hz),7.38(2H,s),7.31(1H,d,J=1.2Hz),6.55(1H,s),6.54(1H,s),5.93-5.79(2H ,m),4.91(4H,d,J=4.8Hz),4.73(2H,s),4.53(4H,q,J=7.1Hz),3.69(3H,s),3.05-3.03(2H,m),2.84-2.77(2H, m),2.65-2.58(1H,m),2.11(6H,d,J=1.6Hz),1.83-1.78(2H,m),1.60-1.51(2H,m),1.28(6H,td,J=1.3,7.1Hz). LC-MS:m / z=844.4[M+H] + .

[0376]

[0377] Example 5: Isopropyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0378] [ka]

[0379] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 17.0 mg, 0.0190 mmol) in DCM (0.97 mL) at room temperature was added isopropyl carbonochloridate (9.65 μL, 0.0190 mmol) and DIPEA (10.1 μL, 0.0580 mmol). The reaction mixture was stirred at room temperature for 20 min. After concentration in vacuo, the residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to afford the title compound (7.1 mg, 40%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ12.78(2H,brs),7.97(1H,s),7.90(1H,s),7.65(2H,s),7.36 (3H,s),7.28(1H,s),6.54(1H,s),6.52(1H,s),5.94-5.82(2H,m),4.91(4H,s),4.74-4 .68(3H,m),4.56-4.50(4H,d,m),3.67(3H,s),3.49-3.45(2H,m),2.95(2H,t,J=10.0H z),2.14-2.10(6H,m),1.57-1.54(2H,m),1.27(8H,t,J=7.2Hz),1.13(6H,d,J=6.4Hz). LC-MS:m / z=930.3[M+H] + .

[0380]

[0381] Example 6: Cyclopropyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0382] [ka]

[0383] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) in DMF (0.23 mL) at room temperature was added DIPEA (19.8 μL, 0.114 mmol) and cyclopropyl(4-nitrophenyl)carbonate (Intermediate 7, 5.07 mg, 0.0230 mmol). The reaction mixture was stirred at room temperature for 1 h. After treatment with Et2O, the precipitated solid was collected by filtration and purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=97:7:1) to provide the title compound (15 mg, 72%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6): δ 7.98 (1H,s), 7.90 (1H,s), 7.65 (2H,d,J=2.4 Hz), 7.36 (3H,s), 7.28 (1H,s), 6.54 (1H,s), 6.52 (1H,s), 5.94-5.83 (2H,m), 4.91 (4H,s), 4.68 (2H,s), 4.57-4.50 (4H,m), 3.06 (3H,m), 3.04 (3H,m), 3.02 (3H,m), 3.01 ... .95-3.90(1H,m),3.67(3H,s),3.63-3.59(1H,m),3.15-3.10(1H,m),2.96-2.91(2H,m),2.46-2 .42(1H,m),2.11(3H,s),2.10(3H,s),1.59-1.511(2H,m),1.29-1.26(8H,m),0.60-0.55(4H,m). LC-MS:m / z=928.3[M+H] + .

[0384]

[0385] Example 7: Cyclopentyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0386] [ka]

[0387] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (Example 4, 20.0 mg, 0.0240 mmol) in DMF (0.79 mL) at room temperature was added cyclopentyl(4-nitrophenyl)carbonate (Example 9, 6.55 mg, 0.0260 mmol) and DIPEA (0.0210 mL, 0.118 mmol). The reaction mixture was stirred at room temperature for 30 min. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (2.6 mg, 11%) as a light brown solid. 1H NMR (400 MHz, DMSO-d6): δ 12.83 (2H,s), 7.96 (1H,s), 7.89 (1H,s), 7.65 (2H,d,J=3.2Hz), 7.36 (3H,d,J=8.4Hz), 7.28 (1H,s), 6.53 (2H,d,J=5.2Hz), 5.90-5.87 (2H,m), 4. 91(5H,s),4.68(2H,s),4.53(4H,q,J=6.1Hz),3.69(3H,s),2.97-2.92(2H,m),2.52( 1H,s), 2.10(6H,d,J=3.2Hz),1.74(2H,s),1.55-1.48(8H,m),1.27(8H,t,J=7.2Hz). LC-MS:m / z=956.3[M+H] + .

[0388]

[0389] Example 8: 1-(tert-butoxycarbonyl)piperidin-4-yl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0390] [ka]

[0391] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (0.57 mL) at room temperature To a solution of -7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 50.0 mg, 0.0570 mmol) was added DIPEA (49.6 μL, 0.284 mmol) and tert-butyl 4-(((4-nitrophenoxy)carbonyl)oxy)piperidine-1-carboxylate (Intermediate 8, 20.8 mg, 0.0570 mmol). The reaction mixture was stirred at room temperature for 1 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (32 mg, 53%) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ7.98(1H,s),7.91(1H,s),7.65(2H,d,J=3.2Hz),7.36(3H,s),7.27(1H ,s),6.54(1H,s),6.53(1H,s),5.94-5.76(2H,m),4.91(4H,s),4.67(3H,s),4.53(4H,d,J=6.4Hz ),3.65(3H,s),3.49-3.44(4H,m),3.23-3.12(2H,m),3.04-2.93(2H,m),2.11(3H,s),2.10(3H,s ),1.74-1.70(2H,m),1.60-1.54(2H,m),1.45-1.41(2H,m),1.38(11H,s),1.27(6H,t,J=7.2Hz). LC-MS:m / z=1071.5[M+H] + .

[0392]

[0393] Example 9: Piperidin-4-yl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate hydrochloride

[0394] [ka]

[0395] HCl (4M HCl in dioxane, 233 μL, 0.934 mmol) was added to a solution of 1-(tert-butoxycarbonyl)piperidin-4-yl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate (Example 8, 20.0 mg, 0.019 mmol) in DCM (0.93 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h. After concentration in vacuo, the residual solid was recrystallized from Et2O / MeOH. The solid was collected by filtration, washed with Et2O, and dried under vacuum to provide the title compound (10 mg, 53%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ8.74(2H,s),8.01(1H,s),7.94(1H,s),7.66(2H,s),7.39(3H,s),7.3 0(1H,s),6.55(1H,s),6.53(1H,s),5.94-5.81(2H,m),4.92(4H,d,J=4.0Hz),4.79-4.74(1H,m) ,4.70(2H,s),4.53(4H,q,J=5.9Hz),3.68(3H,s),3.08(6H,d,J=22.9Hz),2.67-2.64(1H,m),2. 11(3H,s),2.10(3H,s),1.97-1.91(2H,m),1.76-1.68(2H,m),1.60-1.55(2H,m),1.34-1.26(9H m). LC-MS:m / z=971.4[M+H] + .

[0396]

[0397] Example 10: 2-Hydroxyethyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0398] [ka]

[0399] Step A: 2-((tert-butyldimethylsilyl)oxy)ethyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0400] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (2.0 mL) at room temperature To a solution of 1H-benzo[d]imidazole-5-carboxamide (Example 4, 50.0 mg, 0.0590 mmol), TEA (0.0410 mL, 0.296 mmol) and 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carbonate (Intermediate 10, 40.0 mg, 0.118 mmol) were added. The reaction mixture was stirred at room temperature for 30 min. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (32 mg, 51%) as a light brown solid. 1H NMR (400MHz, DMSO-d): δ12.84(2H,s),7.98(1H,s),7.91(1H,s),7.65-7.65(2H,m),7.36(3 H,s),7.28(1H,s),6.53(2H,d,J=2.8Hz),5.91-5.87(2H,m),4.91(4H,s),4.67(2H,s),4.53 (4H,q,J=6.5Hz),3.99-3.99(2H,m),3.69(2H,t,J=4.8Hz),3.66(3H,s),2.98-2.98(2H,m), 2.09(6H,d,J=3.2Hz),1.56-1.56(2H,m),1.27(8H,t,J=7.0Hz),0.80(9H,s),-0.02(6H,s). LC-MS:m / z=1046.6[M+H] + .

[0401]

[0402] Step B: 2-Hydroxyethyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0403] HCl (4M in dioxane, 0.0360 mL, 0.143 mmol) was added to a solution of 2-((tert-butyldimethylsilyl)oxy)ethyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate (30.0 mg, 0.0290 mmol) in DCM (0.95 mL) at room temperature. The reaction mixture was stirred at room temperature for 30 min. The precipitated solid was suspended in DCM / hexane and collected by filtration. The solid was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (8.0 mg, 29%) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ8.00(1H,s),7.93(1H,s),7.65(2H,d,J=0.8Hz),7.37(3H, s),7.29(1H,s),6.54(2H,d,J=13.2Hz),5.89-5.84(2H,m),4.92-4.91(4H,m),4.68( 2H,s),4.52(4H,q,J=3.8Hz),3.95(2H,t,J=5.2Hz),3.59(2H,s),2.97-2.97(2H,m) ,2.46-2.46(1H,m),2.10(6H,d,J=5.6Hz),1.58-1.56(2H,m),1.28(8H,t,J=3.6Hz). LC-MS:m / z=932.3[M+H] + .

[0404]

[0405] Example 11: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-pivaloylpiperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0406] [ka]

[0407] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 15.0 mg, 0.017 mmol) in DMF (0.85 mL) at room temperature was added pivaloyl chloride (3.13 μL, 0.0260 mmol) and DIPEA (8.93 μL, 0.0510 mmol). The reaction mixture was stirred at room temperature for 20 min. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to afford the title compound (3.8 mg, 24%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.82(2H,brs),7.97(1H,s),7.90(1H,s),7.65(2H,s),7 .37(1H,s),7.34(2H,s),7.28(1H,s),6.54(1H,s),6.52(1H,s),5.93-5.84(2H,m) ,4.91(4H,s),4.68(2H,s),4.53(4H,d,J=6.8Hz),3.66-3.59(5H,m),3.10-3.04(2 H,m),2.11(3H,s),2.10(3H,s),1.63-1.54(2H,m),1.29-1.23(8H,m),1.10(9H,s). LC-MS:m / z=928.3[M+H] + .

[0408]

[0409] Example 12: tert-Butyl (E)-4-(4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carbonyl)piperidine-1-carboxylate

[0410] [ka]

[0411] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)- in DMF (0.22 mL) To a solution of 7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) was added 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (6.25 mg, 0.0270 mmol), EDC (8.71 mg, 0.0450 mmol) and DIPEA (0.020 mL, 0.114 mmol). The reaction mixture was stirred at room temperature overnight and then heated at 60° C. for 5 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (5.1 mg, 21%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.79(2H,brs),7.93(1H,s),7.86(1H,s),7.62(2H,s),7.35(1H,s),7.31(2 H,s),7.25(1H,s),6.51(1H,s),6.49(1H,s),5.91-5.79(2H,m),4.88(4H,s),4.67(2H,s),4.49(4H,d ,J=8.0Hz),3.89-3.83(2H,m),3.65(3H,s),3.60-3.51(2H,m),3.15-2.93(2H,m),2.73-2.62(3H,m), 2.07(3H,s),2.06(3H,s),1.96(1H,q,J=7.6Hz),1.59-1.44(4H,m),1.34(9H,s),1.31-1.22(10H,m). LC-MS:m / z=1055.4[M+H] + .

[0412]

[0413] Example 13: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0414] [ka]

[0415] To a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (5.25 mg, 0.0340 mmol) in DMF (0.45 mL) was added HATU (13.0 mg, 0.0340 mmol), HOBT (5.22 mg, 0.0340 mmol) and TEA (9.50 μL, 0.0680 mmol). The mixture was stirred at room temperature for 5 min. After adding (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol), the reaction mixture was stirred at room temperature for 40 min. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (9.1 mg, 41%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.85(2H,brs),7.99(1H,s),7.93(1H,s),7.66(1H,s),7.65(1H,s),7.3 9(3H,s),7.28(1H,s),6.54(1H,s),6.52(1H,s),6.09(1H,s),5.90-5.86(2H,m),4.91(4H,s),4.7 0(2H,s),4.52(4H,d,J=5.2Hz),3.99(2H,q,J=7.2Hz),3.66(3H,s),3.24-3.17(2H,m),2.58(1H,s ),2.12(3H,s),2.10(3H,s),2.09(3H,s),1.64-1.59(2H,m),1.41-1.37(2H,m),1.28-1.24(9H,m). LC-MS:m / z=980.4[M+H] + .

[0416]

[0417] Example 14: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(cyclopropylcarbamoyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0418] [ka]

[0419] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) in DMF (0.23 mL) at room temperature was added DIPEA (0.0198 mL, 0.114 mmol) and 4-nitrophenyl cyclopropyl carbamate (Intermediate 5, 5.05 mg, 0.0230 mmol). The reaction mixture was stirred at room temperature for 20 min. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (5.3 mg, 25%) as a yellow solid. 1H NMR(400MHz,DMSO-d6):δ12.80(2H,s),7.99(1H,s),7.91(1H,s),7.65(2H,s),7.36(3H,s),7.2 8(1H,s),6.54(1H,s),6.52(1H,s),6.48(1H,d,J=2.8Hz),5.94-5.84(2H,m),4.91(4H,d,J=4.4H z),4.68(2H,s),4.52(4H,s),3.68(3H,s),2.80(2H,t,J=10.2Hz),2.46-2.43(2H,m),2.11(3H, s),2.09(3H,s),1.53-1.47(2H,m),1.27(8H,t,J=7.2Hz),0.51-0.45(2H,m),0.34-0.30(2H,m). LC-MS:m / z=927.2[M+H] + .

[0420]

[0421] Example 15: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(morpholine-4-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0422] [ka]

[0423] To a solution of morpholine-4-carbonyl chloride (3.18 μL, 0.0270 mmol) and TEA (7.92 μL, 0.0570 mmol) in DMF (39 μL) was added (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol). The reaction mixture was heated to 120° C. for 1.5 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (20 mg, 93%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.97(1H,s),7.90(1H,s),7.65(2H,s),7.38(1H,s),7.35 (2H,s),7.29(1H,s),6.54(1H,s),6.52(1H,s),5.94-5.83(2H,m),4.91(4H,s),4.70(2H,s),4.53 (4H,d,J=6.4Hz),3.68(3H,s),3.50(4H,t,J=4.4Hz),3.20-3.17(2H,m),3.02(4H,t,J=4.2Hz),2. 79(2H,t,J=9.6Hz),2.45(1H,s),2.11(3H,s),2.10(3H,s),1.58-1.55(2H,m),1.33-1.26(8H,m). LC-MS:m / z=957.4[M+H] + .

[0424]

[0425] Example 16: tert-Butyl (E)-4-(4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carbonyl)piperazine-1-carboxylate

[0426] [ka] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (0.45 mL) at room temperature To a solution of 1-(tert-butyl)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 40.0 mg, 0.0450 mmol) was added DIPEA (0.040 mL, 0.23 mmol) and 1-(tert-butyl)4-(4-nitrophenyl)piperazine-1,4-dicarboxylate (Intermediate 6, 16.0 mg, 0.0450 mmol). The reaction mixture was stirred for 24 h and then heated at 120° C. for 2 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (25 mg, 53%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.84(2H,s),7.98(1H,s),7.91(1H,s),7.65(2H,d,J=3.6Hz),7.37( 3H,s),7.28(1H,s),6.54(1H,s),6.52(1H,s),5.94-5.82(2H,m),4.91(4H,s),4.68(2H,s),4.5 3(4H,d,J=6.4Hz),3.66(3H,s),3.26(4H,s),3.20-3.17(2H,m),3.00(4H,s),2.79(2H,t,J=9.8 Hz),2.45(1H,s),2.11(3H,s),2.09(3H,s),1.58-1.55(2H,m),1.39(9H,s),1.32-1.25(8H,m). LC-MS:m / z=1056.7[M+H] + .

[0427] Example 17: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(piperazine-1-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride

[0428] [ka]

[0429] To a solution of tert-butyl (E)-4-(4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carbonyl)piperazine-1-carboxylate (Example 16, 10.0 mg, 9.47 μmol) in DCM (0.47 mL) at room temperature was added HCl (4M in dioxane, 0.118 mL, 0.473 mmol). The reaction mixture was stirred for 1 h. After concentration in vacuo, the residual solid was recrystallized from Et2O and MeOH, washed with Et2O, and dried under vacuum to provide the title compound (6.1 mg, 65%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ8.92(2H,s),8.01(1H,s),7.94(1H,s),7.65(2H,t,J=1.0Hz),7.40(1 H,s),7.38(2H,s),7.30(1H,s),6.55(1H,s),6.53(1H,s),5.94-5.81(2H,m),4.91(4H,d,J=4. 8Hz),4.70(2H,s),4.56-4.50(4H,m),3.69(3H,s),3.22(6H,d,J=5.2Hz),3.04(4H,s),2.86-2 .81(2H,m),2.54-2.53(1H,m),2.11(3H,s),2.10(3H,s),1.58-1.55(2H,m),1.35-1.25(8H,m). LC-MS:m / z=956.4[M+H] + .

[0430]

[0431] Example 18: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(4-hydroxypiperidine-1-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0432] [ka]

[0433] Step A: (E)-7-((3-(1-(4-((tert-butyldimethylsilyl)oxy)piperidine-1-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0434] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (0.73 mL) at room temperature To a solution of -7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 64.0 mg, 0.0730 mmol) was added DIPEA (0.064 mL, 0.36 mmol) and 4-nitrophenyl-4-((tert-butyldimethylsilyl)oxy)piperidine-1-carboxylate (Intermediate 11, 30.4 mg, 0.0800 mmol). The reaction mixture was stirred at 120° C. overnight. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (12 mg, 16%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.84(2H,brs),7.96(1H,s),7.90(1H,s),7.66(2H,d,J=3.6Hz),7.37(1H,s),7.35( 2H,s)7.28(1H,s),6.54(1H,s),6.53(1H,s),5.94-5.81(2H,m),4.92(4H,s),4.69(2H,s),4.53(4H,q,J=6.7Hz ),3.83-3.77(1H,m),3.66(3H,s),3.27-3.23(2H,m),3.19-3.15(2H,m),2.84(2H,t,J=9.2Hz),2.76(2H,t,J=9 .8Hz),2.44(1H,s),2.11(3H,s),2.10(3H,s),1.63-1.56(4H,m),1.32-1.26(10H,m),0.85(9H,s),0.03(6H,s)

[0435] Step B: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(4-hydroxypiperidine-1-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0436] HCl (4M in dioxane, 0.14 mL, 0.56 mmol) was added to a solution of (E)-7-((3-(1-(4-((tert-butyldimethylsilyl)oxy)piperidine-1-carbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide (12.2 mg, 0.0110 mmol) in DCM (0.38 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The precipitated solid was recrystallized from Et2O and MeOH, washed with Et2O, and dried under vacuum to provide the title compound (7.3 mg, 67%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.83(2H,brs),7.97(1H,s),7.90(1H,s),7.66(2H,d,J=2.0Hz),7.37(1 H,s),7.35(2H,s),7.29(1H,s),6.55(1H,s),6.53(1H,s),5.96-5.83(2H,m),4.92(4H,s),4.68(2 H,s),4.53(4H,q,J=3.2Hz),3.67(3H,s),3.23-3.26(2H,m),3.19-3.16(2H,m),2.78-2.72(4H,m) ,2.42(1H,s),2.11(3H,s),2.10(3H,s),1.67-1.63(2H,m),1.58-1.55(2H,m),1.34-1.21(10H,m). LC-MS:m / z=971.2[M+H] + .

[0437]

[0438] Example 19: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(methylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0439] [ka]

[0440] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (7.0 μL, 0.050 mmol) in DCM (0.16 mL) at 0° C. was added MsCl (3.54 μL, 0.0450 mmol). The reaction mixture was stirred for 4 h at temperature. After concentration in vacuo, the residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (2.9 mg, 14%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.97(1H,s),7.92(1H,s),7.65(2H,s),7.37(3H,s),7.28(1 H,s),6.55(1H,s),6.52(1H,s),5.94-5.83(2H,m),4.92(4H,d,J=3.6Hz),4.68(2H,s) ,4.54(4H,t,J=5.6Hz),3.67(3H,s),3.17-3.13(2H,m),2.79-2.76(5H,m),2.43-2.41 (1H,m),2.11(3H,s),2.10(3H,s),1.71-1.67(2H,m),1.46-1.40(2H,m),1.28(6H,m). LC-MS:m / z=922.3[M+H] + .

[0441]

[0442] Example 20: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(isopropylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0443] [ka]

[0444] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.11 mmol) in DMF (0.16 mL) at 0° C. was added propane-2-sulfonyl chloride (5.06 μL, 0.0450 mmol). The reaction mixture was stirred at room temperature for 3 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (1.0 mg, 5%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ 7.97(1H,s), 7.91(1H,s), 7.66(1H,s), 7.65(1H,s), 7.38(1H,s), 7.36(2H,s), 7.29(1H,s), 6.55(1H,s), 6.53(1H,s), 5.95-5.84(2H,m), 4.92(4H,s), 4.69(2H, s),4.54(4H,s),3.67(3H,s),3.21-3.11(3H,m),2.93(2H,t,J=9.0Hz),2.12(3H,s),2.10(3 H,s),1.65-1.60(2H,m),1.39-1.33(2H,m),1.28(6H,t,J=7.2Hz),1.10(3H,s),1.09(3H,s). LC-MS:m / z=950.3[M+H] + .

[0445]

[0446] Example 21: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(cyclopropylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0447] [ka]

[0448] Cyclopropanesulfonyl chloride (4.63 μL, 0.0450 mmol) was added to a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.11 mmol) in DMF (0.16 mL) at 0° C. The reaction mixture was stirred at room temperature for 1 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (1.1 mg, 5%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ 7.97 (1H,s), 7.92 (1H,s), 7.66 (2H,s), 7.38 (1H,s), 7.37 (2H,s), 7.29 (1H,s), 6.55 (1H,s), 6.53 (1H,s), 5.95-5.85 (2H,m), 4.92 (4H,s), 4.68 (2H,s), 4.54 (4H,t,J=5.7Hz), 3.68(3H,s),3.17(2H,s),2.86(2H,t,J=8.9Hz),2.46-2.41(1H,m),2.38-2.32(1H,m),2.12(3H,s) ,2.10(3H,s),1.70-1.64(2H,m),1.46-1.38(2H,m),1.28(6H,td,J=7.1,1.5Hz),0.88-0.84(4H,m). LC-MS:m / z=948.2[M+H] + .

[0449]

[0450] Example 22: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(cyclopentylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0451] [ka]

[0452] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 15.0 mg, 0.0180 mmol) in DMF (1.7 mL) at room temperature was added DIPEA (0.0190 mL, 0.107 mmol) and cyclopentanesulfonyl chloride (9.37 μL, 0.0710 mmol). The reaction mixture was stirred at room temperature for 1 h. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (6.7 mg, 38%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.85(2H,s),7.97(1H,s),7.92(1H,s),7.66-7.65(2H,m),7.39(1H,s ),7.37(2H,s),7.29(1H,s),6.54(2H,d,J=8.0Hz),5.97-5.85(2H,m),4.92(4H,s),4.70(2H,s), 4.58-4.51(4H,m),3.67(3H,s),3.10-3.08(1H,m),2.92-2.87(1H,m),2.10(6H,d,J=7.6Hz),1. 74-1.68(2H,m),1.66-1.52(6H,m),1.50-1.43(2H,m),1.40-1.35(2H,m),1.28(6H,t,J=6.9Hz). LC-MS:m / z=976.3[M+H] + .

[0453]

[0454] Example 23: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-tosylpiperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0455] [ka]

[0456] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.11 mmol) in DMF (0.16 mL) at 0° C. was added p-TsCl (8.66 mg, 0.0450 mmol). The reaction mixture was stirred at room temperature for 20 min. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to afford the title compound (9.6 mg, 42%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ12.85(2H,s),7.97(1H,s),7.90(1H,s),7.66(1H,s),7.65(1H,s),7.53(1H, s),7.51(1H,s),7.38(2H,s),7.36(2H,s),7.30(1H,s),7.26(1H,s),6.57(1H,s),6.51(1H,s),5.86- 5.73(2H,m),4.85(4H,dd,J=18.2,4.2Hz),4.58-4.48(6H,m),3.62(3H,s),3.10-3.01(2H,m),2.37(1 H,s),2.33(3H,s),2.12(3H,s),2.09(3H,s),1.71-1.62(2H,m),1.45-1.38(2H,m),1.31-1.27(6H,m). LC-MS:m / z=998.4[M+H] + .

[0457]

[0458] Example 24: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(N,N-dimethylsulfamoyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0459] [ka]

[0460] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.11 mmol) in DMF (0.16 mL) at 0° C. was added dimethylsulfamoyl chloride (4.88 μL, 0.0450 mmol). The reaction mixture was stirred at room temperature for 1 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (8.4 mg, 39%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ 7.97(1H,s), 7.91(1H,s), 7.66(1H,s), 7.65(1H,s), 7.36(3H,s), 7.28(1H,s), 6.55(1H,s), 6.53(1H,s), 5.91-5.85(2H,m), 4.92(4H,s), 4.67(2H,s), 4.54(4H ,d,J=6.4Hz),3.66(3H,s),3.26-3.18(2H,m),2.86-2.80(2H,m),2.65(6H,s),2.47-2.42(1 H,m),2.12(3H,s),2.10(3H,s),1.68-1.62(2H,m),1.41-1.33(2H,m),1.28(6H,t,J=7.0Hz). LC-MS:m / z=951.2[M+H] + .

[0461]

[0462] Example 25: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((3-(1-(N,N-diethylsulfamoyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0463] [ka]

[0464] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.114 mmol) in DMF (0.16 mL) at 0° C. was added diethylsulfamoyl chloride (5.83 μL, 0.0450 mmol) dropwise. The reaction mixture was stirred at room temperature for 3 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (9.4 mg, 42%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ7.98(1H,s),7.92(1H,s),7.67(1H,s),7.65(1H,s),7.37(2H,s),7.36 (1H,s),7.28(1H,s),6.55(1H,s),6.53(1H,s),5.89-5.85(2H,m),4.91(4H,s),4.66(2H,s),4.5 3(4H,d,J=6.0Hz),3.65(3H,s),3.10-3.04(6H,m),2.72(2H,t,J=9.2Hz),2.46(1H,s),2.11(3H, s),2.10(3H,s),1.66-1.63(2H,m),1.42-1.37(2H,m),1.29-1.26(6H,m),1.01(6H,t,J=7.0Hz). LC-MS:m / z=979.3[M+H] + .

[0465] Example 26: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(pyrrolidin-1-ylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0466] [ka]

[0467] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 20.0 mg, 0.0230 mmol) and TEA (0.016 mL, 0.11 mmol) in DMF (0.16 mL) at 0° C. was added pyrrolidine-1-sulfonyl chloride (5.21 μL, 0.0450 mmol). The reaction mixture was stirred at room temperature for 30 min. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (9.5 mg, 43%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ12.82(2H,s),7.97(1H,s),7.91(1H,s),7.67(1H,s),7.65(1H,s),7.36(3H, s),7.28(1H,s),6.55(1H,s),6.53(1H,s),5.94-5.82(2H,m),4.92(4H,s),4.67(2H,s),4.54(4H,d,J =4.0Hz),3.66(3H,s),3.17-3.14(2H,m),3.07(4H,t,J=6.6Hz),2.83-2.77(2H,m),2.44(1H,s),2.11 (3H,s),2.10(3H,s),1.79-1.73(4H,m),1.66-1.63(2H,m),1.43-1.35(2H,m),1.28(6H,t,J=7.2Hz). LC-MS:m / z=977.5[M+H] + .

[0468]

[0469] Example 27: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(morpholinosulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide

[0470] [ka]

[0471] To a solution of (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide (Example 4, 15.0 mg, 0.0180 mmol) in DMF (1.7 mL) at room temperature was added TEA (0.0150 mL, 0.107 mmol) and morpholine-4-sulfonyl chloride (5.39 μL, 0.0440 mmol). The reaction mixture was stirred at room temperature for 30 min. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (6.2 mg, 35%) as a light brown solid. 1H NMR (400MHz, DMSO-d): δ12.85(2H,s),7.97(1H,s),7.91(1H,s),7.65(2H,s),7.36(3H,s),7.28(1 H,s),6.55(1H,s),6.53(1H,s),5.91-5.87(2H,m),4.92(4H,brs),4.67(2H,s),4.54-4.53(4H,m), 3.67(3H,s),3.55(4H,t,J=4.6Hz),3.24-3.21(2H,m),2.99(4H,t,J=4.6Hz),2.88(2H,t,J=9.0Hz ),2.46(1H,s),2.11(6H,d,J=6.0Hz),1.64-1.60(2H,m),1.39-1.37(2H,m),1.28(6H,t,J=7.2Hz). LC-MS:m / z=993.3[M+H] + .

[0472]

[0473] Example 28: tert-Butyl (E)-4-((4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidin-1-yl)sulfonyl)piperazine-1-carboxylate

[0474] [ka]

[0475] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (0.41 mL) at 0° C. To a solution of tert-butyl 4-(chlorosulfonyl)piperazine-1-carboxylate (Intermediate 4, 32.3 mg, 0.114 mmol) was added tert-butyl 4-(chlorosulfonyl)piperazine-1-carboxylate (Intermediate 4, 32.3 mg, 0.114 mmol) of benzo[d]imidazole-5-carboxamide hydrochloride (Example 4, 50.0 mg, 0.0570 mmol) and TEA (0.040 mL, 0.28 mmol). The reaction mixture was stirred at room temperature for 1 h. After dilution with DCM / MeOH, the mixture was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:1) to provide the title compound (46 mg, 74%) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ12.84(2H,brs),7.98(1H,s),7.91(1H,s),7.66(1H,s),7.65(1H,s),7.37(2H, s),7.34(1H,s),7.27(1H,s),6.56(1H,s),6.53(1H,s),5.93-5.82(2H,m),4.91(4H,d,J=4.0Hz),4.63( 2H,s),4.53(4H,d,J=6.0Hz),3.63(3H,s),3.28-3.23(2H,m),3.02(4H,t,J=5.0Hz),2.83(2H,t,J=9.6H z),2.42(1H,s),2.12(3H,s),2.10(3H,s),1.66-1.64(2H,m),1.41-1.35(11H,m),1.28(6H,t,J=7.0Hz). LC-MS:m / z=1092.4[M+H] + .

[0476]

[0477] Example 29: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((3-(1-(piperazin-1-ylsulfonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride

[0478] [ka]

[0479] To a solution of tert-butyl (E)-4-(4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidin-1-yl)sulfonyl)piperazine-1-carboxylate (Example 28, 20.0 mg, 0.0180 mmol) in DCM (0.92 mL) at room temperature was added HCl (4M in dioxane, 0.23 mL, 0.92 mmol). The reaction mixture was stirred for 1.5 h. After concentration in vacuo, the residual solid was recrystallized from Et2O and MeOH, washed with Et2O, and dried under vacuum to provide the title compound (12 mg, 64%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ9.00(2H,s),8.02(1H,s),7.96(1H,s),7.66(2H,dd,J=3.4,1.4Hz),7.39 (3H,s),7.30(1H,s),6.56(1H,s),6.52(1H,s),5.94-5.82(2H,m),4.92(4H,d,J=4.4Hz),4.70(2H, s),4.54-4.50(4H,m),3.68(3H,s),3.31-3.23(6H,m),3.14(4H,s),2.86(2H,t,J=9.6Hz),2.43(1 H,s),2.11(3H,s),2.10(3H,s),1.67-1.64(2H,m),1.43-1.35(2H,m),1.28(6H,td,J=7.1,1.2Hz). LC-MS:m / z=992.3[M+H] + .

[0480]

[0481] Example 30: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0482] [ka]

[0483] Step A: tert-Butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0484] The title compound was prepared in a similar manner to Example 1 (Step A) using methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (Intermediate 1) and tert-butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate (Intermediate 12). The crude product was purified by column chromatography on SiO2 (DCM:MeOH=99:1-20:1) to provide the title compound (55%) as a red foam. 1H NMR (400MHz, DMSO-d6): δ8.17(1H,d,J=2.0Hz),8.09(1H,d,J=2.0Hz),7.97(1H,d,J=1.6Hz),7.75(1H,t,J=6.4Hz),7.58(1H,d,J=1.6Hz),7.33 (2H,d,J=1.6Hz),5.57(2H,s),4.81(2H,d,J=1.2Hz),4.06-4.04(6H,m) ,3.83(3H,s),3.79(3H,s),3.70-3.66(3H,m),3.58(1H,s),1.34(9H,s). LC-MS:m / z=669.1[M+H] + .

[0485] Step B: tert-Butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0486] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate. The crude product was used in the next reaction without purification as a white foam. LC-MS: m / z=609.02 [M+H] + .

[0487] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0488] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)azetidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=659.2 [M+H] + .

[0489] Step D: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0490] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (37% over three steps) as a light brown oil. 1H NMR (400MHz, DMSO-d6): δ12.86(2H,brs),7.91(1H,s),7.76(1H,d,J=0.8Hz),7.66(1H,s ),7.34(2H,s),7.21(1H,s),6.55(1H,s),6.47(1H,s),5.92-5.83(2H,m),4.91(4H,d,J= 3.6Hz),4.69(2H,s),4.54-4.50(4H,m),3.97(2H,t,J=8.2Hz),3.86(3H,s),3.64-3.63( 1H,m),3.61(2H,s),3.56(3H,s),2.10(6H,d,J=4.4Hz),1.31(9H,s),1.30-1.24(6H,m). LC-MS:m / z=931.3[M+H] + .

[0491] Step E: (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0492] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was suspended in acetone / hexanes, collected by filtration, washed with hexanes, and dried under vacuum to provide the title compound (37%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.98(1H,s),7.73(1H,s),7.66(1H,s ),7.36(2H,s),7.27(1H,d,J=1.2Hz),6.54(1H,s),6.51(1H,s),5.87-5.83( 2H,m),4.91(4H,d,J=4.0Hz),4.70(2H,s),4.52(4H,t,J=6.6Hz),3.98(2H,t ,J=8.6Hz),3.67-3.61(5H,m),2.11(6H,s),1.31(9H,s),1.29-1.25(6H,m). LC-MS: m / z=917.3[M+H] + .

[0493]

[0494] Step F: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)azetidine-1-carboxylate

[0495] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)azetidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (23%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ12.83(2H,d,J=7.3Hz),7.96(1H,s),7.92(1H,s),7.66(1H ,s),7.64(1H,d,J=0.9Hz),7.39(1H,s),7.35(2H,s),7.28(1H,s),6.53(2H,s),5.9 3-5.82(2H,m),4.92(4H,s),4.76(2H,s),4.53(4H,q,J=6.3Hz),3.98(2H,t,J=8.2H z),3.68(3H,s),3.62(2H,t,J=7.0Hz),2.10(6H,s),1.31(9H,s),1.29-1.25(6H,m). LC-MS:m / z=916.3[M+H] + .

[0496]

[0497] Example 31: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0498] [ka]

[0499] Step A: tert-Butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0500] The title compound was prepared in a similar manner to Example 1 (Step A) using tert-butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate (Intermediate 13) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1). The crude product was purified by column chromatography on NH-SiO2 (DCM only to DCM:MeOH=98:2) to provide the title compound (77%) as an orange solid. LC-MS: m / z=683.2 [M+H] + .

[0501] Step B: tert-Butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0502] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=623.2 [M+H] + .

[0503] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0504] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was recrystallized from Et2O / MeOH, washed with Et2O and dried under vacuum to provide the title compound (82%) as a yellow solid. LC-MS: m / z=673.2 [M+H] + .

[0505] Step D: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0506] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=95:5:1) to provide the title compound (31%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.86(2H,s),7.87(1H,s),7.77(1H,d,J=1.2Hz) ,7.67(1H,s),7.35(1H,s),7.31(1H,s),7.20(1H,s),6.55(1H,s),6.50(1 H,s),5.91-5.80(2H,m),4.93-4.89(4H,m),4.62(2H,s),4.56-4.51(4H,m ),3.86(3H,s),3.59(3H,s),3.09-2.92(3H,m),2.11(6H,s),1.99-1.91(1H ,m),1.70-1.61(1H,m),1.34(10H,d,J=4.8Hz),1.28(6H,dd,J=12.2,7.0Hz).

[0507] Step E: (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0508] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was collected by filtration, washed with water, and dried under vacuum to provide the title compound (91%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.85(2H,d,J=11.6Hz),7.89(1H,s),7.76(1H,d,J=1.2Hz),7.66( 1H,s),7.36(1H,s),7.34(1H,s),7.27(1H,s),6.53(1H,s),6.50(1H,s),5.93-5.82(2H,m), 4.92(4H,s),4.70(2H,s),4.53(4H,q,J=6.8Hz),3.66(3H,s),3.11-2.92(3H,m),2.10(6H,s ),1.99-1.92(1H,m),1.70-1.62(1H,m),1.33(10H,d,J=6.0Hz),1.27(6H,td,J=7.1,3.5Hz).

[0509] Step F: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)pyrrolidine-1-carboxylate

[0510] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=93:7) to provide the title compound (63%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.95(1H,s),7.90(1H,s),7.66(1H,s),7.65(1H,s),7.35(3H ,s),7.27(1H,s),6.53(2H,s),5.94-5.80(2H,m),4.91(4H,s),4.69(2H,s),4.52(4H,q,J=6.4Hz),3 .66(3H,s),3.24-3.15(1H,m),3.11-3.08(1H,m),3.05-3.01(1H,m),2.95-2.90(1H,m),2.10(6H,d, J=2.0Hz), 1.98-1.89(1H,m),1.71-1.64(1H,m),1.34(10H,d,J=6.0Hz),1.27(6H,td,J=7.1,1.9Hz). LC-MS:m / z=930.3[M+H] + .

[0511]

[0512] Example 32: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0513] [ka]

[0514] Step A: tert-Butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0515] The title compound was prepared in a similar manner to Example 1 (Step A) using methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (Intermediate 1) and tert-butyl 3-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (Intermediate 14). The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1) to provide the title compound (>99%) as a red foam. LC-MS: m / z=697.2 [M+H] + .

[0516] Step B: tert-Butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0517] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(3-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=637.3 [M+H] + .

[0518] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0519] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-3-(3-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=687.3 [M+H] + .

[0520] Step D: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0521] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound as a light brown oil. LC-MS: m / z=959.4 [M+H] + .

[0522] Step E: (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0523] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was suspended in acetone / hexane, washed with hexane and dried under vacuum to provide the title compound as a light brown solid. LC-MS: m / z=945.3 [M+H] + .

[0524] Step F: tert-Butyl (E)-3-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0525] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (0.1% over six steps) as a white solid. 1 HNMR (400MHz, DMSO-d6): δ12.85(2H,s),7.97(1H,s),7.92(1H,s),7.65(2H,d,J=6.0Hz),7.35(3H,d ,J=7.6Hz),7.27(1H,s),6.53(2H,d,J=8.0Hz),5.91-5.82(2H,m),4.92(4H,s),4.65(2H,s),4.53(4H ,d,J=6.4Hz),3.64(3H,s),3.49-3.40(1H,m),2.89(2H,t,J=10.2Hz),2.34-2.33(1H,m),2.11-2.10 (6H,m),1.70-1.67(1H,m),1.44-1.43(1H,m),1.33(9H,s),1.27(7H,t,J=6.8Hz),1.19-1.17(1H,m). LC-MS:m / z=944.3[M+H] + .

[0526] Example 33: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0527] [ka]

[0528] Step A: Methyl (E)-4-((4-((4-carbamoyl-2-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate

[0529] The title compound was prepared in a similar manner to Example 1 (Step A) using 4-chloro-3-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-5-nitrobenzamide (Intermediate 16) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1). The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=97:3) to afford the title compound (36%) as an orange solid. 1 H NMR (400MHz, DMSO-d6): δ8.19(1H,d,J=2.0Hz), 8.10(1H,d,J=1.6Hz),7.99-7.94(2H,m),7.77(1H,t,J=6.0Hz),7.59(1H,d,J=1.2Hz),7.3 4(1H,d,J=1.6Hz),7.32(1H,s),5.60(2H,q,J=4.0Hz),5.36(1H,s),4.78(2H,s),4.11-4.07(4H,m),3.83(3H,s),3.80(3H,s),1.32(6H,s).

[0530] Step B: Methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate

[0531] The title compound was prepared in a similar manner to Example 1 (Step B) using methyl (E)-4-((4-((4-carbamoyl-2-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate. The crude product was used in the next reaction without purification as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ7.53(1H,brs),7.01(1H,d,J=1.6Hz),6.97(1H,brs),6.86(1H,d,J=2.0Hz),6.85(1H,d,J=2.0Hz),6.82(1H,d,J=1.6Hz ),5.68-5.66(2H,m),5.34(1H,s),4.79(2H,s),4.71-4.70(4H,m),3.76 (3H,s),3.74(3H,s),3.62-3.58(3H,m),3.62-3.53(3H,m),1.34(6H,s).

[0532] Step C: Methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate 2HBr

[0533] The title compound was prepared in a similar manner to Example 1 (Step C) using methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate. The crude solid product was recrystallized from MeOH / EtO, washed with EtO and dried under vacuum to provide the title compound (82%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ8.77-8.65(4H,m),8.03-7.95(1H,m),7.57(1H,d,J=0.8Hz),7.51(1H,s),7.50(1H,s),7.4 8(1H,s),7.32(1H,s),5.84-5.78(2H,m),5.39(1H,brs),4.86-4.83(6H,m),3.87(3H,s),3.74(3H,s),1.31(6H,s).

[0534] Step D: Methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-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-carboxylate

[0535] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate 2HBr and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The residue was purified by column chromatography on SiO2 (DCM:MeOH=95:5 to 90:10) to provide the title compound (66%) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ7.89(1H,brs),7.75(1H,s),7.66(1H,s),7.40(1H,s),7.36(1H,brs),7.24(1H,s),6.54(1H,s),6.44(1H,s),5.89-5.83( 2H,m),5.33(1H,s),4.92(4H,d,J=5.2Hz),4.77(2H,s),4.55-4.46(4H,m) ,3.86(3H,s),3.65(3H,s),2.11(3H,s),2.08(3H,s),1.27-1.24(12H,m).

[0536] Step E: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-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-carboxylic acid

[0537] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-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-carboxylate. The crude solid product was washed with water and dried under vacuum to provide the title compound (>99%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ7.90(1H,brs),7.74(1H,s),7.66(1H,s),7.42(1H,s),7.35(1H,brs),7.29(1H,s),6.52(1H,s),6.44(1H,s),5.94- 5.81(2H,m),5.34(1H,s),4.93(4H,d,J=3.6Hz),4.54-4.48(4H,m),3. 70(3H,s),3.50(3H,s),2.10(3H,s),2.08(3H,s),1.27-1.22(12H,m).

[0538] Step F: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0539] The title compound was prepared in a similar manner to Example 2 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-hydroxy-4-methylpent-2-yn-1-yl)oxy)-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-carboxylic acid. The crude product was purified by Prep-HPLC to provide the title compound (18%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ11.72(1H,s),7.91(1H,brs),7.66(1H,s),7.55(1H ,s),7.41(1H,s),7.37(1H,brs),7.13(1H,s),6.52(1H,s),6.49(1H,s),5.90 -5.82(2H,m),5.32(1H,brs),4.93-4.89(4H,m),4.80(2H,s),4.52-4.47(4H ,m),3.72(3H,s),3.69(3H,s),2.10(3H,s),2.09(3H,s),1.27-1.25(12H,m). LC-MS: m / z=849.3[M+H] + .

[0540]

[0541] Example 34: 2-(Dimethylamino)ethyl (E)-4-(3-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0542] [ka]

[0543] (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((3-(piperidin-4-yl)prop-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) in DMF (0.28 mL) at room temperature To a solution of 2-(dimethylamino)ethyl(4-nitrophenyl)carbonate hydrochloride (Intermediate 17, 9.08 mg, 0.0310 mmol) was added DIPEA (24.8 μL, 0.142 mmol) followed by 2-(dimethylamino)ethyl(4-nitrophenyl)carbonate hydrochloride (Intermediate 17, 9.08 mg, 0.0310 mmol). The reaction mixture was stirred at room temperature for 2 h. After treatment with saturated aqueous NaHCO3, the precipitated solid was collected by filtration. The solid was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=97:7:1) to provide the title compound (8.3 mg, 31%) as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.78(2H,brs),7.96(1H,s),7.89(1H,s),7.66(2H,dd,J=4.2,1.0Hz),7.37 (1H,s),7.35(2H,s),7.29(1H,s),6.55(1H,s),6.53(1H,s),5.95-5.81(2H,m),4.91(4H,s),4.69(2H, s),4.53(4H,q,J=6.8Hz),4.01(2H,t,J=5.8Hz),3.67(3H,s),3.51-3.46(2H,m),2.98(2H,t,J=9.8Hz ),2.41(2H,t,J=5.8Hz),2.12(6H,s),2.11(3H,s),2.10(3H,s),1.58-1.56(2H,m),1.32-1.24(8H,m). LC-MS:m / z=959.2[M+H] + .

[0544]

[0545] [ka]

[0546] Example 35: Methyl (E)-3-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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)-3H-imidazo[4,5-b]pyridine-6-carboxylate

[0547] [ka]

[0548] Step A: tert-Butyl (E)-4-(3-(5-carbamoyl-2-((4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0549] To a solution of tert-butyl 4-(3-(5-carbamoyl-2-chloro-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (Intermediate 15, 1.00 g, 2.28 mmol) and (E)-2-(4-aminobut-2-en-1-yl)isoindoline-1,3-dione (Intermediate 2, 0.741 g, 3.43 mmol) in n-BuOH (11 mL) at room temperature was added DIPEA (1.99 mL, 11.4 mmol). The reaction mixture was stirred at 140° C. overnight and then concentrated in vacuo to provide the title compound (0.320 g, 22%). 1H NMR(400MHz,DMSO-d6):δ8.19(1H,d,J=1.6Hz),7.95(1H,brs),7.88-7.82(4H ,m),7.78(1H,t,J=6.2Hz),7.65(1H,d,J=2.0Hz),7.30(1H,brs),5.72-5.61( 2H,m),4.84(2H,d,J=1.2Hz),4.13-4.10(4H,m),3.46-3.39(2H,m),3.05(2H, t,J=9.4Hz),2.62(1H,s),1.65-1.61(2H,m),1.37(9H,s),1.34-1.28(2H,m).

[0550] Step B: tert-Butyl (E)-4-(3-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0551] To a solution of tert-butyl (E)-4-(3-(5-carbamoyl-2-((4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (0.320 g, 0.518 mmol) in THF (2.9 mL) at room temperature was added hydrazine hydrate (1.38 mL, 5.70 mmol). The reaction mixture was stirred at 60° C. for 1 h. After concentration in vacuo, the residue was treated with 2N aqueous NaOH solution and then extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (0.269 g, quant.) as a red foam. LC-MS: m / z=488.2 [M+H] + .

[0552] Step C: tert-Butyl (E)-4-(3-(5-carbamoyl-2-((4-((5-carbamoyl-3-nitropyridin-2-yl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0553] To a solution of tert-butyl (E)-4-(3-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (269 mg, 0.552 mmol) in DMF (2.8 mL) was added DIPEA (0.549 mL, 3.14 mmol) and 6-chloro-5-nitronicotinamide (117 mg, 0.579 mmol). The reaction was stirred at room temperature for 1 h. After treatment with saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on SiO2 (DCM only to DCM:MeOH=95:5) to provide the title compound (259 mg, 72%) as an orange solid. 1H NMR(400MHz,DMSO-d6):δ8.89-8.85(2H,m),8.83(1H,t,J=5.8Hz),8.21(1H,d,J=1.2Hz),8.09(1H,s),7. 96(1H,s),7.78(1H,t,J=6.2Hz),7.67(1H,d,J=2.0Hz),7.45(1H,s),7.30(1H,s),5.78-5.68(2H,m),4.8 6(2H,s),4.19(2H,t,J=5.0Hz),4.14(2H,t,J=5.4Hz),3.48-3.39(2H,m),3 .09-3.05(2H,m),2.66-2.63(1H,m),1.67-1.62(2H,m),1.39-1.30(11H,m).

[0554] Step D: tert-Butyl (E)-4-(3-(3-amino-2-((4-((3-amino-5-carbamoylpyridin-2-yl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate

[0555] To a solution of tert-butyl (E)-4-(3-(5-carbamoyl-2-((4-((5-carbamoyl-3-nitropyridin-2-yl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (250 mg, 0.383 mmol) in MeOH (9.1 mL) was added NH4OH (2.98 mL, 19.2 mmol) followed by a solution of sodium hyposulfite (800 mg, 4.60 mmol) in water (3.7 mL). The reaction mixture was stirred at room temperature for 1 h. After concentration in vacuo, the residue was partitioned between water and EtOAc. The separated aqueous layer was extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (270 mg, quant.). LC-MS: m / z=593.2[M+H] + .

[0556] Step E: Methyl (E)-3-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-4-yl)prop-2-yn-1-yl)oxy)-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)-3H-imidazo[4,5-b]pyridine-6-carboxylate

[0557] To a solution of tert-butyl (E)-4-(3-(3-amino-2-((4-((3-amino-5-carbamoylpyridin-2-yl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)prop-1-yn-1-yl)piperidine-1-carboxylate (248 mg, 0.418 mmol) in DMF (14 mL) at 0° C. was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (Intermediate 3, 1.67 mL, 0.837 mmol). The mixture was stirred at room temperature for 1 h. DIPEA (0.438 mL, 2.51 mmol) and EDC (321 mg, 1.67 mmol) were added in one portion, after which the reaction mixture was stirred at 70° C. for 1 h and cooled to room temperature. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residual solid was suspended in EtOAc and filtered off. The filtrate was solidified from Et2O / hexane. The solid was collected by filtration, washed with hexane, and dried under vacuum to provide the title compound (34.0 mg, 8.8%) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.80(2H,brs),8.68(1H,s),8.12(2H,s),7.88(1H,s),7.65( 1H,s),7.52(1H,s),7.40(1H,s),7.33(1H,s),6.52(1H,s),6.50(1H,s),5.93(2H,s),4. 92(2H,s),4.78(4H,s),4.54-4.47(4H,m),3.45-3.39(2H,m),2.96(2H,t,J=10.8Hz),2. 46(1H,s),2.10(3H,s),2.09(3H,s),1.58-1.55(2H,m),1.34(9H,s),1.31-1.25(8H,m). LC-MS:m / z=915.3[M+H] + .

[0558]

[0559] Example 36: tert-Butyl (E)-4-(4-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-5-(methoxycarbamoyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0560] [ka]

[0561] Step A: tert-Butyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0562] The title compound was prepared in a similar manner to Example 1 (Step A) using methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1, 0.989 g, 2.98 mmol) and tert-butyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate (Intermediate 18). The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1) to provide the title compound (46%) as a red foam. 1 H NMR(400MHz,DMSO-d6):δ8.18(1H,d,J=1.8Hz),8.09(1H,d,J=1.8Hz),7.96(1H,t,J=6.4Hz),7.76(1H,t,J=6.2Hz),7.60(1H,d,J= 1.8Hz),7.34(1H,d,J=1.8Hz),7.31(1H,s),5.60-5.57(2H,m),4.84(2H,s),4.09-4.08(4H,m),3.84(3H,s),3.81(3H,s),3.30(3Hm) s), 3.25 (4H, s), 2.31 (4H, s), 1.37 (9H, s). LC-MS: m / z=712.2[M+H] + .

[0563] Step B: tert-Butyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0564] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=652.2 [M+H]+ .

[0565] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0566] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=740.3 [M+H] + .

[0567] Step D: Methyl (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0568] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (27% over four steps) as a brown oil. LC-MS: m / z=974.3 [M+H] + .

[0569] Step E: (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0570] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was suspended in acetone, collected by filtration, washed with acetone and dried under vacuum to provide the title compound as a white solid. LC-MS: m / z=960.4 [M+H] + .

[0571] Step F: tert-Butyl (E)-4-(4-((5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(4-(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-5-(methoxycarbamoyl)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0572] The title compound was prepared in a similar manner to Example 2 using (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (14% between two steps) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ12.83(2H,s),11.73(1H,s),7.90(1H,s),7.67(1H,s),7.56( 1H,s),7.37(2H,s),7.13(1H,s),6.53(2H,s),5.89-5.82(2H,m),4.90(4H,d,J=3.2Hz ),4.75(2H,s),4.53(4H,q,J=6.8Hz),3.73(3H,s),3.68(3H,s),3.20(4H,t,J=4.0Hz) ,3.17(2H,s),2.23(4H,s),2.10(6H,d,J=2.0Hz),1.33(9H,s),1.28(6H,t,J=6.8Hz). LC-MS:m / z=989.4[M+H] + .

[0573]

[0574] Example 37: tert-Butyl 4-(4-(2-chloro-5-(methoxycarbonyl)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0575] [ka]

[0576] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid (Example 36, Step E). The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (8%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.97(1H,s),7.92(1H,s),7.67(1H,s),7.65(1H,s),7.3 8(1H,s),7.35(2H,s),7.29(1H,s),6.52(2H,d,J=3.2Hz),5.89-5.79(2H,m),4.91 (4H,d,J=3.2Hz),4.76(2H,s),4.52(4H,q,J=6.4Hz),3.70(3H,s),3.19-3.17(6H ,m),2.23-2.21(4H,m),2.09(6H,d,J=2.4Hz),1.33(9H,s),1.27(6H,t,J=6.8Hz). LC-MS: m / z=959.3[M+H] + .

[0577]

[0578] Example 38: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0579] [ka]

[0580] Step A: Isopropyl (E)-4-(4-(2-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0581] To a solution of isopropyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate (Intermediate 19, 600 mg, 1.39 mmol) in DMSO (6.0 mL) at room temperature was added DIPEA (1.40 mL, 8.04 mmol) and tert-butyl (E)-(4-aminobut-2-en-1-yl)carbamate (520 mg, 2.79 mmol). The reaction mixture was stirred in a sealed tube at 120 °C for 3 h. After concentration in vacuum, the residue was dissolved in DCM and washed with saturated aqueous NaHCO3. The separated organic layer was washed with brine, dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography on NH-SiO2 (DCM:MeOH=20:1) to provide the title compound as a brown oil. LC-MS: m / z=589.2[M+H] + .

[0582] Step B: Isopropyl (E)-4-(4-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0583] To a solution of isopropyl (E)-4-(4-(2-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate (800 mg, 1.36 mmol) in MeOH (11 mL) at 0° C. was added HCl (4 M in dioxane, 1.70 mL, 6.80 mmol). The reaction mixture was stirred at room temperature for 3 h. After concentration in vacuo, the residue was diluted with EtOAc and basified with 2N aqueous NaOH to pH 9. The mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (582 mg, crude). LC-MS: m / z=489.2 [M+H] + .

[0584] Step C: Isopropyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0585] A mixture of isopropyl (E)-4-(4-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate (582 mg, 1.19 mmol), methyl 4-chloro-3-methoxy-5-nitrobenzoate (585 mg, 2.38 mmol) and DIPEA (1.00 mL, 5.74 mmol) in DMF (5.80 mL) was stirred at 100 °C for 3 h in a sealed tube. After quenching the reaction using saturated aqueous NaHCO3, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on NH-SiO2 (DCM:MeOH=50:1) to afford the title compound (513 mg, 52% over three steps) as an orange solid. 1 H NMR(400MHz,CDCl3):δ8.42(1H,s),8.13(1H,s),7.59(1H,s),7.43(1H,s),5.74(2H,brs),4.89(1H,t,J=6.0Hz), 4.78(2H,s),4.26(4H,s),3.91(3H,s),3.85(3H,s),3.44(4H,s),3.34(2H,s),2.40(4H,s),1.23(6H,d,J=6.0Hz).

[0586] Step D: Isopropyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperazine-1-carboxylate

[0587] The title compound was prepared in a similar manner to Example 1 (Step B) using isopropyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitro-phenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude product was used in the next reaction without purification. LC-MS: m / z=638.2 [M+H]+ .

[0588] Step E: Methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0589] The title compound was prepared in a similar manner to Example 1 (Step C) using isopropyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxy-carbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperazine-1-carboxylate. The crude solid product was recrystallized from Et2O / MeOH to provide the title compound as a yellow solid. LC-MS: m / z=688.2 [M+H] + .

[0590] Step F: Methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-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-carboxylate

[0591] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]-imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:TEA=100:10:1) to provide the title compound as a pale yellow solid. LC-MS: m / z=960.1 [M+H] + .

[0592] Step G: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0593] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-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-carboxylate. The crude product was used in the next reaction without purification. LC-MS: m / z=946.1 [M+H] + .

[0594] Step H: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0595] The title compound was prepared in a similar manner to Example 2 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-(isopropoxycarbonyl)piperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]-imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by preparative TLC (NH-SiO2) (DCM:MeOH=10:1) to provide the title compound (6% over five steps) as a yellow solid. 1 H NMR(400MHz,DMSO-d6):δ7.88(1H,s),7.64(1H,s),7.54(1H,s),7.34(2H,brs) ,7.10(1H,s),6.49(2H,s),5.90-5.79(2H,m),4.88-4.87(5H,m),4.74-4.72(2H ,m),4.55-4.49(4H,m),3.72(3H,s),3.66(3H,s),3.57-3.56(2H,m),3.17(2H, brs),2.25-2.23(4H,m),2.09(6H,s),1.26-1.25(6H,m),1.11(6H,d,J=6.4Hz). LC-MS: m / z=975.2[M+H] + .

[0596]

[0597] Example 39: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0598] [ka]

[0599] Step A: Methyl (E)-4-((4-((4-carbamoyl-2-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (9) (K00803-022)

[0600] The title compound was prepared in a similar manner to Example 1 (Step A) using methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1) and 4-chloro-3-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-5-nitrobenzamide (Intermediate 20). The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1) to provide the title compound (36%) as a red foam. 1H NMR(400MHz,DMSO-d6):δ8.18(1H,d,J=1.6Hz),8.10(1H,d,J=2.0Hz),7.97(1H,t,J =6.8Hz),7.76(1H,t,J=6.0Hz),7.60(1H,d,J=2.0Hz),7.35(1H,d,J=1.6Hz),5.59( 2H,dd,J=5.6,4.4Hz),4.85(2H,s),4.09-4.08(4H,m),3.83(3H,s),3.81(3H,s),3. 41(6H,s),2.84-2.76(1H,m),2.37(2H,s),2.32-2.31(2H,m),0.95(6H,d,J=6.4Hz). LC-MS:m / z=682.2[M+H] + .

[0601] Step B: Methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate

[0602] The title compound was prepared in a similar manner to Example 1 (Step B) using methyl (E)-4-((4-((4-carbamoyl-2-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=622.3 [M+H] + .

[0603] Step C: Methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0604] The title compound was prepared in a similar manner to Example 1 (Step C) using methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=672.2 [M+H] + .

[0605] Step D: Methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-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-carboxylate

[0606] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound as a brown oil. LC-MS: m / z=944.2 [M+H] + .

[0607] Step E: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0608] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-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-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=930.4 [M+H] + .

[0609] Step F: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0610] The title compound was prepared in a similar manner to Example 2 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(4-isobutyrylpiperazin-1-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1 to 80:10:1) to provide the title compound (4% over six steps) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.90(1H,s),7.67(1H,s),7.57(1H,s),7.38(2H,s),7 .15(1H,s),6.54(2H,s),5.90-5.83(2H,m),4.89(4H,s),4.75(2H,s),4.54(4H ,q,J=6.8Hz),3.73(3H,s),3.69(3H,s),3.19(2H,s),2.66(1H,q,J=6.4Hz),2. 23(4H,s),2.10(6H,d,J=2.4Hz),1.28(6H,t,J=6.8Hz),0.89(6H,d,J=6.8Hz). LC-MS: m / z=959.3[M+H] + .

[0611]

[0612] Example 40: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-morpholinobut-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0613] [ka]

[0614] Step A: Methyl (E)-4-((4-((4-carbamoyl-2-((4-morpholinobut-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate

[0615] The title compound was prepared in a similar manner to Example 1 (Step A) using methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1) and 4-chloro-3-((4-morpholinobut-2-yn-1-yl)oxy)-5-nitrobenzamide (Intermediate 21). The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=97:3) followed by SiO2 (DCM:MeOH:NH4OH=95:5:0.1) to provide the title compound (30%) as an orange solid. 1 H NMR(400MHz,DMSO-d6):δ8.15(1H,d,J=2.0Hz),8.06(1H,d,J=2.0Hz),7.95-7.90(2H,m),7.72(1H,t,J=6.0Hz),7.58(1H,s),7.31(1H ,s),7.29(1H,brs),5.56(2H,s),4.82(2H,s),4.06(4H,s),3.79(3H,s),3.77(3H,s),3.47(4H,d,J=4.4Hz),3.23(2H,s),2.30(4H,s).

[0616] Step B: Methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-morpholinobut-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate

[0617] The title compound was prepared in a similar manner to Example 1 (Step B) using methyl (E)-4-((4-((4-carbamoyl-2-((4-morpholinobut-2-yn-1-yl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=553.2 [M+H] + .

[0618] Step C: Methyl (E)-2-amino-1-(4-(2-amino-5-carbamoyl-7-((4-morpholinobut-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide

[0619] The title compound was prepared in a similar manner to Example 1 (Step C) using methyl (E)-3-amino-4-((4-((2-amino-4-carbamoyl-6-((4-morpholinobut-2-yn-1-yl)oxy)phenyl)amino)but-2-en-1-yl)amino)-5-methoxybenzoate. The crude product was purified by recrystallization from Et2O / MeOH, collected by filtration, washed with Et2O, and dried under vacuum to provide the title compound (95%) as a light brown solid. LC-MS: m / z=603.1 [M+H] + .

[0620] Step D: Methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-morpholinobut-2-yn-1-yl)oxy)-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-carboxylate

[0621] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-((l2-bromanyl)-l4-azanyl)-1-(4-(2-((l2-bromanyl)-l4-azanyl)-5-carbamoyl-7-((4-morpholinobut-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=95:5:0.1) to provide the title compound (11%) as a brown solid. LC-MS: m / z=875.2 [M+H] + .

[0622] Step E: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-morpholinobut-2-yn-1-yl)oxy)-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-carboxylic acid

[0623] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-morpholinobut-2-yn-1-yl)oxy)-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-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=861.3 [M+H] + .

[0624] Step F: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-morpholinobut-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-N,7-dimethoxy-1H-benzo[d]imidazole-5-carboxamide

[0625] The title compound was prepared in a similar manner to Example 2 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-morpholinobut-2-yn-1-yl)oxy)-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-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=93:7:0.1 to 90:10:0.1) followed by recrystallization from hexane / acetone to provide the title compound (4%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ12.79(2H,brs),11.73(1H,brs),7.90(1H,s),7.66(1H ,s),7.56(1H,s),7.40(1H,s),7.37(1H,s),7.13(1H,s),6.53(2H,s),5.88-5.84 (2H,m),4.91(4H,s),4.78(2H,s),4.52(4H,q,J=6.8Hz),3.73(3H,s),3.68(3H, s),3.48-3.45(4H,m),3.13(2H,s),2.27(4H,s),2.10(6H,s),1.29-1.23(6H,m). LC-MS: m / z=890.3[M+H] + .

[0626]

[0627] Example 41: tert-Butyl (E)-4-(5-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0628] [ka]

[0629] Step A: tert-Butyl (E)-4-(5-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0630] The title compound was prepared in a similar manner to Example 1 (Step A) using tert-butyl 4-(5-(5-carbamoyl-2-chloro-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate (Intermediate 1) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 22). The crude product was purified by column chromatography on NH-SiO2 (DCM only to DCM:MeOH=98:2) to afford the title compound (>99%) as an orange solid. LC-MS: m / z=740.3 [M+H] + .

[0631] Step B: tert-Butyl (E)-4-(5-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0632] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-4-(5-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=680.3 [M+H] + .

[0633] Step C: Methyl (E)-2-amino-1-(4-(2-amino-7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0634] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-4-(5-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=730.3 [M+H] + .

[0635] Step D: Methyl (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0636] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-((l2-bromanyl)-l4-azanyl)-1-(4-(2-((l2-bromanyl)-l4-azanyl)-7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=95:5:0.1) to provide the title compound (7% over four steps) as a brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.85(2H,s),7.88(1H,s),7.76(1H,d,J=1.2Hz),7.67(1H,s) ,7.36(2H,s),7.23(1H,d,J=1.6Hz),6.56(1H,s),6.47(1H,s),5.88-5.79(2H,m),4.90( 4H,d,J=4.8Hz),4.74(2H,s),4.56-4.49(4H,m),3.86(3H,s),3.65(3H,s),3.17(4H,s), 2.31(4H,s),2.11(3H,s),2.09(3H,s),1.32(9H,s),1.28(6H,q,J=7.1Hz),1.19(6H,s).

[0637] Step E: (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0638] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.87(1H,s),12.85(1H,s),7.90(1H,s),7.76(1H,d,J=0.8Hz) ,7.67(1H,s),7.40(1H,s),7.36(1H,s),7.29(1H,d,J=0.8Hz),6.54(1H,s),6.47(1H,s) ,5.94-5.80(2H,m),4.91(4H,s),4.79(2H,s),4.58-4.49(4H,m),3.70(3H,s),3.16(4H, s),2.31(4H,s),2.10(3H,s),2.08(3H,s),1.32(9H,s),1.30-1.25(6H,m),1.18(6H,s).

[0639] Step F: tert-Butyl (E)-4-(5-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)-2-methylpent-3-yn-2-yl)piperazine-1-carboxylate

[0640] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylpent-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=93:7) to provide the title compound (61% between two steps) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.97(1H,s),7.91(1H,s),7.67(1H,s),7.65(1H, s),7.40(1H,s),7.36(2H,s),7.29(1H,d,J=0.8Hz),6.53(1H,s),6.52(1H,s),5.97-5.81 (2H,m),4.91(4H,s),4.78(2H,s),4.53(4H,d,J=6.8Hz),3.70(3H,s),3.17(4H,s),2.31 (4H,s),2.10(3H,s),2.09(3H,s),1.32(9H,s),1.27(6H,td,J=6.1,4.5Hz),1.18(6H,s). LC-MS:m / z=987.5[M+H] + .

[0641]

[0642] Example 42: tert-Butyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0643] [ka]

[0644] Step A: tert-Butyl (E)-4-(4-(5-carbamoyl-2-((4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0645] A mixture of isopropyl 4-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate (Intermediate 23, 650 mg, 1.43 mmol), tert-butyl (E)-(4-aminobut-2-en-1-yl)carbamate (Intermediate 2, 467 mg, 2.15 mmol) and DIPEA (1.30 mL, 7.46 mmol) in n-BuOH (6.5 mL) was irradiated in a microwave at 120° C. for 3 h. The reaction mixture was concentrated in vacuo to provide the title compound (909 mg, crude) as a brown oil. LC-MS: m / z=632.2 [M+H] + .

[0646] Step B: tert-Butyl (E)-4-(4-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0647] A mixture of tert-butyl (E)-4-(4-(5-carbamoyl-2-((4-(1,3-dioxoisoindolin-2-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate (1.60 g, 2.53 mmol) and hydrazine monohydrate (6.80 mL, 28.0 mmol) in THF (14.0 mL) was stirred at 60° C. for 1 h. After concentration in vacuo, the residue was diluted with 2N aqueous NaOH and then extracted with DCM. The separated organic layer was washed with brine, dried over Na2SO4, filtered and concentrated in vacuo to provide the title compound (1.20 g, crude) as a red foam. LC-MS: m / z=502.2 [M+H] + .

[0648] Step C: tert-Butyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0649] A mixture of tert-butyl (E)-4-(4-(2-((4-aminobut-2-en-1-yl)amino)-5-carbamoyl-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate (1.20 g, 2.39 mmol), methyl 4-chloro-3-methoxy-5-nitrobenzoate (1.18 g, 4.80 mmol) and DIPEA (2.10 mL, 12.06 mmol) in DMF (11 mL) was stirred at 100 °C for 2 h. After quenching the reaction using saturated aqueous NaHCO3 solution, the mixture was extracted twice with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography on NH-SiO2 (DCM:MeOH=50:1) to provide the title compound (770 mg, 43% for three steps) as an orange foam. 1H NMR (400MHz, CDCl3): δ8.42(1H,d,J=1.6Hz),8.12(1H,d,J=2.0Hz),8.10-8.09( 2H,m),7.59(1H,d,J=2.0Hz),7.42(1H,d,J=2.0Hz),5.80-5.67(2H,m),4.73(2H ,d,J=2.0Hz),4.29-4.24(4H,m),3.91(3H,s),3.84(3H,s),3.60(2H,brs),3.17 -3.10(2H,m),2.60(1H,brs),1.76-1.71(2H,m),1.55-1.50(2H,m),1.44(9H,s).

[0650] Step D: tert-Butyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0651] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-4-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitro-phenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=651.2 [M+H] + .

[0652] Step E: Methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-4-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0653] The title compound was prepared in a similar manner to Example 35 (Step E) using tert-butyl (E)-4-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazole-5-carbonylisothiocyanate (Intermediate 3). The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=50:1 to 20:1) to provide the title compound (49% over three steps) as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ7.88-7.78(2H,m),7.65(1H,brs),7.37-7.21(3H,m) ,6.56-6.51(2H,m),5.89-5.82(2H,m),4.91(4H,brs),4.59(2H,brs),4.54(4 H,brs),3.86(3H,s),3.82(2H,brs),3.58(3H,s),2.10-2.08(8H,m),1.99(2H ,brs),1.49-1.46(2H,m),1.34(9H,s),1.28-1.25(7H,m),0.92-0.86(2H,m). LC-MS: m / z=973.3[M+H] + .

[0654] Step F: (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-4-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0655] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-4-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude product was used in the next reaction without purification. LC-MS: m / z=959.5 [M+H] + .

[0656] Step G: tert-Butyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0657] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-4-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by crystallization from hexane / acetone to provide the title compound (54% between two steps) as a pale yellow solid. 1H NMR(400MHz,DMSO-d6):δ7.96-7.90(2H,m),7.64(2H,brs),7.36-7.28(4H,m),6.54-6.52(2H,m),5.94-5.83(2H,m),4.91(4H,brs),4.67(2H,brs), 4.54(4H,brs),3.82(2H,brs),3.68(3H,s),2.10-2.08(7H,m),1.99(2H,b rs),1.49-1.46(2H,m),1.33(9H,s),1.28-1.25(8H,m),0.92-0.86(2H,m). LC-MS:m / z=958.5[M+H] + .

[0658]

[0659] Example 43: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride

[0660] [ka]

[0661] HCl (4M in dioxane 0.800 mL, 3.20 mmol) was added to a solution of tert-butyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate (Example 42, 220 mg, 0.230 mmol) in DCM (10 mL) at 0° C. The reaction mixture was stirred at room temperature for 1 h. After concentration in vacuo, the residue was purified by crystallization from Et2O / MeOH to provide the title compound (190 mg, 93%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6):δ8.78-8.76(1H,m),8.39-8.37(1H,m),8.01-7.99(2H,m),7.65(1H,d,J =0.8Hz),7.64(1H,d,J=0.8Hz),7.41-7.38(3H,m),7.30(1H,s),6.55(1H,s),6.53(1H,s),5.92- 5.81(2H,m),4.91-4.90(4H,m),4.71(2H,s),4.54-4.51(4H,m),3.69(3H,s),3.18-3.15(2H,m) ,2.70-2.67(2H,m),2.10-2.09(6H,m),2.06-2.04(2H,m),1.70-1.67(2H,m),1.29-1.25(8H,m). LC-MS:m / z=858.2[M+H] + .

[0662]

[0663] Example 44: Cyclopropyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0664] [ka]

[0665] The title compound was prepared in a manner similar to Example 6 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and cyclopropyl(4-nitrophenyl)carbonate (Intermediate 7). The crude product was purified by crystallization from hexane / DCM followed by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=100:10:1 to 80:10:1) to provide the title compound (6%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ7.99(1H,s),7.93(1H,s),7.65-7.64(2H,m),7.39-7.38(2H ,m),7.35(1H,s),7.28(1H,s),6.55(1H,s),6.51(1H,s),5.92-5.83(2H,m),4.90(4H, s),4.66(2H,s),4.58-4.50(4H,m),3.93-3.83(2H,m),3.67(3H,s),2.10-2.08(7H,s) ,1.97(2H,brs),1.47(2H,brs),1.32-1.22(8H,m),0.89(2H,brs),0.60-0.52(5H,m). LC-MS:m / z=942.4[M+H] + .

[0666]

[0667] Example 45: 2-Hydroxyethyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0668] [ka]

[0669] Step A: 2-((tert-butyldimethylsilyl)oxy)ethyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0670] The title compound was prepared in a similar manner to Example 10 (Step A) using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carbonate (Intermediate 10). The crude product was purified by crystallization from hexane / Et2O followed by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=100:10:1 to 80:10:1) to provide the title compound (45%) as a white solid. 1 H NMR (400MHz, DMSO-d6): δ7.97(1H,s),7.91(1H,s),7.65-7.64(2H,m),7.36-7.33(2H,m),7.27(1H, s),6.54(1H,s),6.52(1H,s),5.95-5.83(2H,m),4.90-4.89(4H,m),4.64(2H,s),4.53-4.52(4H,m) ,3.98-3.96(2H,m),3.87-3.84(2H,m),3.69-3.67(2H,m),3.65(3H,s),3.50(1H,s),2.10-2.08(7H ,s),1.98(2H,brs),1.52-1.49(2H,m),1.29-1.26(7H,m),1.22(2H,s),0.79(9H,s),-0.02(6H,s).

[0671] Step B: 2-Hydroxyethyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0672] The title compound was prepared in a similar manner to Example 10 (Step B) using 2-((tert-butyldimethylsilyl)oxy)ethyl (E)-4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]-imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo-[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate. The crude product was purified by crystallization from EtO / MeOH to provide the title compound (95%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.98(1H,s),7.92(1H,s),7.66-7.65(2H,m),7.38-7.36(2H ,m),7.28(1H,s),6.56(1H,s),6.52(1H,s),5.95-5.82(2H,m),4.91-4.90(4H,m),4. 66(2H,s),4.56-4.51(4H,m),3.94-3.87(4H,m),3.67(3H,s),2.32(1H,s),2.11-2.0 9(7H,s),2.00-1.98(2H,m),1.52-1.48(2H,m),1.33-1.23(7H,m),0.96-0.86(2H,m). LC-MS:m / z=946.3[M+H] + .

[0673]

[0674] Example 46: tert-Butyl (E)-4-(4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carbonyl)piperidine-1-carboxylate

[0675] [ka]

[0676] The title compound was prepared in a manner similar to Example 12 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=100:10:1 to 70:10:1) to provide the title compound (29%) as a pale yellow solid. 1H NMR(400MHz,DMSO-d6):δ8.00-7.93(2H,m),7.65-7.64(2H,m),7.40-7.29 (4H,m),6.55(1H,s),6.52(1H,s),5.95-5.83(2H,m),4.91-4.90(4H,m),4 .66(2H,brs),4.57-4.50(4H,m),4.27-4.24(1H,m),3.68(3H,s),2.10-2. 08(7H,m),1.98(2H,brs),1.37(9H,s),1.30-1.22(18H,m),1.17(4H,brs). LC-MS: m / z=1069.3[M+H] + .

[0677]

[0678] Example 47: tert-Butyl (E)-4-(4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carbonyl)piperazine-1-carboxylate

[0679] [ka]

[0680] The title compound was prepared in a manner similar to Example 16 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and 1-(tert-butyl)4-(4-nitrophenyl)piperazine-1,4-dicarboxylate (Intermediate 6). The crude product was purified by crystallization from hexane / DCM followed by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=100:10:1 to 80:10:1) to afford the title compound (6%) as a white solid. 1 H NMR(400MHz,DMSO-d6):δ7.98-7.91(2H,m),7.65-7.64(2H,m),7.36-7.29(4H,m), 6.54(1H,s),6.52(1H,s),5.91-5.83(2H,m),4.91(4H,brs),4.67(2H,brs),4.53- 4.52(4H,m),3.68(3H,s),3.50-3.43(6H,m),3.26(4H,brs),2.98(4H,brs),2.10- 2.08(7H,m),1.98-1.97(2H,m),1.48-1.45(2H,m),1.38(9H,s),1.29-1.25(8H,m). LC-MS:m / z=1070.3[M+H] + .

[0681]

[0682] Example 48: (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-((4-(1-(cyclopropylsulfonyl)piperidin-4-yl)but-2-yn-1-yl)oxy)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazole-5-carboxamide

[0683] [ka]

[0684] The title compound was prepared in a manner similar to Example 21 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and cyclopropanesulfonyl chloride. The crude product was purified by crystallization from hexane / DCM followed by column chromatography on SiO2 (DCM:MeOH=20:1 to DCM:MeOH:NH4OH=80:10:1) to provide the title compound (19%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6):δ8.01-7.93(2H,m),7.66-7.64(2H,m),7.39-7.31(4H,m),6.56(1H,s),6.53(1H,s),5.94-5.88(2H,m),4.91( 4H,brs),4.70(2H,brs),4.55-4.53(4H,m),3.71(3H,s),2.10-2.08(7H,m),1.97-1.96(2H,m),1.55-1.53(2H,m),1.30-1.26(7H,m). LC-MS:m / z=962.2[M+H] + .

[0685]

[0686] Example 49: tert-Butyl (E)-4-((4-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidin-1-yl)sulfonyl)piperazine-1-carboxylate

[0687] [ka]

[0688] The title compound was prepared in a manner similar to Example 28 using (E)-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-((4-(piperidin-4-yl)but-2-yn-1-yl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxamide hydrochloride (Example 43) and tert-butyl 4-(chlorosulfonyl)piperazine-1-carboxylate (Intermediate 4). The crude product was purified by crystallization from hexane / DCM followed by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=100:10:1 to 80:10:1) to afford the title compound (44%) as a pale yellow solid. 1H NMR(400MHz,DMSO-d6):δ7.98-7.92(2H,m),7.65-7.64(2H,m),7.36-7.28(4H,m),6.55 (1H,s),6.52(1H,s),5.89-5.86(2H,m),4.90(4H,brs),4.65(2H,brs),4.53-4.52(4H, m),3.67(3H,s),3.02(4H,brs),2.65-2.59(2H,m),2.11-2.09(7H,m),2.01-1.99(2H,m ),1.56-1.53(2H,m),1.39(9H,s),1.29-1.25(8H,m),1.22(4H,brs),1.06-1.00(2H,m). LC-MS:m / z=1106.2[M+H] + .

[0689]

[0690] Example 50: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0691] [ka]

[0692] Step A: tert-Butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0693] The title compound was prepared in a similar manner to Example 1 (Step A) using tert-butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate (Intermediate 24) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (Intermediate 1).

[0694] The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1) to provide the title compound (49%) as a red foam. 1 H NMR(400MHz,DMSO-d6):δ8.15(1H,d,J=1.6Hz),8.06(1H,d,J=1.6Hz),7.93(1H,t,J=6.4Hz),7.73(1H,t,J=6.0Hz),7.53(1H, d,J=1.6Hz),7.31(1H,d,J=1.6Hz),5.56-5.47(2H,m),4.72(2H,s),4.06-4.06(4H,m),3.79(5H,s),3.77(3H,s),1.30(9H,s). LC-MS:m / z=683.2[M+H] + .

[0695]

[0696] Step B: tert-Butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0697] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)azetidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=623.2 [M+H] + .

[0698] Step C: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0699] The title compound was prepared in a similar manner to Example 35 (Step E) using tert-butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)azetidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazole-5-carbonylisothiocyanate (Intermediate 3) in dioxane. The crude solid product was suspended in EtOAc and the solid was filtered off. The filtrate was coagulated from Et2O / hexanes, collected by filtration, washed with hexanes, and dried under vacuum to provide the title compound (18% between two steps) as a light brown solid. 1H NMR(400MHz,DMSO-d6):δ12.90(1H,s),12.84(1H,s),7.86(1H,s),7.77(1H,d,J=1.6Hz),7.67(1 H,d,J=0.8Hz),7.35(1H,s),7.28(1H,s),7.19(1H,d,J=1.2Hz),6.56(1H,s),6.50(1H,s),5.87- 5.84(2H,m),4.91(4H,t,J=6.4Hz),4.57-4.53(6H,m),3.87(3H,s),3.75-3.71(2H,m),3.59(3H, s),3.40-3.35(2H,m),2.33-2.31(2H,m),2.12(6H,d,J=2.4Hz),1.31(9H,s),1.30-1.27(6H,m). LC-MS:m / z=945.3[M+H] + .

[0700] Step D: (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)azetidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0701] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)azetidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was suspended in acetone / hexanes, collected by filtration, washed with hexanes, and dried under vacuum to provide the title compound (87%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ12.84(2H,s),7.92(1H,s),7.74(1H,d,J=1.2Hz),7.67 (1H,s),7.32(2H,s),7.26(1H,d,J=0.8Hz),6.52(2H,d,J=6.8Hz),5.87-5.85(2H ,m),4.91(4H,s),4.62(2H,s),4.53(4H,q,J=6.8Hz),3.73(2H,t,J=8.6Hz),3.6 3(3H,s),2.33(2H,d,J=6.9Hz),2.11(6H,s),1.31(9H,s),1.28(6H,t,J=7.2Hz). LC-MS: m / z=931.3[M+H] + .

[0702] Step E: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)azetidine-1-carboxylate

[0703] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)azetidin-3-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude solid compound was suspended in water and collected by filtration. The crude solid was purified by recrystallization from acetone / Et2O, collected by filtration, and dried under vacuum to provide the title compound (62%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ12.79(2H,s),7.92(1H,s),7.86(1H,s),7.62(2H,d,J=5.2 Hz),7.30(3H,s),7.24(1H,s),6.49(2H,d,J=4.0Hz),5.85-5.82(2H,m),4.87(4H,s) ,4.62(2H,s),4.49(4H,d,J=6.8Hz),3.70(2H,t,J=8.2Hz),3.63(3H,s),3.37-3.35( 2H,m),2.29-2.28(2H,m),2.06(6H,d,J=3.2Hz),1.27(9H,s),1.24(6H,t,J=7.3Hz). LC-MS:m / z=930.3[M+H] + .

[0704]

[0705] Example 51: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0706] [ka]

[0707] Step A: tert-Butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0708] The title compound was prepared in a similar manner to Example 1 (Step A) using tert-butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate (Intermediate 25) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate hydrochloride (Intermediate 1). The crude product was purified by column chromatography on NH-SiO2 (DCM only to DCM:MeOH=98:2) to provide the title compound (83%) as an orange solid. LC-MS: m / z=697.2 [M+H] + .

[0709] Step B: tert-Butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0710] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown solid. LC-MS: m / z=637.2 [M+H] + .

[0711] Step C: Methyl 2-((amino-13-bromanilidene)amino)-1-((2E)-4-(2-((amino-13-bromanilidene)amino)-7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0712] The title compound was prepared in a similar manner to Example 1 (Step C) using tert-butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate. The crude solid product was recrystallized from Et2O / MeOH, collected by filtration, washed with Et2O, and dried under vacuum to provide the title compound (52%) as a yellow solid. LC-MS: m / z=687.2 [M+H] + .

[0713] Step D: Methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0714] The title compound was prepared in a similar manner to Example 1 (Step D) using methyl (E)-2-amino-1-(4-(2-amino-7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-7-methoxy-1H-benzo[d]imidazole-5-carboxylate dihydrobromide and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid. The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=95:5:1) to provide the title compound (15%) as a purple solid. 1 H NMR(400MHz,DMSO-d6):δ12.89(1H,s),12.83(1H,s),7.86(1H,s),7.76(1H,s),7.67(1H,s),7.35(1H,s),7.28(1H,d,J=12.0Hz) ,7.17(1H,d,J=16.8Hz),6.55(1H,s),6.51(1H,s),5.93-5.79(2H,m),4.91(4H,d,J=4.0Hz),4.57-4.50(6H,m),3.86(3H,s),3.59 and 3.52(3H,s+s),3.29-3.24(1H,m),3.20-3.13(1H,m),3.06-2.98(1H,m),2.76-2.73(1H,m),2. 14(2H,s),2.10(7H,s),1.74-1.73(1H,m),1.35(10H,d,J=2.8Hz),1.29(6H,td,J=7.1,2.5Hz).

[0715] Step E: (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0716] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was washed with water and dried under vacuum to provide the title compound (69%) as a purple solid. 1 H NMR(400MHz,DMSO-d6):δ12.83(2H,d,J=10.8Hz),7.87(1H,s),7.72(1H,s),7.62(1H,s),7.3 2-7.29(2H,m),7.22(1H,d,J=12.0Hz),6.50(1H,s),6.47(1H,s),5.88-5.78(2H,m),4.88(4H, s),4.62-4.56(2H,m),4.50(4H,d,J=6.8Hz),3.62-3.56(3H,s+s),3.03-2.93(1H,m),2.71(1 H,t,J=8.8Hz),2.11(2H,s),2.06(7H,s),1.70(1H,brs),1.30(10H,s),1.24(6H,t,J=7.0Hz).

[0717] Step F: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)pyrrolidine-1-carboxylate

[0718] The title compound was prepared in a similar manner to Example 3 using (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)but-2-yn-1-yl)oxy)-5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude product was purified by column chromatography on NH-SiO2 (DCM:MeOH=93:7) to provide the title compound (31%) as a red-brown solid. 1 H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.93(1H,s),7.87(1H,s),7.67(1H,s),7.65(1H,s) ,7.30-7.24(4H,m),6.51(2H,s),5.93-5.79(2H,m),4.90(4H,s),4.64(2H,d,J=22.0Hz),4 .56-4.53(4H,m),3.67-3.62(3H,s+s),3.25-3.17(1H,m),3.05(1H,t,J=8.0Hz),2.77(1H, t,J=8.8Hz),2.16(2H,s),2.10(7H,s),1.75(1H,brs),1.34(10H,s),1.27(6H,t,J=6.4Hz). LC-MS:m / z=944.3[M+H] + .

[0719]

[0720] Example 52: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0721] [ka]

[0722] Step A: tert-Butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0723] The title compound was prepared in a similar manner to Example 1 (Step A) using tert-butyl 3-(4-(5-carbamoyl-2-chloro-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate (Intermediate 26) and methyl (E)-4-((4-aminobut-2-en-1-yl)amino)-3-methoxy-5-nitrobenzoate (Intermediate 1). The crude product was purified by column chromatography on SiO2 (DCM only to DCM:MeOH:NH4OH=200:10:1) to provide the title compound (41%) as a red foam. 1 H NMR(400MHz,DMSO-d6):δ8.15(1H,d,J=2.0Hz),8.07(1H,d,J=2.0Hz),7.94(1H,t,J=6.2Hz ),7.72(1H,t,J=5.6Hz),7.56(1H,d,J=1.6Hz),7.32(1H,d,J=1.6Hz),5.56-5.55(2H,m),4 .75(2H,s),4.06(4H,s),3.79(3H,s),3.77(3H,s),3.67(1H,d,J=12.8Hz),2.63-2.57(1H, m),2.12(2H,t,J=5.6Hz),1.66-1.64(1H,m),1.48-1.44(2H,m),1.32(9H,s),1.19(1H,s). LC-MS:m / z=711.2[M+H] + .

[0724] Step B: tert-Butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0725] The title compound was prepared in a similar manner to Example 1 (Step B) using tert-butyl (E)-3-(4-(5-carbamoyl-2-((4-((2-methoxy-4-(methoxycarbonyl)-6-nitrophenyl)amino)but-2-en-1-yl)amino)-3-nitrophenoxy)but-2-yn-1-yl)piperidine-1-carboxylate. The crude product was used in the next reaction without purification as a light brown foam. LC-MS: m / z=651.2 [M+H] + .

[0726] Step C: Methyl (E)-1-(4-(7-((3-(1-(tert-butoxycarbonyl)piperidin-3-yl)prop-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate

[0727] The title compound was prepared in a similar manner to Example 35 (Step E) using tert-butyl (E)-3-(4-(3-amino-2-((4-((2-amino-6-methoxy-4-(methoxycarbonyl)phenyl)amino)but-2-en-1-yl)amino)-5-carbamoylphenoxy)but-2-yn-1-yl)piperidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazole-5-carbonylisothiocyanate (Intermediate 3). The crude product was purified by column chromatography on SiO2 (DCM:MeOH:NH4OH=200:10:1) to provide the title compound (54% between two steps) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ12.86(1H,s),12.80(1H,s),7.84(1H,s),7.74(1H,s),7.63(1H,s),7 .32(1H,s),7.27(1H,s),7.18(1H,s),6.52(1H,s),6.46(1H,s),5.82-5.81(2H,m),4.89(4H,d, J=14.8Hz),4.56-4.49(6H,m),3.83(3H,s),3.65-3.62(1H,m),3.55(3H,s),2.07(6H,s),1.97 -1.93(2H,m),1.45-1.60(1H,m),1.37(2H,s),1.29(9H,s),1.24(6H,t,J=6.6Hz),1.19(1H,s). LC-MS:m / z=973.4[M+H] + .

[0728] Step D: (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid

[0729] The title compound was prepared in a similar manner to Example 1 (Step E) using methyl (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylate. The crude solid product was suspended in acetone / hexanes, collected by filtration, washed with hexanes, and dried under vacuum to provide the title compound (93%) as a white solid. 1H NMR(400MHz,DMSO-d6):δ12.83(2H,s),7.94(1H,s),7.74(1H,d,J=1.2Hz),7.67(1H,s),7.33(2H ,s),7.27(1H,d,J=0.8Hz),6.53(2H,d,J=5.2Hz),5.87-5.82(2H,m),4.92(4H,d,J=16.0Hz),4.63 (2H,s),4.54(4H,q,J=6.8Hz),3.67(1H,d,J=14.6Hz),3.62(3H,s),2.12(6H,d,J=2.8Hz),1.99-1 .97(2H,m),1.57-1.57(1H,m),1.41(1H,d,J=12.8Hz),1.32(10H,s),1.28(7H,td,J=7.1,1.3Hz). LC-MS:m / z=959.3[M+H] + .

[0730] Step E: tert-Butyl (E)-3-(4-((5-carbamoyl-1-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-methoxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)but-2-yn-1-yl)piperidine-1-carboxylate

[0731] The title compound was prepared in a manner similar to Example 3 using (E)-1-(4-(7-((4-(1-(tert-butoxycarbonyl)piperidin-3-yl)but-2-yn-1-yl)oxy)-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-methoxy-1H-benzo[d]imidazole-5-carboxylic acid. The crude solid product was suspended in water and collected by filtration. The solid was purified by recrystallization from acetone / Et2O, collected by filtration, and dried under vacuum to provide the title compound (70%) as a light brown solid. 1H NMR (400MHz, DMSO-d6): δ12.84(2H,s),7.96(1H,s),7.90(1H,s),7.67-7.65(2H,m),7. 36(3H,s),7.29(1H,s),6.53(2H,d,J=3.6Hz),5.89-5.86(2H,m),4.93(4H,d,J=19.2Hz ),4.67(2H,s),4.54(4H,q,J=7.0Hz),3.67(4H,s),2.10(6H,d,J=4.8Hz),1.99-1.97(2 H,m),1.57(1H,s),1.42-1.40(1H,m),1.32(9H,s),1.28(7H,t,J=7.2Hz),1.23(1H,s). LC-MS:m / z=958.3[M+H] + .

[0732]

[0733] Biological activity A:50nM, B:50~500nM, C:500nM, ultra

[0734]

Table 1

[0735]

[0736] Cell strain および trial

[0737] THP1-Blue cells (catalog number thp-isg) were purchased from InvivoGen (San Diego, USA). Penicillin-Streptomycin (catalog number L0022-100), Fetal Bovine Serum (FBS) (catalog number S1480-500) and Roswell Park Memorial Institute (RPMI) 1640 medium (catalog number L0498-500) were purchased from biowest (Noyet, France). Zeocin (catalog number ant-zn-05) and Normocin (catalog number ant-nr-1) were purchased from InvivoGen (San Diego, USA). QUANTI-Blue™ (catalog number rep-qbs) was purchased from InvivoGen. THP1-blue cells were cultured in RPMI 1640 medium with 10% heat-inactivated FBS, 100 μg / ml normocin, 100 μg / ml zeocin, and 100 U / ml to 100 μg / ml penicillin-streptomycin. Cells were cultured at 37°C in a humidified 5% CO2 atmosphere.

[0738] ISG reporter analysis

[0739] 5x10 THP1-blue cells 4 Cells were seeded at 90 μl / well. 10 μl of compound was added to the cells and incubated for 24 hours. 20 μl of cell culture supernatant was added to 180 μl of QUANTI-Blue. TM The solution was dispensed into a new 96-well plate (cat. no. 30096) and incubated for 15 min at 37° C. The absorbance was measured at 655 nm by Varioskan Lux (Thermofisher Scientific, USA). [Industrial Applicability]

[0740] The compounds of the present invention can be used as agonists of stimulator of interferon genes (STING) and are useful in the treatment of cancer and certain infectious diseases.

Claims

1. Compounds of Formula I: [Chemical formula I] 【Chemistry 1】 [In the formula, Each R １ is independently NH ２ , O.H., N.H.O.R. ４ or N.H.R. ４ and Each R ２ are independently hydrogen, halogen, CF ３ , -(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkyl, acyl, amino, substituted amino, cyano, acyloxy or aryloxy; R ３ is hydrogen, halogen, CF ３ , acyl, amino, substituted amino, -(C １ -C ６ ) alkyl, -(C １ -C ６ ) haloalkyl, -(C ３ -C ６ ) cycloalkyl, -(C １ -C ６ ) alkoxy, -(C １ -C ６ ) Hydroxyalkyl, hetCyc １ , hetCyc ２ , -(C １ -C ３ ) alkyl [hetCyc １ ], -(C １ -C ３ ) alkyl [hetCyc ２ ], -(C １ -C ３ ) alkyl [hetAr ２ ], -(C １ -C ３ ) alkyl [hetAr ３ ], cyano, nitro, alkoxy, acyloxy or aryloxy; R ４ is H, trifluoromethyl, -(C １ -C ６ ) alkyl, -(C ２ -C ６ ) alkenyl, -(C ２ -C ６ ) alkynyl, -(C １ -C ３ ) alkyl [(C ３ -C ６ ) cycloalkyl], -(C １ -C ６ ) fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, -(C １ -C ６ ) trifluoroalkyl, -(C １ -C ６ ) alkylamine, -(C １ -C ６ ) hydroxyalkyl, -(C ２ -C ６ ) dihydroxyalkyl, [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl- or [(C １ -C ６ ) alkoxy]-[(C １ -C ６ ) alkoxy]-(C １ -C ６ ) alkyl-; Each X １ and X ２ are independently N or CR ５ and R ５ is hydrogen, halogen, OH, CF ３ , acyl, amino, substituted amino, -(C １ -C ６ ) alkyl, substituted (C １ -C ６ ) alkyl, -(C １ -C ６ ) haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) hydroxyalkyl, —O(C ２ -C ６ ) alkenyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [hetCyc １ ], -O(C １ -C ６ ) alkyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkenyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkynyl [(C １ -C ６ )alkoxy], —O(C １ -C ６ ) alkyl [(C ３ -C ６ )cycloalkyl], —O(C １ -C ３ ) alkyl [hetCyc １ ], -O(C １ -C ３ ) alkyl [hetCyc ２ ] or -O(C ２ -C ６ ) alkynyl [hetCyc １ ]; hetCyc １ is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being -(C １ -C ６ ) alkyl, -(C １ -C ６ ) alkoxy, OH, halogen, -C(O)R ６ , -CO ２ R ６ , -C(O)NR ６ R ７ , -S(O) ２ N.R. ６ R ７ Or -S(O) ２ R ６ is optionally substituted with a substituent selected from hetCyc ２ is a bridged 8-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom; R ６ is H, -(C １ -C ６ ) alkyl, -(C １ -C ６ ) fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, -(C １ -C ６ ) trifluoroalkyl, -(C １ -C ６ alkyl)-NR ７ R ８ , -(C １ -C ６ ) hydroxyalkyl, -(C ２ -C ６ ) dihydroxyalkyl, [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl-, [(C １ -C ６ ) alkoxy]-[(C １ -C ６ ) alkoxy]-(C １ -C ６ ) alkyl-, hetCyc １ , Ar １ , hetAr ２ or hetAr ３ and R ７ is H or -(C １ -C ６ ) alkyl; N.R. ６ R ７ forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being -(C １ -C ６ ) Alkyl, OH, NH ２ , -(C １ -C ６ ) hydroxyalkyl, -(C １ -C ６ ) alkylamine, —CO ２ R ８ and -(C １ -C ３ ) Alkyl CO ２ R ８ is optionally substituted with one or more substituents independently selected from R ８ is H, -(C １ -C ３ ) alkyl or -(C １ -C ３ ) hydroxyalkyl; Ar １ (C １ -C ６ ) alkoxy, halogen, (C １ -C ６ ) alkyl and CF ３ phenyl optionally substituted with one or more substituents independently selected from hetAr ２ is a halogen, CF ３ , (C １ -C ６ ) alkyl and (C １ -C ６ ) pyridyl optionally substituted with one or more substituents independently selected from alkoxy; hetAr ３ has 2 to 3 ring heteroatoms independently selected from N, O and S, (C １ -C ６ ) 5-membered heteroaryl, optionally substituted with alkyl and OH; Y １ and Y ２ are each independently N, O, S, or CR. ９ , N.R. １０ and R ９ is hydrogen, halogen, CF ３ , -(C １ -C ６ ) alkyl or -O(C １ -C ６ ) alkyl; R １０ is hydrogen, -(C １ -C ６ ) alkyl or acyl; A is -CH=CH-, -CC-, -CH ２ -, -CR １１ R １２ --, --C(O)NR １３ -, -C(O)NHOR １４ --, --NR １３ C(O)-, -NR １３ CO ２ --, --NR １３ C(O)NR １３ --, --NR １３ -, -(C ３ -C ７ ) cycloalkyl-, -O-, -S-, -S(O)- or -S(O) ２ -, optionally substituted -phenyl-, optionally substituted -(5- to 6-membered heteroaryl)- or optionally substituted -(5- to 6-membered heterocycloalkyl)-; R １１ F, CF ３ , -(C １ -C ６ ) alkyl, substituted (C １ -C ６ ) alkyl, cyano; R １２ H, F, CF ３ , -(C １ -C ６ ) alkyl; R １１ and R １２ form together with the atom(s) to which they are attached a 3- to 7-membered carbocyclic or heterocyclic ring; R １３ is hydrogen, -(C １ -C ６ ) C alkyl, -(C １ -C ６ )C fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, or (C １ -C ６ ) trifluoroalkyl; R １４ (C １ -C ６ ) alkyl, (C １ -C ６ ) fluoroalkyl, (C １ -C ６ ) difluoroalkyl, (C １ -C ６ ) trifluoroalkyl or [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl-; m is 0, 1, 2 or 3; n is 0, 1, 2 or 3.

2. The compound of claim 1, wherein formula I includes compounds of formula II-a, II-b, and II-c: [Chemical formula II-a] 【Chemistry 2】 [Chemical formula II-b] 【Chemistry 3】 [Chemical formula II-c] 【Chemistry 4】 [Each R ２ are independently hydrogen, halogen, CF ３ , -(C １ -C ６ ) alkyl, —O(C １ -C ６ ) alkyl, acyl, amino, substituted amino, cyano, acyloxy or aryloxy; R ３ is hydrogen, halogen, CF ３ , acyl, amino, substituted amino, -(C １ -C ６ ) alkyl, -(C １ -C ６ ) haloalkyl, -(C ３ -C ６ ) cycloalkyl, -(C １ -C ６ ) alkoxy, -(C １ -C ６ ) Hydroxyalkyl, hetCyc １ , hetCyc ２ , -(C １ -C ３ ) alkyl [hetCyc １ ], -(C １ -C ３ ) alkyl [hetCyc ２ ], -(C １ -C ３ ) alkyl [hetAr ２ ], -(C １ -C ３ ) alkyl [hetAr ３ ], cyano, nitro, alkoxy, acyloxy or aryloxy; hetCyc １ is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being -(C １ -C ６ ) alkyl, -(C １ -C ６ ) alkoxy, OH, halogen, -C(O)R ６ , -CO ２ R ６ , -C(O)NR ６ R ７ , -S(O) ２ N.R. ６ R ７ Or -S(O) ２ R ６ is optionally substituted with a substituent selected from hetCyc ２ is a bridged 8-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom; Each X １ and X ２ are independently N or CR ５ and R ５ is hydrogen, halogen, OH, CF ３ , acyl, amino, substituted amino, -(C １ -C ６ ) alkyl, substituted (C １ -C ６ ) alkyl, -(C １ -C ６ ) haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) hydroxyalkyl, —O(C ２ -C ６ ) alkenyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [hetCyc １ ], -O(C １ -C ６ ) alkyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkenyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkynyl [(C １ -C ６ )alkoxy], —O(C １ -C ６ ) alkyl [(C ３ -C ６ )cycloalkyl], —O(C １ -C ３ ) alkyl [hetCyc １ ], -O(C １ -C ３ ) alkyl [hetCyc ２ ], or -O(C ２ -C ６ ) alkynyl [hetCyc １ ]; R ６ is H, -(C １ -C ６ ) alkyl, -(C １ -C ６ ) fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, -(C １ -C ６ ) trifluoroalkyl, -(C １ -C ６ alkyl)-NR ７ R ８ , -(C １ -C ６ ) hydroxyalkyl, -(C ２ -C ６ ) dihydroxyalkyl, [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl-, [(C １ -C ６ ) alkoxy]-[(C １ -C ６ ) alkoxy]-(C １ -C ６ ) alkyl-, hetCyc １ , Ar １ , hetAr ２ or hetAr ３ and R ７ is H or -(C １ -C ６ ) alkyl; N.R. ６ R ７ forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being -(C １ -C ６ ) Alkyl, OH, NH ２ , -(C １ -C ６ ) hydroxyalkyl, -(C １ -C ６ ) alkylamine, —CO ２ R ８ and -(C １ -C ３ ) Alkyl CO ２ R ８ optionally substituted with one or more substituents independently selected from R ８ is H, -(C １ -C ３ ) alkyl or -(C １ -C ３ ) hydroxyalkyl; Ar １ (C １ -C ６ ) alkoxy, halogen, (C １ -C ６ ) alkyl and CF ３ phenyl optionally substituted with one or more substituents independently selected from hetAr ２ is halogen, CF ３ , (C １ -C ６ ) alkyl and (C １ -C ６ ) pyridyl optionally substituted with one or more substituents independently selected from alkoxy; hetAr ３ has 2 to 3 ring heteroatoms independently selected from N, O and S, (C １ -C ６ ) 5-membered heteroaryl, optionally substituted with alkyl and OH; Y １ and Y ２ are each independently N, O, S, or CR. ９ , N.R. １０ and R ９ is hydrogen, halogen, CF ３ , -(C １ -C ６ ) alkyl or -O(C １ -C ６ ) alkyl; R １０ is hydrogen, -(C １ -C ６ ) alkyl or acyl.

3. The compound of claim 1, wherein formula I includes compounds of formula III-a, III-b, III-c, and III-d: [Formula III-a]. 【Chemistry 5】 [Chemical formula III-b] 【Chemistry 6】 [Chemical formula III-c] 【Chemistry 7】 [Chemical formula III-d] 【Chemistry 8】 [Z is -R ６ , -OR ６ , -NR ６ R ７ , -(C １ -C ６ ) alkyl, -(C １ -C ６ ) fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, -(C １ -C ６ ) trifluoroalkyl, -(C １ -C ６ alkyl)-NR ７ R ８ , -(C １ -C ６ ) hydroxyalkyl, -(C ２ -C ６ ) dihydroxyalkyl, [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl- or [(C １ -C ６ ) alkoxy]-[(C １ -C ６ ) alkoxy]-(C １ -C ６ ) alkyl-; L is CH or N; Each X １ and X ２ are independently N or CR ５ and R ５ is hydrogen, halogen, OH, CF ３ , acyl, amino, substituted amino, -(C １ -C ６ ) alkyl, substituted (C １ -C ６ ) alkyl, -(C １ -C ６ ) haloalkyl, cyano, nitro, alkoxy, acyloxy, aryloxy, -O(C １ -C ６ ) alkyl, —O(C １ -C ６ ) hydroxyalkyl, —O(C ２ -C ６ ) alkenyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [(C ４ -C ６ ) hydroxy], —O(C ２ -C ６ ) alkynyl [hetCyc １ ], -O(C １ -C ６ ) alkyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkenyl [(C １ -C ６ )alkoxy], —O(C ２ -C ６ ) alkynyl [(C １ -C ６ )alkoxy], —O(C １ -C ６ ) alkyl [(C ３ -C ６ )cycloalkyl], —O(C １ -C ３ ) alkyl [hetCyc １ ], -O(C １ -C ３ ) alkyl [hetCyc ２ ], or -O(C ２ -C ６ ) alkynyl [hetCyc １ ]; R ６ is H, -(C １ -C ６ ) alkyl, -(C １ -C ６ ) fluoroalkyl, -(C １ -C ６ ) difluoroalkyl, -(C １ -C ６ ) trifluoroalkyl, -(C １ -C ６ alkyl)-NR ７ R ８ , -(C １ -C ６ ) hydroxyalkyl, -(C ２ -C ６ ) dihydroxyalkyl, [(C １ -C ６ ) alkoxy](C １ -C ６ ) alkyl-, [(C １ -C ６ ) alkoxy]-[(C １ -C ６ ) alkoxy]-(C １ -C ６ ) alkyl-, hetCyc １ , Ar １ , hetAr ２ or hetAr ３ and R ７ is H or -(C １ -C ６ ) alkyl; N.R. ６ R ７ forms a 4-6 membered ring having one or more heteroatoms selected from N and O, said ring being -(C １ -C ６ ) Alkyl, OH, NH ２ , -(C １ -C ６ ) hydroxyalkyl, -(C １ -C ６ ) alkylamine, —CO ２ R ８ and -(C １ -C ３ ) Alkyl CO ２ R ８ optionally substituted with one or more substituents independently selected from R ８ is H, -(C １ -C ３ ) alkyl or -(C １ -C ３ ) hydroxyalkyl; hetCyc １ is a 4-6 membered heterocyclic ring containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur, said heterocyclic ring being -(C １ -C ６ ) alkyl, -(C １ -C ６ ) alkoxy, OH, halogen, -C(O)R ６ , -CO ２ R ６ , -C(O)NR ６ R ７ , -S(O) ２ N.R. ６ R ７ Or -S(O) ２ R ６ is optionally substituted with a substituent selected from hetCyc ２ is a bridged 8-membered heterocyclic ring having an annular nitrogen atom, and optionally an annular oxygen atom; Ar １ (C １ -C ６ ) alkoxy, halogen, (C １ -C ６ ) alkyl and CF ３ phenyl optionally substituted with one or more substituents independently selected from hetAr ２ is halogen, CF ３ , (C １ -C ６ ) alkyl and (C １ -C ６ ) pyridyl optionally substituted with one or more substituents independently selected from alkoxy; hetAr ３ has 2 to 3 ring heteroatoms independently selected from N, O and S, (C １ -C ６ ) is a 5-membered heteroaryl optionally substituted with alkyl and OH.

4. A pharmaceutical composition comprising a pharma- ceutically effective amount of a compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt, solvate, polymorph, ester, tautomer or prodrug thereof, and a pharma- ceutically acceptable carrier.

5. 11. A method of stimulating interferon gene expression in a human patient, comprising administering to said patient an effective amount of a compound according to any one of claims 1 to 3 or a pharma- ceutically acceptable salt thereof.

6. 13. A method of treating a tumor in a patient comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 3 or a pharma- ceutically acceptable salt thereof.

7. A method of treating a STING-mediated disease or disorder in a human in need thereof, comprising administering to an individual in need thereof an effective amount of the composition described in claim 4 or a pharma- ceutically acceptable salt thereof.

8. 8. The method of claim 7, wherein the proliferative disorder is selected from the group consisting of cancer diseases and certain infectious diseases.

9. 9. The method of claim 8, wherein the disease or disorder is selected from influenza, HIV, HCV, HPV or HBV infection.

10. 5. The use of a compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof according to claim 4, for use as a vaccine adjuvant.

11. 7. The method of claim 5 or claim 6, wherein the administering step comprises intravenous or intratumoral administration, or both.

12. The method of claim 5 or claim 6, wherein the administering step comprises administering the compound to the patient as an antibody-drug conjugate or in a liposomal formulation.

13. 7. The method of claim 5 or claim 6, further comprising administering an effective amount of an immune checkpoint targeted drug.

14. The method of claim 12, wherein the immune checkpoint targeting drug comprises an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, or an anti-4-1BB antibody.

15. 7. The method of claim 5 or claim 6, further comprising administering an effective amount of a chemotherapeutic agent.

16. 7. The method of claim 5 or claim 6, further comprising administering an effective amount of a small molecule kinase-targeted drug.

17. 7. The method of claim 5 or claim 6, further comprising administering ionizing radiation or an anti-cancer drug.