STING agonists containing hydrazide, hydrazine, and hydroxamic acid functional groups

Abstract

The compounds disclosed herein are prepared for use in the treatment of a disease, disorder, or condition treatable by activation of the stimulator of interferon genes (STING) adaptor protein, for example, the treatment or prevention of cancer. [Formula 1] JPEG2026508759000428.jpg48170(I)

Description

[Technical Field]

[0001] The present invention relates to novel compounds that are Stimulator of Interferon Genes (STING) agonists and are useful in the treatment of disorders affected by STING protein regulation. The invention also relates to pharmaceutical compositions containing one or more such compounds, processes for preparing such compounds and compositions, and the use of such compounds or pharmaceutical compositions to treat various diseases, syndromes, and disorders affected by STING protein regulation, for example, in the treatment of several different cancers. [Background technology]

[0002] Stimulator of interferon genes (STING, also known as transmembrane protein 173 / TMEM173 / MPYS / MITA / ERIS) is a signaling molecule encoded by the TMEM173 gene in humans. STING is a 379-amino acid protein consisting of several transmembrane domains. STING protein is expressed in several endothelial and epithelial cell types as well as in hematopoietic lineages, which may include or exclude T cells, dendritic cells (DCs), including plasmacytoid dendritic cells (pDCs), and macrophages. STING associates with the endoplasmic reticulum (ER) within cells and plays a key role in regulating the transcription of numerous host defense genes, including type I interferons (IFNs) and pro-inflammatory cytokines.

[0003] Recognition of aberrant DNA species or cyclic dinucleotides (CDNs) in the cytosol of cells leads to activation of STING. Cytosolic DNA species can activate STING signaling after binding to cyclic GMP-AMP synthase (cGAS). Binding of cytosolic DNA to cGAS catalyzes the production of a type of CDN known as cyclic GMP-AMP (cGAMP), which contains one 2',5'-phosphodiester bond and a canonical 3',5' linkage (c[G(2',5')pA(3',5')p]). Binding of cGAMP and other bacterial CDNs induces a conformational change in the STING protein, promoting the binding of TANK-binding kinase 1 (TBK1). The STING-TBK1 complex further translocates to the perinuclear region of the cell and transports TBK1 to the endolysosomal compartment, where it phosphorylates transcription factors such as interferon regulatory factor 3 (IRF3). Similarly, STAT6 and nuclear factor-κB (NF-κB) are also activated downstream of STING activation. These transcription factors then translocate into the nucleus to initiate innate immune gene transcription and the production of type I IFN and other cytokines. STING is then rapidly degraded, an event that may avoid problems associated with sustained cytokine production (Non-Patent Document 1).

[0004] Recent evidence supports the finding that activation of STING by CDNs within the tumor microenvironment, preferably in tumor-resident dendritic cells, promotes the release of type I IFN and TNFα, thereby resulting in immune-mediated antitumor responses. STING-dependent activation of antigen-presenting cells (APCs) efficiently drives highly specific T cell priming against neoantigens (Non-Patent Document 2). STING activation not only leads to the generation of tumor-specific killer T cells that directly eradicate tumors, but also to vaccine-like long-term immune responses that protect against cancer recurrence.

[0005] Studies in mice have shown that type I IFN signaling plays an important role in tumor-initiating T cell priming and tumor control (Non-Patent Document 3). Mice lacking IFN-α / β receptors in DCs are unable to reject immunogenic tumors, and CD8α-positive DCs derived from these mice are defective in antigen cross-presentation to CD8-positive T cells. Furthermore, transcriptional profiling analysis of melanoma patients revealed that tumors containing infiltrating activated T cells are characterized by a type I IFN transcriptional signature (Non-Patent Document 4).

[0006] Therefore, synthetic STING agonists are particularly interesting as potential anticancer agents. Synthetic STING agonists are generally disclosed, for example, in PCT Publication Nos. WO 2017 / 175147, WO 2017 / 175156, WO 2019 / 069269, WO 2019 / 069270, and WO 2019 / 069275. Activation or inhibition of type I interferon production is an important strategy for treating or preventing human diseases, including viral infections and autoimmune diseases. It has been found that compounds that activate or inhibit type I interferon production may be useful not only in infectious diseases and innate immunity, but also in cancer (Non-Patent Document 5), allergic diseases (Non-Patent Document 6), neurodegenerative diseases such as amyotrophic lateral sclerosis and multiple sclerosis (Non-Patent Document 7, Non-Patent Document 8, Non-Patent Document 9), other inflammatory conditions such as irritable bowel disease (Non-Patent Document 10), and as vaccine adjuvants (Non-Patent Document 11, Non-Patent Document 12).

[0007] STING is essential for antibacterial host defense, including defense against various DNA and RNA viruses and bacteria (reviewed in Non-Patent Document 13 and Non-Patent Document 14). The Herpesviridae, Flaviviridae, Coronaviridae, Papillomaviridae, Adenoviridae, Hepadnaviridae, Orthomyxoviridae, Paramyxoviridae, and Rhabdoviridae families have evolved mechanisms to inhibit STING-mediated type I interferon production and evade host immune control (Non-Patent Document 15, Non-Patent Document 16, Non-Patent Document 17, Non-Patent Document 18, Non-Patent Document 19, Non-Patent Document 20, Non-Patent Document 21, Non-Patent Document 22, Non-Patent Document 23, Non-Patent Document 24, Non-Patent Document 25). Therefore, small molecule activation of STING may be beneficial for the treatment of these infectious diseases.

[0008] In contrast, increased and prolonged type I IFN production is associated with various chronic infections, including mycobacteria (Non-Patent Document 26, Non-Patent Document 27, Non-Patent Document 28), Francisella (Non-Patent Document 29, Non-Patent Document 30), Chlamydia (Non-Patent Document 31), Plasmodium (Non-Patent Document 32), and HIV (Non-Patent Document 33, Non-Patent Document 34). Similarly, excessive type I interferon production is found in patients with complex forms of autoimmune disease. Support from genetic evidence in humans and studies in animal models supports the hypothesis that inhibition of STING results in a reduction of type I interferon, which drives autoimmune disease (Non-Patent Document 35, Non-Patent Document 36). Therefore, inhibitors of STING offer a therapeutic approach for patients with chronic type I interferon and pro-inflammatory cytokine production associated with infections or complex autoimmune diseases. Allergic diseases are associated with a Th2-biased immune response to allergens. Th2 responses are associated with elevated levels of IgE, which, through its effects on mast cells, promotes hypersensitivity to allergens, resulting in the symptoms seen in, for example, allergic rhinitis and asthma. In healthy individuals, the immune response to allergens is more balanced, with a mixed Th2 / Th1 response and regulatory T cell response. Induction of type 1 interferon has been shown to reduce Th2-type cytokines in the local environment and promote Th1 / Treg responses. In this context, induction of type 1 interferon, for example, by activation of STING, may be beneficial in the treatment of allergic diseases such as asthma and allergic rhinitis (Non-Patent Document 37).

[0009] In view of the above, compounds that modulate STING are useful for treating one or more diseases selected from the group consisting of inflammatory, allergic, and autoimmune diseases, infectious diseases, cancer, and precancerous syndromes, and / or as immunogenic compositions or vaccine adjuvants. Of particular relevance is the immunotherapy of cancer and viral infections, particularly prostate cancer, renal cell carcinoma, melanoma, pancreatic cancer, cervical cancer, ovarian cancer, colon cancer, head and neck cancer, lung cancer, fibrosarcoma, breast cancer, and hepatitis B. [Preliminary Technology Documents] [License]

[0010] [License 1] PCT International Publication No. 2017 / 175147 [License 2] PCT International Publication No. 2017 / 175156 [License 3] PCT International Publication No. 2019 / 069269 [License 4] PCT International Publication No. 2019 / 069270 [Patent Document 5] PCT International Publication No. 2019 / 069275 [Non-licensed literature]

[0011] [Non-licensed Document 1] Nature Reviews Immunol, 2015, 15:760-770;Cell Reports, 2015, 11:1018-1030 [Non-licensed Document 2] L. Corrales and TF.Gajewski, Clin Cancer Res, 2015, 21(21),pp. 4774-9 [Non-licensed Document 3] J. Exp. Med. 2011, 208, 1989-2003 [Non-licensed Document 4] Cancer Res. 2009, 69:3077-3085 [Non-licensed Document 5] L. Zitvogel et al.,Nature Reviews Immunology, 2015, vol. 15(7),pp. 405-414 [Non-licensed Document 6] J. Moisan et al.,Am. J. Physiol. Lung Cell Mol. Physiol. ,2006, vol. 290, L987-995

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[0012] Therefore, there is a need for compounds that modulate the activity of STING, thus providing therapeutic benefit in the treatment of diseases in which modulation of STING is beneficial. [Means for solving the problem]

[0013] In one aspect, the present disclosure provides a compound of formula I: [ka] (I) or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: Ring A is [ka] (a) (b) (c) (d) is selected from the group consisting of During the ceremony, G and G1 are independently N, CH, or C-X1-R2; G' and G2 are independently N or CH; X is NR, O, or S; X' is N or CH; X1 is CH2, O or S; R is hydrogen or C 1-4 is alkyl, When G and G1 are each C-X1-R2, the R2 groups may be linked to form L1; L and L1 are each independently C 2-4 Alkylene or C 2-4 is alkenylene, R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is -OH or -NR9R 10 and R9 and R 10 is independently selected from hydrogen and C1-C6 alkyl; X2 and X3 are independently NH or S; Y1 and Y2 are independently [ka] and Z1, Z2, Z3, and Z4 are each independently C, N, O, or S; R5, R6, and R7 are independently hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 15 is -OH or -NR9R 10 and Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R 11 , R 12 , R 13 , and R 14 are independently absent, hydrogen, or C 1-4 is alkyl, n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, at least one of G and G1 is C-X1-R2, and R2 is [ka] and / or At least one of R1 and R3 is [ka] is.

[0014] In another aspect, the present disclosure provides a compound of formula IA: [ka] (IA) or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: X1 is CH2, O or S; G is CH, C-SCH3, C-OCH3, or N; R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is —OH or —NH2; Y1 and Y2 are independently [ka] and X5 is S, O, or NR7; R5, R6, and R7 are independently hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 15 is -OH or -NR9R 10 and R9 and R 10 is independently selected from hydrogen and C1-C6 alkyl; Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, at least one R2 is [ka] and / or At least one of R1 and R3 is [ka] is.

[0015] In another aspect, the present disclosure provides a compound of formula IA': [ka] or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: X1 is CH2, O or S; G is CH, C-SCH3, C-OCH3, or N; R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is —OH or —NH2; Y1 and Y2 are independently [ka] and X5 is S, O, or NR7; R5, R6, and R7 are independently hydrogen, —OH, C1-C6 alkyl, [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, R2 is [ka] and / or At least one of R1 and R3 is [ka] is.

[0016] In yet another aspect, the present disclosure provides a compound of formula IB: [ka] (IB) or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: R1 and R3 are independently [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered ring; R 14 is hydrogen or C 1-4 is alkyl, R5, R6, and R7 are independently hydrogen, —OH, C1-C6 alkyl, [ka] is selected from w is an integer of 1 to 6, provided that at least one of R1 and R3 is [ka] is.

[0017] The compounds of the present application have been shown to be capable of activating STING protein function. Thus, the compounds of the present application are useful for treating diseases, disorders, or conditions treatable by activation of STING. Accordingly, the present application also includes a method for treating a disease, disorder, or condition treatable by activation of STING, comprising administering a therapeutically effective amount of one or more compounds of the present application to a subject in need thereof.

[0018] In a further embodiment, the compounds of the present application are used as pharmaceuticals. Thus, the present application also includes compounds of the present application for use as pharmaceuticals.

[0019] The present application also includes the use of one or more compounds of the present application for treating a disease, disorder, or condition treatable by activation of STING, as well as the use of one or more compounds of the present application for the preparation of a medicament for treating a disease, disorder, or condition treatable by activation of STING. The application further includes one or more compounds of the present application for use in treating a disease, disorder, or condition treatable by activation of STING.

[0020] The compounds of the present application are useful for treating diseases, disorders, or conditions mediated by STING protein activation. Accordingly, the present application also includes a method of treating a disease, disorder, or condition mediated by STING protein activation, comprising administering a therapeutically effective amount of one or more compounds of the present application to a subject in need thereof.

[0021] The present application also includes the use of one or more compounds of the present application for treating a disease, disorder, or condition mediated by STING protein activation, as well as the use of one or more compounds of the present application for the preparation of a medicament for treating a disease, disorder, or condition mediated by STING protein activation. The application further includes one or more compounds of the present application for use in the treatment of a disease, disorder, or condition mediated by STING protein activation.

[0022] In one embodiment, the disease, disorder, or condition mediated by STING protein activation or treatable by activation of STING is a neoplastic disorder. In one embodiment, treatment comprises the administration or use of one or a compound of the present application in an amount effective to ameliorate at least one symptom of the neoplastic disorder, e.g., reduce cell proliferation or reduce tumor burden, in a subject in need of such treatment.

[0023] In one embodiment, the disease, disorder, or condition mediated by STING protein activation or treatable by activation of STING is cancer.

[0024] In one embodiment, the disease, disorder, or condition mediated by STING protein activation or treatable by activation of STING is a disease, disorder, or condition associated with unregulated and / or abnormal cellular activity that is affected directly or indirectly by STING, hi another embodiment, the unregulated and / or abnormal cellular activity that is affected directly or indirectly by STING is proliferative activity in cells.

[0025] The present application also includes a method for inhibiting proliferative activity in a cell, comprising administering to the cell an effective amount of one or more compounds of the present application.

[0026] In a further embodiment, the disease, disorder, or condition mediated by STING protein activation or treatable by activation of STING is cancer, and one or more compounds of the present application are administered in combination with one or more additional cancer treatments, hi another embodiment, the additional cancer treatment is selected from radiation therapy, chemotherapy, targeted therapy, such as antibody therapy and small molecule therapy, including other tyrosine kinase inhibitors, immunotherapy, hormonal therapy, and anti-angiogenic therapy.

[0027] The present application further provides processes for the preparation of compounds of Formula I. General and specific methods are discussed and described in more detail in the Examples below.

[0028] Other features and advantages of the present application will become apparent from the following detailed description. It should be understood, however, that the detailed description and specific examples, while indicating embodiments of the present application, are given by way of illustration only, since various changes and modifications within the spirit and scope of the present application will become apparent to those skilled in the art from this detailed description. [Brief explanation of the drawings]

[0029] [Figure 1A] ~ [Figure 1B] FIG. 1 shows THP-1 plots for exemplary compounds IA and ID of the present disclosure. [Figure 2A] ~ [Figure 2D] FIG. 1 shows the STING haplotype curves of exemplary compounds IA and ID of the present disclosure. [Figure 3A] ~ [Figure 3B] FIG. 1 shows representative PBMC curves for exemplary compounds IA and ID of the present disclosure. [Figure 4A] ~ [Figure 4B] FIG. 1 shows additional representative PBMC curves for exemplary compounds IA and ID of the present disclosure. [Figure 5A] ~ [Figure 5B] FIG. 1 shows additional representative PBMC curves for exemplary compounds IA and ID of the present disclosure. [Figure 6A] ~ [Figure 6E] FIG. 1 shows tumor growth data measured using BioVolume after administration of various doses of exemplary Compound ID or vehicle control. [Figure 7A] FIG. 1 shows tumor IFNα levels after administration of various doses of an exemplary compound ID or vehicle control. [Figure 7B] FIG. 1 shows tumor IFNα levels after administration of various doses of the comparative compound diABZI or vehicle control. [Figure 8A] FIG. 1 shows tumor IFNγ levels after administration of various doses of an exemplary compound ID or vehicle control. [Figure 8B] FIG. 1 shows tumor IFNγ levels after administration of various doses of the comparative compound diABZI or vehicle control. [Figure 9A] ~ [Figure 9E] FIG. 1 shows data on tumor growth measured with calipers after administration of various doses of the comparative compound diABZI or vehicle control. [Figure 10] FIG. 1 shows the STING haplotype curves of exemplary compounds XV and XX of the present disclosure. [Figure 11]FIG. 1 shows THP-1 plots of exemplary compounds XV and XX of the present disclosure. [Figure 12-1] ~ [Figure 12-4] FIG. 1 shows representative PBMC curves for exemplary compounds XV and XX of the present disclosure. [Figure 13-1] ~ [Figure 13-3] FIG. 1 shows the activity of exemplary compounds XXVIII-XXXV of the present disclosure. [Figure 14] FIG. 1 shows THP-1 plots of exemplary compounds XXVIII-XXXV of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0030] definition Unless otherwise defined, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed embodiments belong.

[0031] As used herein, the terms "a" or "an" mean "at least one" or "one or more," unless the context clearly indicates otherwise.

[0032] As used herein, the term "about" means that the recited numerical value is an approximation and that small variations do not significantly affect the practice of the disclosed embodiments. When numerical values ​​are used, unless the context dictates otherwise, "about" means that the numerical value can vary ±10% and remain within the range of the disclosed embodiments.

[0033] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0034] As used herein, the term "alkenyl" refers to a straight- or branched-chain alkyl group having 2 to 20 carbon atoms and having one or more carbon-carbon double bonds. In some embodiments, alkenyl groups have 2 to 10 carbon atoms, 2 to 8 carbon atoms, 2 to 6 carbon atoms, 2 to 4 carbon atoms, 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 or 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-methyl-1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, and the like.

[0035] As used herein, the term "alkoxy" refers to a straight- or branched-chain -O-alkyl group having 1 to 20 carbon atoms. In some embodiments, the alkoxy group has 1 to 10 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, 2 to 10 carbon atoms, 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, t-butoxy, and the like.

[0036] As used herein, the term "alkyl" refers to a saturated hydrocarbon group that is straight-chained or branched. In some embodiments, an alkyl group has 1 to 20 carbon atoms, 2 to 20 carbon atoms, 1 to 10 carbon atoms, 2 to 10 carbon atoms, 1 to 8 carbon atoms, 2 to 8 carbon atoms, 1 to 6 carbon atoms, 2 to 6 carbon atoms, 1 to 4 carbon atoms, 2 to 4 carbon atoms, 1 to 3 carbon atoms, or 2 or 3 carbon atoms. Examples of alkyl groups include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, t-butyl, isobutyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), hexyl, isohexyl, heptyl, octyl, nonyl, 4,4-dimethylpentyl, 2,2,4-trimethylpentyl, decyl, undecyl, dodecyl, 2-methyl-1-propyl ... 2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2-methyl-1-pentyl, 2,2-dimethyl-1-propyl, 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, and the like.

[0037] As used herein, the term "alkylamino" refers to an amino group substituted with an alkyl group. In some embodiments, the alkyl group is a lower alkyl group having 1 to 6 carbon atoms. Alkylamino groups include, but are not limited to, -NHCH2CH3, -NH(CH2)2CH3, -NH(CH2)3CH3, -NH(CH2)4CH3, and -NH(CH2)5CH3.

[0038] As used herein, the term "alkylene" or "alkylenyl" refers to a divalent alkyl linking group. Examples of alkylene (or alkylenyl) include, but are not limited to, methylene or methyleneyl (-CH-), ethylene or ethylenyl (-CH-CH-), and propylene or propyleneyl (-CH-CH-CH-).

[0039] As used herein, the term "alkylthio" refers to an -S-alkyl group having 1 to 6 carbon atoms. Examples of alkylthio groups include, but are not limited to, -SCH2CH3, -S(CH2)2CH3, -S(CH2)3CH3, -S(CH2)4CH3, and -S(CH2)5CH3.

[0040] As used herein, the term "alkynyl" refers to a straight- or branched-chain alkyl group having 2 to 20 carbon atoms and one or more carbon-carbon triple bonds. In some embodiments, alkynyl groups have 2 to 10 carbon atoms, 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Examples of alkynyl groups include, but are not limited to, acetylene, 1-propylene, 2-propylene, and the like.

[0041] As used herein, the term "amino" means --NH.sub.2.

[0042] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group. Examples of aminoalkyl groups include, but are not limited to, -CHNH, -CHCHNH, -(CH)NH, -(CH)NH, and the like.

[0043] As used herein, the term "aminosulfonyl" means -S(=O)2NH2.

[0044] As used herein, the term "aryl" refers to a monocyclic, bicyclic, or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon. In some embodiments, aryl groups have 6 to 20 carbon atoms or 6 to 10 carbon atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, and tetrahydronaphthyl.

[0045] As used herein, the term "arylene" means an aryl linking group, ie, an aryl group that links one group to another group in a molecule.

[0046] As used herein, the term "carbamoyl" means -C(=O)-NH2.

[0047] As used herein, the term "carbocycle" means a 5- or 6-membered saturated or unsaturated cyclic ring that may contain an O, S, or N atom as part of the ring. Examples of carbocycles include, but are not limited to, cyclopentyl, cyclohexyl, cyclopenta-1,3-diene, phenyl, and any of the heterocycles listed herein.

[0048] As used herein, the term "carrier" refers to a diluent, adjuvant, or excipient with which the compound is administered in a composition.

[0049] As used herein, the term "compound" refers to all stereoisomers, tautomers, isotopes, and polymorphs of the compounds described herein.

[0050] As used herein, the terms "comprising" (and any form of comprising, such as "comprise," "comprises," and "comprised"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "includes" and "include"), or "containing" (and any form of containing, such as "contains" and "contain") are inclusive and open-ended, and include the options that follow the term and do not exclude additional, unrecited elements or method steps.

[0051] As used herein, the term "contacting" means bringing two compounds, molecules, or entities together in an in vitro system or an in vivo system.

[0052] As used herein, the term "cycloalkyl" refers to a non-aromatic cyclic hydrocarbon, including cyclized alkyl, alkenyl, and alkynyl groups, having up to 20 ring-forming carbon atoms. Cycloalkyl groups have 3 to 15 ring-forming carbon atoms, 3 to 10 ring-forming carbon atoms, 3 to 8 ring-forming carbon atoms, 3 to 6 ring-forming carbon atoms, 4 to 6 ring-forming carbon atoms, 3 to 5 ring-forming carbon atoms, or 5 or 6 ring-forming carbon atoms. Ring-forming carbon atoms of a cycloalkyl group may be optionally substituted with oxo or sulfido. Cycloalkyl groups include monocyclic and polycyclic ring systems, including, but not limited to, fused, bridged, and spirocyclic ring systems. In some embodiments, polycyclic ring systems contain 2, 3, or 4 fused rings. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, adamantyl, etc. Cycloalkyl groups can also have one or more aromatic rings fused (having a common bond) to the cycloalkyl ring, such as, for example, benzo or thienyl derivatives of pentane, pentene, hexane, etc. (e.g., 2,3-dihydro-1H-inden-1-yl, or 1H-inden-2(3H)-on-1-yl).

[0053] As used herein, the term "cycloalkylalkyl" refers to a C substituted with cycloalkyl. 1-6 It means alkyl.

[0054] As used herein, the term "dialkylamino" refers to an amino group substituted with two alkyl groups. In some embodiments, one or both of the alkyl groups have 1 to 6 carbon atoms.

[0055] As used herein, the term "heteroaryl" refers to an aromatic heterocycle having up to 20 ring-forming atoms (e.g., C) and at least one heteroatom ring member (ring atom) such as sulfur, oxygen, or nitrogen. In some embodiments, a heteroaryl group has at least one or more heteroatom ring-forming atoms, each of which is independently sulfur, oxygen, or nitrogen. In some embodiments, a heteroaryl group has 3 to 20 ring-forming atoms, 3 to 10 ring-forming atoms, 3 to 6 ring-forming atoms, or 3 to 5 ring-forming atoms. In some embodiments, a heteroaryl group contains 2 to 14 carbon atoms, 2 to 7 carbon atoms, or 5 or 6 carbon atoms. In some embodiments, a heteroaryl group has 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 or 2 heteroatoms. Heteroaryl groups include monocyclic and polycyclic (e.g., having 2, 3, or 4 fused rings) systems.Examples of heteroaryl groups include pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyridinyl (including 2-aminopyridine), triazinyl, furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl (such as indol-3-yl), pyrryl, oxazolyl, benzofuryl, benzothienyl, pyrazolyl, benzothiazolyl, isoxazolyl, triazolyl (including 1,2,4-triazole, 1,2,3-triazole, and 5-amino-1,2,4-triazole), tetrazolyl, indazolyl, isothiazolyl, 1,2,4-thiadiazolyl, benzothienyl, purinyl, carbazolyl, isoxazolyl, benzimidazolyl, indolinyl, and pyranyl. , pyrazolyl, triazolyl, oxadiazolyl (including 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 3-amino-1,2,4-oxadiazole, 1,3,4-oxadiazole), thianthrenyl, indolizinyl, isoindolyl, isobenzofuranyl, pyrrolyl, benzoxazolyl, xanthenyl, 2H-pyrrolyl, 3H-indolyl, 4H-quinolizinyl, phthalazinyl, acridinyl, naphthyridinyl, quinazolinyl, phenanthridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl, phenoxazinyl groups and the like.

[0056] As used herein, the term "heteroarylalkyl" refers to a C substituted with a heteroaryl group. 1-6 It means an alkyl group.

[0057] As used herein, the term "heteroarylamino" refers to an amino group substituted with a heteroaryl group.

[0058] As used herein, the term "heteroarylene" means a heteroaryl linking group, ie, a heteroaryl group that links one group to another group in a molecule.

[0059] As used herein, the term "heterocycle" or "heterocyclic ring" refers to a 5- to 7-membered monocyclic or 7- to 10-membered bicyclic ring system, any ring of which may be saturated or unsaturated, consisting of carbon atoms and 1-3 heteroatoms selected from N, O, and S, where the N and S heteroatoms may be optionally oxidized and the N heteroatom may be optionally quaternized, including any bicyclic group in which any of the heterocyclic rings defined above are fused to a benzene ring. Heterocycles include rings containing one oxygen or sulfur, one to three nitrogen atoms, or one oxygen or sulfur in combination with one or two nitrogen atoms. The heterocyclic ring may be attached at any heteroatom or carbon atom that results in the creation of a stable structure. Examples of heterocyclic groups include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, pyridyl, imidazolidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolin ... These include, but are not limited to, azolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, oxadiazolyl, and the like.

[0060] As used herein, the term "heterocycloalkyl" refers to a non-aromatic heterocycle having up to 20 ring-forming atoms, including cyclized alkyl, alkenyl, and alkynyl groups, in which one or more of the ring-forming carbon atoms are replaced with a heteroatom, such as an O, N, or S atom. Heterocycloalkyl groups can be monocyclic or polycyclic (e.g., fused, bridged, or spiro). In some embodiments, heterocycloalkyl groups have 1 to 20 carbon atoms or 3 to 20 carbon atoms. In some embodiments, heterocycloalkyl groups contain 3 to 14 ring-forming atoms, 3 to 7 ring-forming atoms, or 5 or 6 ring-forming atoms. In some embodiments, heterocycloalkyl groups have 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 or 2 heteroatoms. In some embodiments, heterocycloalkyl groups have 0 to 3 double bonds. In some embodiments, heterocycloalkyl groups have 0 to 2 triple bonds. Examples of heterocycloalkyl groups include, but are not limited to, morpholino, piperazinyl, thiomorpholino, tetrahydrofuranyl, tetrahydrothienyl, 2,3-dihydrobenzofuryl, piperidinyl, 1,3-benzodioxole, benzo-1,4-dioxane, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, pyrazolidinyl, thiazolidinyl, imidazolidinyl, pyrrolidin-2-one-3-yl, etc. Furthermore, ring-forming carbon atoms and heteroatoms of heterocycloalkyl groups can be substituted with oxo or sulfido. For example, ring-forming S atoms can be substituted with one or two oxo atoms (forming S(O) or S(O)2). As another example, ring-forming C atoms can be substituted with oxo (forming carbonyl).Heterocycloalkyl groups can also have one or more aromatic rings fused (having a common bond) to a non-aromatic heterocyclic ring, including, but not limited to, pyridinyl, thiophenyl, phthalimidyl, naphthalimidyl, and benzo derivatives of heterocycles, such as indolene, isoindolene, 5,6-dihydrothieno[2,3-c]pyridin-7(4H)-one-5-yl, isoindolin-1-one-3-yl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridin-5-yl, and 3,4-dihydroisoquinolin-1(2H)-one-3-yl groups. The ring-forming carbon atoms and heteroatoms of heterocycloalkyl groups can be optionally substituted with oxo or sulfido.

[0061] As used herein, the term "heterocycloalkylalkyl" refers to an alkyl group substituted with a heterocycloalkyl. In some embodiments, the alkyl group is C 1-6 It is an alkyl group.

[0062] As used herein, the term "hydroxy" or "hydroxyl" refers to an --OH group.

[0063] As used herein, the term "hydroxyalkyl" or "hydroxylalkyl" refers to an alkyl group substituted with a hydroxyl group. Examples of hydroxylalkyl groups include, but are not limited to, -CHOH and -CHCHOH.

[0064] As used herein, the terms "individual," "subject," and "patient," used interchangeably, refer to any animal described herein.

[0065] As used herein, the phrase "in need thereof" means that an "individual," "subject," or "patient" has been identified as having need of a particular method, prevention, or treatment. In some embodiments, identification may be by any diagnostic means. An "individual," "subject," or "patient" may be in need of any of the methods, preventions, and treatments described herein. In some embodiments, an "individual," "subject," or "patient" is in, moves to, or has moved to an environment where a particular disease, disorder, or condition is prevalent.

[0066] As used herein, the term "integer" refers to a natural number. For example, "an integer from 1 to 5" means 1, 2, 3, 4, or 5.

[0067] All cyclic groups contain one or more rings (i.e., are polycyclic). When a cyclic group contains more than one ring, the rings may be fused, bridged, or joined by bonds.

[0068] A first ring is "fused" to a second ring means that the first ring and the second ring share two adjacent atoms between them.

[0069] As used herein, the term "isolated" means that a compound described herein, or a pharmaceutically acceptable salt thereof, is separated, for example by conventional techniques, from other components of either a) a natural source such as a plant or cell, e.g., a bacterial culture, or b) a synthetic organic chemical reaction mixture.

[0070] As used herein, the term "n-membered," where n is an integer, typically refers to the number of ring-forming atoms in a moiety, where n is the number of ring-forming atoms. For example, pyridine is an example of a 6-membered heteroaryl ring, and thiophene is an example of a 5-membered heteroaryl ring.

[0071] As used herein, the phrase "optionally substituted" means that substitution is optional and thus includes both unsubstituted and substituted atoms and moieties. A "substituted" atom or moiety indicates that any hydrogen atom on the specified compound or moiety can be replaced with a group selected from the listed substituents, provided that the normal valence of the specified compound or moiety is not exceeded and the substitution results in a stable compound. For example, if a methyl group is optionally substituted, one, two, or three hydrogen atoms on a carbon atom in the methyl group can be replaced with one, two, or three of the listed substituents.

[0072] As used herein, the phrase "pharmaceutically acceptable" means that a compound, material, composition, and / or dosage form is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other animals. In some embodiments, "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia for use in animals, more particularly humans. In some embodiments, a pharmaceutically acceptable compound, material, composition, and / or dosage form produces no lasting adverse effects on the subject or on the overall health of the treated subject. However, it will be recognized that transient effects, such as a mild irritation or a "tingling" sensation, are common with the administration of pharmaceuticals, and the presence of such transient effects is not inconsistent with the composition, formulation, or ingredients (e.g., excipients) in question.

[0073] As used herein, the phrase "pharmaceutically acceptable salts" includes, but is not limited to, salts of acidic or basic groups. Compounds that are basic in nature can form a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds include non-toxic acid addition salts, i.e., sulfuric acid, thiosulfuric acid, citric acid, maleic acid, acetic acid, oxalic acid, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, hydrogen sulfide, bisulfite, phosphate, acid phosphate, isonicotinate, borate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, glutamic acid ... They form salts containing pharmacologically acceptable anions, including, but not limited to, conate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, bicarbonate, malonate, mesylate, esylate, napsydisylate, tosylate, besylate, orthophosphate, trifluoroacetate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid)) salts. Compounds containing an amino moiety can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. Compounds that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include, but are not limited to, alkali metal or alkaline earth metal salts, particularly calcium, magnesium, ammonium, sodium, lithium, zinc, potassium, and iron salts. Salts also include quaternary ammonium salts of the compounds described herein, which compounds have one or more tertiary amine moieties.

[0074] As used herein, the term "phenyl" means -C6H5. A phenyl group can be unsubstituted or substituted with one, two, or three suitable substituents.

[0075] As used herein, the terms "prevention" or "preventing" refer to a reduction in the risk of acquiring a particular disease, condition, or disorder.

[0076] As used herein, the phrase "solubilizing agent" means an agent that results in the formation of a micellar or true solution of a drug.

[0077] As used herein, the term "solution / suspension" refers to a liquid composition in which a first portion of the active agent is in solution and a second portion of the active agent is in particulate form in suspension in a liquid matrix.

[0078] As used herein, the phrase "suitable substituent" or "substituent" refers to a group that does not abrogate the synthetic or pharmaceutical utility of the compounds described herein or intermediates useful in preparing them. Examples of suitable substituents include, but are not limited to, C-C alkyl, C-C alkenyl, C-C alkynyl, C-C aryl, C-C alkoxy, C-C heteroaryl, C-C cycloalkyl, C-C aryloxy, -CN, -OH, oxo, halo, haloalkyl, -NO, -COH, -NH, -NH(C-C alkyl), -N(C-C alkyl), -NH(C aryl), -N(C-C aryl), -CHO, -CO(C-C alkyl), -CO((C-C) aryl), -CO((C-C) alkyl), and -CO((C-C) aryl). One of ordinary skill in the art can readily select appropriate substituents based on the stability and pharmacological and synthetic activity of the compounds described herein.

[0079] In embodiments of the present application, the compounds described herein may have at least one asymmetric center. When a compound has more than one asymmetric center, the asymmetric center may exist as a diastereomer. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present application. While the stereochemistry of a compound may be as shown in any given compound listed herein, it is further understood that such compounds may also contain a certain amount (e.g., less than 20%, suitably less than 10%, more suitably less than 5%) of compounds of the present application having an alternative stereochemistry. Any optical isomer, whether as an isolated, pure, or partially purified optical isomer, or racemic, or other mixture, is intended to be encompassed within the scope of the present application.

[0080] The compounds of the present application may exist in different tautomeric forms, and it is intended that any tautomeric form that the compounds of the present invention form is included within the scope of the present application.

[0081] The compounds of the present application may further exist in different polymorphic forms, and it is contemplated that any polymorphs formed are included within the scope of the present application.

[0082] As used herein, the phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or medical response sought in a tissue, system, animal, individual, or human by a researcher, veterinarian, physician, or other clinician. The therapeutic effect depends on the disorder being treated or the desired biological effect. Thus, the therapeutic effect may be a reduction in the severity of symptoms associated with the disorder and / or an inhibition (partially or completely) of the progression of the disorder, or an improvement in the treatment, cure, prevention, or elimination of the disorder, or an amelioration of side effects. The amount necessary to elicit a therapeutic response can be based, for example, on the age, health, size, and sex of the subject. The optimal amount can also be determined by monitoring the subject's response to treatment.

[0083] At various places in the present specification, substituents of compounds may be disclosed in groups or in ranges. It is specifically intended that the present disclosure include any and all individual subcombinations of the members of such groups and ranges. For example, "C 1-6 The term "alkyl" is specifically intended to individually disclose methyl, ethyl, propyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0084] For compounds in which a variable appears more than once, each variable may be a different moiety selected from a Markush group that provides options for the variable. For example, if a structure is described with two R groups occurring simultaneously on the same compound, the two R groups may represent different moieties selected from the Markush group defined for R. In another example, any of the multiple substituents "R" may be, for example, [ka] It is understood that when shown in the form: the substituent "R" can occur "x" times at any position on the ring, and "R" can be a different moiety at each occurrence. Further, in the above example, when the variable "Y" normally contains one or more hydrogens, for example, when "Y" is CH, NH, etc., any H can be replaced with a substituent.

[0085] The terms "treating" or "treatment," as used herein and as well understood in the art, refer to an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of disease, stabilized (i.e., not worsening) disease, prevention of disease spread, delay or slowing of disease progression, improvement or relief of disease state, reduction in disease recurrence, and remission (partial or total). "Treating" and "treatment" can also mean prolonging survival compared to expected survival in the absence of treatment. As used herein, "treating" and "treatment" also include prophylactic treatment. For example, a subject with early-stage cancer can be treated to prevent progression, or alternatively, a subject in remission can be treated with a compound or composition of the present application to prevent recurrence. A method of treatment includes administering a therapeutically effective amount of one or more compounds of the present application to a subject, and can consist of a single dose or, alternatively, can include a series of doses. For example, in some embodiments, the compound of the present application can be administered at least once a week. In some embodiments, the compound can be administered to a subject about once every three weeks, or about once a week to about once a day for a given treatment. In another embodiment, the compound is administered two, three, four, five, or six times a day. The length of the treatment period depends on various factors, such as the severity of the disease, disorder, or condition, the age of the subject, the concentration and / or activity of the compound of the present application, and / or a combination thereof. It will also be understood that the effective dosage of the compound used for treatment may increase or decrease over the course of a particular treatment regimen. Changes in dosage may occur and be evident using standard diagnostic assays known in the art. In some cases, chronic administration may be required. For example, the compound is administered to a subject in an amount and for a period sufficient to treat the patient.

[0086] "Alleviating" a disease or disorder means that the severity and / or undesirable clinical symptoms of the disorder or disease state are reduced and / or the time course of progression is slowed or prolonged compared to if the disorder is not treated.

[0087] As used herein, the terms "prevention" or "prophylaxis," or synonyms thereof, refer to a reduction in the risk or probability that a patient will suffer from or develop symptoms associated with a disease, disorder, or condition that is mediated by or treatable by STING protein activation.

[0088] As used herein, a "STING-mediated disease, disorder, or condition" refers to a disease, disorder, or condition that is treatable by activation of STING activity, particularly using a STING agonist such as one or more of the compounds of the present application described herein.

[0089] As used herein, the term "mediated by STING" means that the disease, disorder, or condition being treated is affected, modulated, and / or has, either directly or indirectly, some biological basis that involves aberrant STING activity, particularly reduced STING activity resulting from mutations, splice variations, etc. These diseases respond favorably when disease-associated STING activity is activated by one or more of the compounds of the present application.

[0090] As used herein, the term "STING agonist" refers to a compound that is an "activator of STING" (i.e., activates STING activity), and therefore the terms "activator" and "agonist" may be used interchangeably with respect to the compounds of the present application.

[0091] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of one or more compounds of the present application effective, at dosages and for periods of time necessary, to achieve a desired result. For example, in the context of treating a disease, disorder, or condition mediated by STING protein activation or treatable by STING activation, an effective amount is, for example, an amount that increases STING protein activation or increases STING activity compared to the activity in the absence of the one or more compounds. An effective amount may vary depending on factors such as the disease state, age, sex, and / or weight of the subject. The amount of a given compound corresponding to such an amount will vary depending on various factors, such as the given drug or compound, pharmaceutical formulation, route of administration, type of condition, disease, or disorder, and characteristics of the subject being treated, but can nevertheless be routinely determined by one of skill in the art. In one embodiment, an effective amount is an amount that results in an improvement or reduction of any disease symptoms following treatment therewith. When the disease is cancer, an effective amount may, for example, cause a reduction in the number, growth rate, size, and / or distribution of tumors.

[0092] As used herein, the term "administered" means administering a therapeutically effective amount of one or more compounds or compositions of the present application to a cell, either in cell culture or in a subject.

[0093] As used herein, the term "neoplastic disorder" refers to a disease, disorder, or condition characterized by cells capable of autonomous growth or replication, e.g., an abnormal state or condition characterized by proliferative cell growth. As used herein, the term "neoplasm" refers to a mass of tissue resulting from the abnormal growth and / or division of cells in a subject with a neoplastic disorder. Neoplasms may be benign (e.g., uterine fibroids and pigmented nevi), potentially malignant (e.g., carcinoma in situ), or malignant (i.e., cancer). Exemplary neoplastic disorders include, but are not limited to, carcinomas, sarcomas, metastatic disorders (e.g., tumors arising from the prostate), hematopoietic neoplastic disorders (e.g., leukemia, lymphoma, myeloma, and other malignant plasma cell disorders), metastatic tumors, and other cancers. Common cancers include breast cancer, prostate cancer, colon cancer, lung cancer, liver cancer, brain tumors, ovarian cancer, and pancreatic cancer.

[0094] As used herein, the term "cancer" refers to a cell proliferative disease state, including acute lymphoblastic leukemia, adult; acute lymphoblastic leukemia, childhood; acute myeloid leukemia, adult; adrenocortical carcinoma; adrenocortical carcinoma, childhood; AIDS-related lymphoma; AIDS-related malignancy; anal cancer; astrocytoma, childhood cerebellar; astrocytoma, childhood cerebral; bile duct cancer, extrahepatic; bladder cancer; bladder cancer, childhood; bone cancer, osteosarcoma / malignant fibrous histiocytoma; brain stem glioma, childhood; brain tumor, adult; brain tumor, brain stem glioma, childhood; brain tumor, cerebellar astrocytoma, childhood; brain tumor, cerebral astrocytoma / malignant fibrous histiocytoma Glioma, childhood;Brain tumor, ependymoma, childhood;Brain tumor, medulloblastoma, childhood;Brain tumor, supratentorial primitive neuroectodermal tumor, childhood;Brain tumor, visual pathway and hypothalamic glioma, childhood;Brain tumor, childhood (other);Breast cancer;Breast cancer and pregnancy;Breast cancer, childhood;Breast cancer, male;Bronchial adenoma / carcinoid, childhood;Carcinoid tumor, childhood;Carcinoid tumor, gastrointestinal;Carcinoma, adrenal cortex;Carcinoma, pancreatic islet cell;Carcinoma of unknown primary;Central nervous system lymphoma, primary;Cerebellar astrocytoma, childhood;Cerebral astrocytoma / malignant glioma, childhood;Cervical cancer;Childhood cancer;Chronic lymphocytic leukemia;Chronic Myeloid leukemia;Chronic myeloproliferative disorders;Clear cell sarcoma tendon sheath;Colon cancer;Colorectal cancer, childhood;Cutaneous T-cell lymphoma;Uterine cancer;Ependymoma, childhood;Epithelial carcinoma, ovarian;Esophageal cancer;Esophageal cancer, childhood;Ewing family tumors;Extracranial germ cell tumors, childhood;Extragonadal germ cell tumors;Extrahepatic bile duct cancer;Eye cancer, intraocular melanoma, eye cancer, retinoblastoma;Gallbladder cancer;Gastric (stomach) cancer;Gastric (stomach) cancer, childhood;Gastrointestinal carcinoid tumors;Extracranial germ cell tumors, childhood;Extragonadal germ cell tumors;Germ cell Alveolar tumor, ovarian;Gestational trophoblastic tumor;Glioma, childhood brainstem;Glioma, childhood visual pathway and hypothalamic;Hairy cell leukemia;Head and neck cancer;Hepatocellular (liver) cancer, adult (primary);Hepatocellular (liver) cancer, childhood (primary);Hodgkin's lymphoma, adult;Hodgkin's lymphoma, childhood;Hodgkin's lymphoma during pregnancy;Hypopharyngeal cancer;Hypothalamic and visual pathway glioma, childhood;Intraocular melanoma;Islet cell carcinoma (pancreatic islet);Kaposi's sarcoma;Kidney cancer;Laryngeal cancer;Laryngeal cancer, childhood;Leukemia, acute lymphoblastic, adult;Leukemia, acute lymphoblastic, childhood;Leukemia, acute myeloid, adult;Leukemia, acute myeloid, childhood; Leukemia, chronic lymphocytic; Leukemia, chronic myeloid; Leukemia, hairy cell; Lip and oral cancer; Liver cancer, adult (primary); Liver cancer, childhood (primary); Lung cancer, non-small cell; Lung cancer, small cell; Lymphoblastic leukemia, adult acute; Lymphoblastic leukemia, childhood (acute); Lymphocytic leukemia, chronic; Lymphoma, AIDS-related; Lymphoma, central nervous system (primary); Lymphoma, cutaneous T-cell; Lymphoma, Hodgkin, adult; Lymphoma, Hodgkin, childhood; Lymphoma, Hodgkin during pregnancy; Lymphoma, non-Hodgkin, adult; Lymphoma, non-Hodgkin, childhood ;Lymphoma, Pregnancy, Non-Hodgkin's;Lymphoma, Primary Central Nervous System;Macroglobulinemia, Waldenstrom;Male Breast Cancer;Malignant Mesothelioma, Adult;Malignant Mesothelioma, Childhood;Malignant Thymoma;Medulloblastoma, Childhood;Melanoma;Melanoma, Intraocular;Merkel Cell Carcinoma;Mesothelioma, Malignant;Metastatic Squamous Neck Cancer of Unknown Primary;Multiple Endocrine Neoplasia Syndrome, Childhood;Multiple Myeloma / Plasma Cell Neoplasm;Mycosis Fungoides;Myelodysplastic Syndrome;Myeloid Leukemia, Chronic;Myeloid Leukemia, Acute Childhood;Myeloma, Multiple;Myeloproliferative Disorders, Chronic;Nasal Cavity and Paranasal Sinus Cancer;Nasopharyngeal Carcinoma;Nasopharyngeal Carcinoma, Childhood;Neurologic Blastomas; Non-Hodgkin's lymphoma, adult; Non-Hodgkin's lymphoma, childhood; Non-Hodgkin's lymphoma during pregnancy; Non-small cell lung cancer; Oral cavity cancer, childhood; Oral cavity and lip cancer; Oropharyngeal cancer; Osteosarcoma / malignant fibrous histiocytoma of bone; Ovarian cancer, childhood; Ovarian epithelial cancer; Ovarian germ cell tumor; Ovarian low malignant potential tumor; Pancreatic cancer; Pancreatic cancer, childhood; Pancreatic islet cell cancer; Cancer of the paranasal sinuses and nasal cavity; Parathyroid cancer; Penile cancer; Pheochromocytoma; Pineal and supratentorial primitive neuroectodermal tumors, childhood; Pituitary tumors; Plasma cell neoplasms / multiple myeloma; Pleuropulmonary blastoma; Pregnancy and breast cancer; Pregnancy and Hodgkin's lymphoma ;Pregnancy and non-Hodgkin's lymphoma;Primary central nervous system lymphoma;Primary liver cancer, adult;Primary liver cancer, childhood;Prostate cancer;Rectal cancer;Renal cell (kidney) cancer;Renal cell carcinoma, childhood;Renal pelvis and ureter, transitional cell carcinoma;Retinoblastoma;Rhabdomyosarcoma, childhood;Salivary gland cancer;Salivary gland cancer, childhood;Sarcoma, Ewing family tumor;Sarcoma, Kaposi's tumor;Sarcoma (osteosarcoma) / malignant fibrous histiocytoma of bone;Sarcoma, rhabdomyosarcoma, childhood;Sarcoma, soft tissue, adult;Sarcoma, soft tissue, childhood;Sezary syndrome;Skin cancer;Skin cancer, childhood;Skin cancer (melanoma);Skin carcinoma, Merkel cell;Cancers that may be treated include, but are not limited to, small cell lung cancer; small intestine cancer; soft tissue sarcoma, adult; soft tissue sarcoma, childhood; squamous cell carcinoma of the neck of unknown primary site, metastatic; gastric cancer; gastric cancer, childhood; supratentorial primitive neuroectodermal tumor, childhood; T-cell lymphoma, skin; testicular cancer; thymoma, childhood; thymoma, malignant; thyroid cancer; thyroid cancer, childhood; transitional cell carcinoma of the renal pelvis and ureter; trophoblastic tumor, gestational; carcinoma of unknown primary site, childhood; atypical carcinoma of childhood; transitional cell carcinoma of the ureter and renal pelvis; urethral cancer; uterine sarcoma; vaginal cancer; visual pathway and hypothalamic glioma, childhood; vulvar cancer; Waldenstrom's macroglobulinemia; and Wilms' tumor. Metastases of the aforementioned cancers can also be treated according to the methods described herein.

[0095] symbol [ka] is used herein to represent the point of attachment of a group to the rest of a molecule or chemical formula.

[0096] Suitable compounds described herein may also include tautomeric forms. Tautomeric forms result from the interchange of a single bond with an adjacent double bond, with the concomitant migration of a proton. Tautomeric forms include proton tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of proton tautomers include, but are not limited to, ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in a heterocyclic ring system, including, but not limited to, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms may be in equilibrium or sterically locked into one form by appropriate substitution.

[0097] The compounds described herein also include hydrated and solvated forms, as well as anhydrous and unsolvated forms.

[0098] In some embodiments, the compound or salt thereof is substantially isolated. Partial isolation can include, for example, a composition enriched in any one or more of the compounds described herein. Substantial isolation can include a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of any one or more of the compounds described herein or salts thereof. Methods for isolating compounds and their salts are routine in the art.

[0099] STING agonist compounds Compounds of the present application have been prepared and found to inhibit uncontrolled and / or abnormal cellular activity that is affected directly or indirectly by activation of the STING protein. In particular, the compounds of the present application exhibit activity as STING agonists and are therefore useful in therapy, for example, for the treatment of neoplastic disorders such as cancer.

[0100] It will be understood by those skilled in the art that the compounds of the present invention may exist in tautomeric forms, including but not limited to, Formula (A), Formula (B) and / or Formula (C), or zwitterionic forms, including but not limited to, Formula (D) or Formula (E): [ka] Formula (A) Formula (B) Formula (C) TIFF2026508759000031.tif32100 formula (D) formula (E)

[0101] Agonists of the stimulator of interferon genes (STING) represent a promising class of immunomodulators with potential for cancer treatment. This application relates to a novel class of STING agonists containing hydrazide and / or hydrazine functional groups.

[0102] Accordingly, the present application includes compounds of formula I or a solvate, pharmaceutically acceptable salt, or tautomer thereof: [ka] (I) During the ceremony, Ring A is [ka] (a) (b) (c) (d) is selected from the group consisting of During the ceremony, G and G1 are independently N, CH, or C-X1-R2; G' and G2 are independently N or CH; X is NR, O, or S; X' is N or CH; X1 is CH2, O or S; R is hydrogen or C 1-4 is alkyl, When G and G1 are each C-X1-R2, the R2 groups may be linked to form L1; L and L1 are each independently C 2-4 Alkylene or C 2-4 is alkenylene, R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is -OH or -NR9R 10 and R9 and R 10 is independently selected from hydrogen and C1-C6 alkyl; X2 and X3 are independently NH or S; Y1 and Y2 are independently [ka] and Z1, Z2, Z3, and Z4 are each independently C, N, O, or S; R5, R6, and R7 are independently hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 15 is -OH or -NR9R 10 and Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R 11 , R 12 , R 13 , and R 14 are independently absent, hydrogen, or C 1-4 is alkyl, n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, at least one of G and G1 is C-X1-R2, and R2 is [ka] and / or At least one of R1 and R3 is [ka] is.

[0103] In some embodiments, ring A is [ka] (a) (b) (c) (d) is selected from the group consisting of Structures (a), (b), (c), and (d) are attached to X2 and N of adjacent 5-membered heterocyclic rings in formula I to form an 8- or 9-membered aromatic heterofused ring system of formula I, wherein G and G1 are independently N, CH, or C-X1-R2, G' and G2 are independently N or CH, X is NR, O, or S, X' is N or CH, X1 is CH2, O, or S, and R is hydrogen or C 1-4 and X2 and X3 are each independently NH or S. In some embodiments, both X2 and X3 are N. In other embodiments, both X2 and X3 are NH. In other embodiments, X2 is NH and X3 is N. In still other embodiments, X2 is N and X3 is NH.

[0104] In Formula I, when ring A is structure (c), X is one carbon away from X on the adjacent fused 5-membered heterocyclic ring. When ring A is structure (d), X is one carbon away from the N atom on the adjacent fused 5-membered heterocyclic ring.

[0105] In some embodiments, Ring A is selected from phenyl, pyridinyl, imidazolyl, thiazolyl, thiophenyl, furanyl, and pyrrolyl, all of which are substituted with R3, wherein R3 is: [ka] R5, R6, and R7 are independently selected from the group consisting of hydrogen, C1-C6 alkyl, C2-C6 alkenyl, [ka] R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 15 is -OH or -NR9R 10 wherein ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; and R 11 , R 12 , R 13 , and R 14 are independently absent, hydrogen, or C 1-4 R and R in the amino group are alkyl. 10 is independently selected from hydrogen and C-C alkyl. In some embodiments, Ring C is selected from the group consisting of phenyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, pyrazolyl, oxathiolyl, isoxathiolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl.

[0106] In some embodiments, the linking group L is C 2-4 Alkylene or C 2-4 Preferably, L is ethylenyl, propylenyl, or butylenyl. In some embodiments, L is ethylenyl. In one embodiment, L is C2-alkenylene and X2 and X3 are NH.

[0107] In some embodiments, R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of:

[0108] Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocyclic ring containing 1-2 heteroatoms selected from N, O, and S. In some embodiments, Ring B is selected from the group consisting of phenyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, pyrazolyl, oxathiolyl, isoxathiolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Ring B is -OH or -NR9R 10 is substituted with R8,

[0109] In some embodiments, at least one of R5, R6, and R7 is a hydrogen atom.

[0110] In some embodiments, Y1 and Y2 are independently: [ka] Z1, Z2, Z3, and Z4 are each independently C, N, O, or S; R 11 , R 12 , R 13 , and R 14 are independently absent, hydrogen, or C 1-4 In some embodiments, Y and Y are independently selected from the group consisting of pyrrolyl, furanyl, thiophenyl, imidazolyl, pyrazolyl, oxathiolyl, isoxathiolyl, oxazolyl, isoxzolyl, thiazolyl, isothiazolyl, triazolyl, furazonyl, oxadiazolyl, thiadiazolyl, dioxazolyl, dithiazolyl, and tetrazolyl.

[0111] In some embodiments, R1 and R3 are independently: [ka] is selected from the group consisting of:

[0112] In some embodiments disclosed herein, at least one of G and G1 is C-X1-R2, wherein R2 is: [ka] and / or At least one of R1 and R3 is [ka] In some embodiments, both R and R are independently [ka] In other embodiments, both R1 and R3 are independently: [ka] The hydroxamic acid-containing hydrazide and hydrazine functional groups are compatible with different conjugation and linker chemistries, which can be used to generate a variety of STING-vector conjugates for developing tumor-targeted immunotherapies. Hydrazide- and hydrazine-derived linkers can be specifically cleaved within tumor cells and / or in the tumor microenvironment (e.g., extracellular matrix) by different cleavage mechanisms to release active STING agonist payloads. Such cleavage mechanisms include enzymatic cleavage by cathepsin B or legumain, chemical cleavage at low pH in lysosomes, and reductive cleavage by glutathione.

[0113] In the compounds of formula I, the variables n, p, q, t, and v are independently integers from 2 to 6, and the variables k, l, m, o, u, and w are independently integers from 1 to 6.

[0114] In some embodiments, the compound of Formula I is [ka] JPEG2026508759000051.jpg183170.

[0115] In another embodiment, the compound of formula I is [ka] JPEG2026508759000053.jpg169170.

[0116] Some embodiments disclosed herein include a subset of Formula I represented by Formula IA: [ka] (IA) or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: X1 is CH2, O or S; G is CH, C-SCH3, C-OCH3, or N; R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is —OH or —NH2; Y1 and Y2 are independently [ka] and X5 is S, O, or NR7; R5, R6, and R7 are independently hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 15 is -OH or -NR9R 10 and R9 and R 10 is independently selected from hydrogen and C1-C6 alkyl; Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, at least R2, [ka] and / or At least one of R1 and R3 is [ka] In some embodiments, both R and R are independently [ka] In other embodiments, both R1 and R3 are independently: [ka] is.

[0117] Some embodiments disclosed herein include a subset of Formula I represented by Formula IA': [ka] , or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: X1 is CH2, O or S; G is CH, C-SCH3, C-OCH3, or N; R2 is Hydrogen, C 2-4 Cyclic ethers, C 3-4 Cyclic alkyl, C 1-4 Alkyl, [ka] is selected from the group consisting of R1 and R3 are independently [ka] is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R8 is —OH or —NH2; Y1 and Y2 are independently [ka] and X5 is S, O, or NR7; R5, R6, and R7 are independently hydrogen, —OH, C1-C6 alkyl, [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, R2 is [ka] and / or At least one of R1 and R3 is [ka] is.

[0118] In some embodiments of formula IA', G is N, X1 is O, and R1 and R3 are each [ka] and R2 is [ka] wherein n is 2, 3, or 4; R5, R6, and R7 are each independently H, C1-C6 alkyl, and [ka] Y1 and Y2 are each selected from [ka] [wherein X5 is O].

[0119] In another embodiment of formula IA', G is N, X1 is O, and R1 and R3 are each [ka] and R2 is [ka] wherein n is 3, R5 is methyl, R6 and R7 are H, and Y1 and Y2 are each [ka] wherein X5 is O, so that the resulting structure is structure (XX): [ka] Derivatives and isomers of structure XX are those in which (with respect to formula IA') R5, R6 and R7 are each independently H, methyl or [ka] and a compound selected from the following structures XXI, XXII, XXIII, and XXIV: [ka] Includes.

[0120] In some embodiments, the compounds of formula IA and formula IA′ are [ka] JPEG2026508759000081.jpg197170, wherein R5, R6, R7, n, and m are as defined above.

[0121] In other embodiments, the compounds of formula IA and formula IA′ are [ka] wherein R5, R6, R7, n, and m are as defined above.

[0122] Some embodiments disclosed herein include a subset of Formula I represented by Formula IA': [ka] (IA'') or a solvate, pharmaceutically acceptable salt, or tautomer thereof, wherein: X1 is CH2, O or S; G is CH, C-OCH3, or N; R2 is [ka] and R3 is [ka] and Y1 and Y2 are independently [ka] and X5 is O or S; p is an integer of 2 to 6.

[0123] In some embodiments of formula IA'', R3 is [ka] G is C-OCH3, X1 is O, p is 3, and Y1 is [ka] is.

[0124] In other embodiments of formula IA'', R3 is [ka] G is C-OCH3, X1 is O, p is 3, and Y1 is [ka] and X5 is O.

[0125] In other embodiments of formula IA'', R3 is [ka] G is C-OCH3, X1 is O, p is 3, and Y1 is [ka] is.

[0126] In other embodiments of formula IA'', R3 is [ka] G is C-OCH3, X1 is O, p is 3, and Y1 is [ka] and X5 is O.

[0127] In still other embodiments of formula IA'', R3 is [ka] and G is N, X1 is O, p is 3, and Y1 is [ka] is.

[0128] In still other embodiments of formula IA'', R3 is [ka] and G is N, X1 is O, p is 3, and Y1 is [ka] and X5 is O.

[0129] In still other embodiments of formula IA'', R3 is [ka] and G is N, X1 is O, p is 3, and Y1 is [ka] is.

[0130] In still other embodiments of formula IA'', R3 is [ka] and G is N, X1 is O, p is 3, and Y1 is [ka] and X5 is O.

[0131] An embodiment of formula IA″ is the following compound: [ka] or a solvate thereof, a pharmaceutically acceptable salt thereof, or a tautomer thereof.

[0132] Ring A [ka] In some embodiments of Formula I, where G' and G2 are CH, G and G1 are each C-X1-R2, and X1 is CH2, O, or S, the second linking group L1 can be formed through the R2 group. Thus, in another embodiment, the subset of Formula I represented by Formula IB: [ka] (IB) or a solvate, pharmaceutically acceptable salt, or tautomer thereof is disclosed herein, wherein: R1 and R3 are independently [ka] wherein R5 and R6 may be joined to form a 5- or 6-membered heterocyclic ring; R 14 is hydrogen or C 1-4 is alkyl, R5, R6, and R7 are independently hydrogen, —OH, C1-C6 alkyl, [ka] is selected from w is an integer of 1 to 6, provided that at least one of R1 and R3 is [ka] In some embodiments, both R and R are independently [ka] is.

[0133] In another embodiment, the compound of formula IB is [ka] wherein R5, R6, and R7 are as defined above.

[0134] In some embodiments, the compound of the invention is selected from the compounds listed in Table 1. For any compound given a reference number, the reference number corresponds to the reference number in the Examples. [Table 1] JPEG2026508759000112.jpg215170 JPEG2026508759000113.jpg220170 JPEG2026508759000114.jpg235170 JPEG2026508759000115.jpg232170 JPEG2026508759000116.jpg224170 JPEG2026508759000117.jpg233170 JPEG2026508759000118.jpg226170 JPEG2026508759000119.jpg217170 JPEG2026508759000120.jpg245170 JPEG2026508759000121.jpg231170 JPEG2026508759000122.jpg148170

[0135] The compounds of the present application are suitably formulated into compositions using one or more carriers in a conventional manner. Accordingly, the present application also includes compositions comprising one or more compounds of the present application and a carrier. The compounds of the present application are suitably formulated into pharmaceutical compositions for administration to a subject in a biologically compatible form suitable for in vivo administration. Accordingly, the present application further includes pharmaceutical compositions comprising one or more compounds of the present application and a pharmaceutically acceptable carrier.

[0136] The compounds of the present application can be administered to a subject in various forms depending on the selected administration route, as will be understood by those skilled in the art. The compounds of the present application can be administered, for example, orally, parenterally, bucally, sublingually, nasally, rectally, by patch, pump, or transdermal administration, and the pharmaceutical composition will be formulated accordingly. Administration may also be by means of a pump for periodic or continuous delivery.

[0137] Parenteral administration includes intravenous, intraarterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, intranasal, intrapulmonary (e.g., by use of an aerosol), intrathecal, rectal, and topical (including use of a patch or other transdermal delivery device) modes of administration. Parenteral administration may also be by continuous infusion over a selected period of time. Conventional procedures and ingredients for the selection and preparation of appropriate compositions are known to those of skill in the art.

[0138] The compounds of the present application may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or may be enclosed in hard or soft shell gelatin capsules, or may be compressed into tablets, or may be directly incorporated with dietary food.For oral therapeutic administration, the compounds may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, caplets, pellets, granules, lozenges, chewing gum, powders, syrups, elixirs, wafers, aqueous solutions and suspensions, etc.For tablets, carriers used include lactose, corn starch, sodium citrate, and phosphate salts. Pharmaceutically acceptable excipients include binders (e.g., pregelatinized maize starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose), fillers (e.g., lactose, microcrystalline cellulose, or calcium phosphate), lubricants (e.g., magnesium stearate, talc, or silica), disintegrants (e.g., potato starch or sodium starch glycolate), or wetting agents (e.g., sodium lauryl sulfate). Tablets can be coated by methods well known in the art. For tablets, capsules, caplets, pellets, or granules for oral administration, a pH-sensitive enteric coating designed to control the release of the active ingredient may be used. Oral dosage forms also include modified-release formulations, such as immediate-release and sustained-release formulations. Examples of modified-release formulations include, for example, sustained-release (SR), extended-release (ER, XR, or XL), sustained or extended-release, controlled-release (CR), or continuous-release (CR or Contin, continuous-release) formulations used in the form of, for example, coated tablets, osmotic delivery devices, coated capsules, microencapsulated microspheres, agglomerated particles such as, for example, molecular sieve-type particles, or fine hollow permeable fiber bundles or chopped hollow permeable fibers agglomerated or held in a fibrous packet.Sustained-release compositions can be formulated, for example, in liposomes, or active compounds are protected by differentially degradable coatings, such as microencapsulation, multiple coatings, etc. Liposomal delivery systems include, for example, small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from various phospholipids, such as cholesterol, stearylamine, or phosphatidylcholine. For oral administration in capsule form, useful carriers or diluents include lactose and dried cornstarch.

[0139] Liquid preparations for oral administration can take the form of, for example, solutions, syrups, or suspensions, or can be conveniently provided as a dry product for constitution with water or other suitable vehicle before use. When aqueous suspensions and / or emulsions are administered orally, the compounds of the present application are suitably suspended or dissolved in an oily phase combined with an emulsifying and / or suspending agent. If desired, certain sweetening and / or flavoring and / or coloring agents may be added. Such liquid preparations for oral administration can be prepared by conventional means using pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats), emulsifying agents (e.g., lecithin or acacia), non-aqueous vehicles (e.g., almond oil, oily esters, or ethyl alcohol), and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid). Useful diluents include lactose and high molecular weight polyethylene glycols.

[0140] The compounds of the present application can also be freeze-dried and the resulting lyophilisates used, for example, in the preparation of injectable products.

[0141] The compounds of the present application can also be administered parenterally. Solutions of the compounds of the present application can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof, with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. Those skilled in the art will understand how to prepare appropriate formulations. For parenteral administration, sterile solutions of the compounds of the present application are usually prepared, and the pH of the solution is appropriately adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. For ocular administration, ointments or droppable liquids can be delivered by ocular delivery systems known in the art, such as applicators or eyedroppers. Such compositions can include mucus mimetics such as hyaluronic acid, chondroitin sulfate, hydroxypropylmethylcellulose, or polyvinyl alcohol, preservatives such as sorbic acid, EDTA, or benzyl chromium chloride, and conventional amounts of diluents or carriers. For pulmonary administration, a diluent or carrier is selected to be appropriate to allow the formation of an aerosol.

[0142] The compounds of the present application can be formulated for parenteral administration by injection, including using conventional catheterization or infusion. Injectable preparations can be provided in unit dosage form, for example, in ampoules or multi-dose containers, with added preservatives. The compositions can take the form of sterile suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. In all cases, the form must be sterile and fluid to the extent that easy syringability exists. Alternatively, the compounds of the present application are suitably in sterile powder form for reconstitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.

[0143] Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels and powders.

[0144] For intranasal administration or administration by inhalation, the compounds of the present application are conveniently delivered in the form of a solution, dry powder formulation, or suspension from a pump spray container that the patient presses or pumps, or as an aerosol spray formulation from a pressurized container or nebulizer. Aerosol formulations typically comprise a solution or fine suspension of the active agent in a physiologically acceptable aqueous or non-aqueous solvent and are usually provided in single or multi-dose form in a sterile, sealed container which may take the form of a cartridge or refill for use with a spray device.

[0145] Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein the active ingredient is formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerin. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0146] In some embodiments, the compounds of the present application can also be coupled to soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds of the present application can be coupled to biodegradable polymer classes useful for achieving controlled drug release, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, polyepsilon caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels.

[0147] In some embodiments, one or more compounds of the present application can also be coupled to a suitable antibody as a targetable drug carrier. For example, the compounds of the present application can be coupled to a suitable antibody to form an antibody-drug conjugate (ADC), for example, as described in Polakis, P., Pharmacol. Revs., 2016, 68, 3-19. Thus, the present application includes ADCs comprising one or more compounds of the present application. Antibodies generally can include any polypeptide comprising a framework region derived from an immunoglobulin or a fragment thereof that specifically binds to and recognizes an antigen, such as a carbohydrate, polynucleotide, lipid, or polypeptide. In some embodiments, the antibody specifically binds to a cancer antigen, for example, known cancer antigens CD22, CD33, CD30, HER2, mesothelin, Melan-A, CD19, CD20, CD79b, Trop2, HER3, MAGE, or MART-1. In some embodiments, the antibody specifically binds to an immune cell surface receptor, e.g., CD80, CD86, GMCSF-R, DC-SIGN, or CD36. In some embodiments, the antibody specifically binds to an aberrantly expressed protein characteristic of cancer. In some embodiments, the compounds of the present application can be coupled to an appropriate antibody via a linker to form an ADC. The linker may be or include a cleavable group that allows the compounds of the present application to be cleaved from the remainder of the complex in vivo by the biological environment. In some embodiments, the linker is cleaved within or near the desired target site of action. In some embodiments, the linker is non-cleavable.

[0148] In some embodiments, the compounds of the present application can be coupled to suitable viral, non-viral, or other vectors. Viral vectors can include retroviruses, lentiviruses, adenoviruses, herpes viruses, poxviruses, alphaviruses, vaccinia viruses, and / or adeno-associated viruses. Non-viral vectors can include nanoparticles, cationic lipids, cationic polymers, metal nanoparticles, nanorods, liposomes, micelles, microbubbles, cell-penetrating peptides, and / or lipospheres. Nanoparticles can include silica, lipids, carbohydrates, and / or other pharmaceutically acceptable polymers. While the compounds of the present application, including their pharmaceutically acceptable salts and solvates, are suitably used per se, they are generally administered in the form of a pharmaceutical composition in which one or more compounds of the present application (active ingredients) are associated with a pharmaceutically acceptable carrier. Depending on the mode of administration, the pharmaceutical composition comprises from about 0.05% to about 99% by weight, or from about 0.10% to about 70% by weight of the active ingredient (one or more compounds of the present application), and from about 1% to about 99.95% by weight, or from about 30% to about 99.90% by weight of a pharmaceutically acceptable carrier, all weight percentages being based on the total composition.

[0149] The compounds of the present application can be used alone or in combination with other known agents useful for treating diseases, disorders, or conditions mediated by or treatable by STING protein activation. When used in combination with other agents useful for treating diseases, disorders, or conditions mediated by or treatable by STING protein activation, in one embodiment, the compounds of the present application are administered simultaneously with those agents. As used herein, "co-administration" of two substances to a subject means providing each of the two substances so that they are both biologically active in an individual at the same time. The exact details of administration depend on the pharmacokinetics of the two substances in the presence of each other and can include administering the two substances within a few hours of each other, or even administering one substance within 24 hours of the other, if the pharmacokinetics are appropriate. Designing appropriate administration regimens is routine for those skilled in the art. In certain embodiments, the two substances are administered substantially simultaneously, i.e., within minutes of each other, or in a single composition containing both substances. In further embodiments of the present application, the drug combination is administered to a subject in a non-simultaneous manner. In one embodiment, a compound of the present application is administered with another therapeutic agent, either simultaneously or sequentially in separate unit dosage forms, or together in a single unit dosage form. Thus, the present application provides a single unit dosage form comprising one or more compounds of the present application (e.g., a compound of Formula I), an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0150] The dosage of the compounds of the present application can vary depending on many factors, including the pharmacodynamic properties of the compound, the mode of administration, the recipient's age, health, and weight, the nature and severity of symptoms, the frequency of treatment, and the type of concomitant treatment, if any, as well as the clearance rate of the compound in the treated subject. Those skilled in the art can determine appropriate dosages based on the above factors. The compounds of the present application can be initially administered at an appropriate dosage, which can be adjusted as necessary depending on the clinical response. The dosage is generally selected to maintain serum levels of the compounds of the present application between about 0.01 μg / cc and about 1000 μg / cc, or between about 0.1 μg / cc and about 100 μg / cc. Typically, the oral dosage of one or more compounds of the present application ranges from about 1 mg per day to about 1000 mg per day for adults, suitably between about 1 mg per day and about 500 mg per day, and more suitably between about 1 mg per day and about 200 mg per day. For parenteral administration, a typical dose is about 0.001 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For oral administration, a typical dose is about 0.001 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For administration in suppository form, a typical dose is about 0.1 mg / kg to about 10 mg / kg or about 0.1 mg / kg to about 1 mg / kg.

[0151] In one embodiment of the present application, the composition is formulated for oral administration and the compound is suitably in the form of tablets containing 0.25, 0.5, 0.75, 1.0, 5.0, 10.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 75.0, 80.0, 90.0, 100.0, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of active ingredient per tablet. The compounds of the present application can be administered in a once-daily, once-weekly, or once-monthly dose, or the total daily dose can be divided into two, three, or four doses per day.

[0152] For clarity, in the above, the term "a compound" also includes embodiments where one or more compounds are referred to.

[0153] Methods and Uses of the Disclosed Compounds The compounds of the present application have been shown to be capable of activating STING activity, such as STING protein activity.

[0154] Thus, the present application includes a method for activating STING in a cell, either in a biological sample or in a patient, comprising administering to the cell an effective amount of one or more compounds of the present application. The present application also includes the use of one or more compounds of the present application for activating STING in a cell, as well as the use of one or more compounds of the present application for preparing a medicament for activating STING in a cell. The present application further includes one or more compounds of the present application for use in activating STING in a cell.

[0155] Because the compounds of the present application have been shown to be capable of activating STING protein activity, the compounds of the present application are useful for treating diseases, disorders, or conditions by activating STING. Thus, the compounds of the present application are useful as pharmaceuticals. Thus, the present application includes the compounds of the present application for use as pharmaceuticals.

[0156] The present application also includes a method for treating a disease, disorder, or condition caused by activation of STING, comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of the present application.

[0157] The present application also includes the use of one or more compounds of the present application for treating a disease, disorder, or condition caused by activation of STING, as well as the use of one or more compounds of the present application for preparing a medicament for treating a disease, disorder, or condition caused by activation of STING. The present application further includes one or more compounds of the present application for use in treating a disease, disorder, or condition caused by activation of STING.

[0158] In one embodiment, the disease, disorder, or condition is a neoplastic disorder. Accordingly, the present application also includes a method of treating a neoplastic disorder, comprising administering a therapeutically effective amount of one or more compounds of the present application to a subject in need thereof. The present application also includes the use of one or more compounds of the present application for treating a neoplastic disorder, as well as the use of one or more compounds of the present application for preparing a medicament for treating a neoplastic disorder. The application further includes one or more compounds of the present application for use in treating a neoplastic disorder. In one embodiment, the treatment is an amount effective to ameliorate at least one symptom of a neoplastic disorder, e.g., inter alia, reduce cell proliferation or reduce tumor burden, in a subject in need of such treatment.

[0159] The compounds of the present application have been demonstrated to be effective against a panel of human tumor cell lines. Accordingly, in another embodiment of the present application, the disease, disorder, or condition treated by STING activation is cancer. Accordingly, the present application also includes a method of treating cancer, comprising administering a therapeutically effective amount of one or more compounds of the present application to a subject in need thereof. The present application also includes the use of one or more compounds of the present application for treating cancer, as well as the use of one or more compounds of the present application for preparing a medicament for treating cancer. The present application further includes one or more compounds of the present application for use in treating cancer. In one embodiment, the compound is administered to prevent cancer in a subject, such as a mammal, having a predisposition to cancer.

[0160] In one embodiment, the cancer is selected from cancer of the skin, blood, prostate, colorectal, pancreas, kidney, ovary, breast, e.g., mammary gland, liver, tongue, and lung. In another embodiment, the cancer is selected from leukemia, lymphoma, non-Hodgkin's lymphoma, and multiple myeloma. In a further embodiment of the application, the cancer is selected from leukemia, melanoma, lung cancer, colon cancer, brain cancer, ovarian cancer, breast cancer, prostate cancer, and kidney cancer.

[0161] In one embodiment, the disease, disorder, or condition treated by activation of STING is a disease, disorder, or condition associated with unregulated and / or abnormal cellular activity that is directly or indirectly affected by activation of STING. In another embodiment, the unregulated and / or abnormal cellular activity that is directly or indirectly affected by activation of STING is proliferative activity in a cell. Accordingly, the present application also includes a method for inhibiting proliferative activity in a cell, comprising administering to the cell an effective amount of one or more compounds of the present application. The present application also includes the use of one or more compounds of the present application for inhibiting proliferative activity in a cell, as well as the use of one or more compounds of the present application for preparing a medicament for inhibiting proliferative activity in a cell. The present application further includes one or more compounds of the present application for use in inhibiting proliferative activity in a cell.

[0162] The present application also includes a method for inhibiting unregulated and / or abnormal cellular activity that is directly or indirectly affected by a STING protein in a cell, either in a biological sample or in a subject, comprising administering to the cell an effective amount of one or more compounds of the present application. The present application also includes the use of one or more compounds of the present application for inhibiting unregulated and / or abnormal cellular activity that is directly or indirectly affected by a STING protein in a cell, as well as the use of one or more compounds of the present application for preparing a medicament for inhibiting unregulated and / or abnormal cellular activity that is directly or indirectly affected by a STING protein in a cell. The present application further includes one or more compounds of the present application for use in inhibiting unregulated and / or abnormal cellular activity that is directly or indirectly affected by a STING protein in a cell.

[0163] Accordingly, the present application also includes a method for treating a disease, disorder, or condition treatable by activation of STING, comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of the present application in combination with another known agent useful for treating the disease, disorder, or condition treatable by activation of STING. The present application also includes the use of one or more compounds of the present application in combination with another known agent useful for treating the disease, disorder, or condition treatable by activation of STING to treat the disease, disorder, or condition treatable by activation of STING, as well as the use of one or more compounds of the present application in combination with another known agent useful for treating the disease, disorder, or condition treatable by activation of STING to prepare a medicament for treating the disease, disorder, or condition treatable by activation of STING. The present application further includes one or more compounds of the present application in combination with another known agent useful for treating the disease, disorder, or condition treatable by activation of STING for use in treating the disease, disorder, or condition treatable by activation of STING. In one embodiment, the disease, disorder, or condition treatable by activation of STING is cancer, such as acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), melanoma, prostate cancer, endometrial cancer, schwannoma, mantle cell lymphoma, rhabdomyosarcoma, glioma, glioblastoma, B-ALL, T-ALL, lung cancer, gastric cancer, pancreatic cancer, and breast cancer.

[0164] In a further embodiment, the disease, disorder, or condition treatable by activation of STING is cancer, and one or more compounds of the present application are administered in combination with one or more additional cancer treatments, hi another embodiment, the additional cancer treatment is selected from radiation therapy, chemotherapy, targeted therapy, such as antibody therapy and small molecule therapy, such as tyrosine kinase inhibitors, immunotherapy, hormonal therapy, and anti-angiogenic therapy.

[0165] In some embodiments, the subject is a mammal, hi some embodiments, the subject is a human.

[0166] general synthesis The compounds disclosed herein can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, examples of which are described in the Examples below. [Example]

[0167] As shown in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures: While the general methods illustrate the synthesis of specific compounds of the present disclosure, it is understood that the following general methods, and other methods known to those of skill in the art, are applicable to all compounds, as well as each subclass and species of these compounds, as described herein.

[0168] Materials and Methods (Examples 1-19 and 29)

[0169] UPLC-MS Apparatus and Method

[0170] Equipment: Waters H-Class UPLC equipped with QSM, sample organizer, column heater, PDa UV detector and Qda mass spectrometer.

[0171] Column: Waters BEH C 18 Column, 100 x 2.1 mm, 1.7 μm, 130 Å pore size

[0172] Lcms_long method: Column temperature 40°C. UV absorption wavelength: 214 nm. MS range: 200-1250 Da. Mobile phase A: 0.1% TFA in water. Mobile phase B: 0.085% TFA in acetonitrile. Flow rate: 0.5 mL / min. Gradient: [Table 2]

[0173] Column: Waters BEH C 18Column, 50 x 2.1 mm, 1.7 μm, 130 Å pore size

[0174] Lcms_short method: Column temperature 40°C. UV absorption wavelength: 214 nm. MS range: 200-1250 Da. Mobile phase A: 0.1% TFA in water. Mobile phase B: 0.085% TFA in acetonitrile. Flow rate: 0.5 mL / min. Gradient: [Table 3]

[0175] Column: ACQUITY UPLC Protein BEH C4 Column, 300Å, 1.7μm, 2.1mm x 100mm

[0176] Lcms_long_C4 Method: Column temperature 40°C. UV absorption wavelength: 214 nm. MS range: 200-1250 Da. Mobile phase A: 0.1% TFA in water. Mobile phase B: 0.085% TFA in acetonitrile. Flow rate: 0.5 mL / min. Gradient: [Table 4]

[0177] HPLC Apparatus and Method

[0178] Equipment: Waters 2767 Autopure equipped with mass spectrometer. Mass spectrometer range: 200-3000 Da. UV detector wavelength: 214 nm.

[0179] Column: Phenomenex Luna C5 column, 250 x 21.2 mm, 10 μm particle size, 100 Å pore size.

[0180] Method: Flow rate of 20 mL / min. Mobile phase A: 0.05% TFA in water. Mobile phase B: 0.05% TFA in acetonitrile. Gradient: [Table 5]

[0181] Column: Phenomenex Luna 18 column, 250 x 30 mm, 10 μm particle size, 100 Å pore size.

[0182] Method: Flow rate of 30 mL / min. Mobile phase A: 0.05% TFA in water. Mobile phase B: 0.05% TFA in acetonitrile. Gradient: [Table 6] [Example]

[0183] Synthetic Scheme I [ka]

[0184] Preparation of 4-chloro-3-methoxy-5-nitrobenzamide (1.2) A suspension of methyl 4-chloro-3-methoxy-5-nitrobenzoate (1.1) (50.0 g, 204 mmol, 1.0 equiv.) in 30% aqueous ammonium hydroxide (733 mL) was stirred at 50° C. for 1.5 h (under pressure) and then at room temperature for 16 h. The mixture was then stirred at 50° C. for 24 h (open system). The reaction mixture was cooled to room temperature and filtered. The solid was washed with water (500 mL), diethyl ether (400 mL), and dried (50° C., 61 h) to give pure compound 1.2 (40.55 g, 176 mmol, 86%) as a yellow solid. 1 H NMR (DMSO-d6): δ4.02(s,3H), 7.78(brs,1H), 7.88(d,1H,J=1.7Hz), 8.05(d,1H,J=1.7Hz), 8.29(brs,1H). LCMS (2~100 ACN / H2O+0.1%FA): Tr=2.66 min; Purity=100%; [M+H + ] + =231.2.

[0185] Preparation of tert-butyl N-[(2E)-4-[(4-carbamoyl-2-methoxy-6-nitrophenyl)amino]but-2-en-1-yl]carbamate (1.3) To a suspension of tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (10.9 g, 58.5 mmol, 1.5 equiv.) and compound 1.2 (9.0 g, 39.0 mmol, 1.0 equiv.) in n-butanol (105 mL), DIPEA (21.5 mL, 123 mmol, 3.2 equiv.) was added. The reaction mixture was stirred at 120 °C for 16 h. The mixture was cooled to room temperature, then to 0 °C, and filtered. The solid was washed with cold ethanol (3 × 80 mL) and dried under vacuum to give pure compound 1.3 (13.6 g, 35.9 mmol, 92%) as a brick-red crystalline solid. 1 H NMR (DMSO-d6): δ1.35(s,9H), 3.40~3.52(m,2H), 3.87(s,3H), 4.05~4.09(m,2H), 5.49~5.58(m,2H), 6.56(br s,0.2H), 6.93(t,0.8H,J=5.3Hz), 7.32(br s,1H), 7.55(d,1H,J=1.6Hz), 7.74(d,1H,J=6.0Hz), 8.01(br s,1H), 8.18(d,1H,J=1.9Hz). LCMS (2~100 ACN+0.1%FA / H2O+0.1%FA): Tr=2.85 min; Purity=100%; [M+Na + ] + =403.4.

[0186] Preparation of 4-{[(2E)-4-aminobut-2-en-1-yl]amino}-3-methoxy-5-nitrobenzamide hydrochloride (1.4) To a suspension of compound 1.3 (12.4 g, 32.5 mmol, 1.0 equiv.) in methanol (30.9 mL) was added dropwise 4 M HCl in dioxane (61.8 mL, 247 mmol, 7.6 equiv.). The resulting solution was stirred at room temperature for 1 h. The formed solid was filtered, washed with diethyl ether (3 × 100 mL), and dried under reduced pressure (40 °C, 16 h) to give crude compound 1.4 (10.3 g, 32.5 mmol, quantitative) as an orange solid. 1H NMR (DMSO-d6): δ3.36~3.42(m,2H), 3.89(s,3H), 4.16~4.18(m,2H), 4.71(br s,1H), 5.59~5.66(m,1H), 5.84~5.90(m,1H), 7.37(br s,1H), 7.59(d,1H,J=1.9Hz), 8.00~8.07(m,4H), 8.21(d,1H,J=1.9Hz). LCMS (2~100 ACN / H2O+0.1%FA): Tr=2.12 min; Purity=10%; [M-HCl+H + ] + =281.3.

[0187] Preparation of 4-chloro-3-hydroxy-5-nitrobenzamide (1.5) To a solution of compound 1.2 (31.5 g, 136 mmol, 1.0 equiv.) in dichloromethane (400 mL) cooled to 0° C., boron tribromide (1 M in DCM, 545 mL, 545 mmol, 4.0 equiv.) was added over 20 min, and the mixture was stirred at 40° C. for 42 h. The reaction mixture was allowed to reach room temperature, poured into 500 mL of ice / water, and stirred for 10 min (until the ice melted). The precipitate was filtered, washed twice with water and twice with n-pentane, and dried under reduced pressure (50° C., 16 h) to give a white solid (41.4 g). The residue was triturated in water (400 mL) for 1 h and filtered. The filter cake was washed with water (500 mL) and n-pentane (2×500 mL) and dried under vacuum to give a white solid (25.3 g). The residue was triturated and triturated in water (400 mL) for 2 hours, then filtered. The filter cake was washed with water (500 mL) and n-pentane (2×500 mL) and dried under vacuum to give crude compound 1.5 (23.5 g, 108.5 mmol, 80%) as a white solid. 1 H NMR (DMSO-d6): δ7.67(br s,1H), 7.72(d,1H,J=1.8Hz), 7.93(d,1H,J=1.9Hz), 8.18(br s,1H), 11.53(br s,1H). LCMS(2~100 ACN+0.1%AF / H2O+0.1%AF):Tr=2.49min;Purity=90.58%;[M+H + ] + =217.2.

[0188] Preparation of tert-butyl 4-(5-carbamoyl-2-chloro-3-nitrophenoxy)butanoate (1.6) To a solution of compound 1.5 (23.5 g, 108.5 mmol, 1.0 equiv.) in acetonitrile (285 mL) was added tert-butyl 4-bromobutanoate (48.4 g, 38.5 mL, 217 mmol, 2.0 equiv.), followed by potassium carbonate (30.0 g, 217 mmol, 2.0 equiv.). The resulting solution was stirred under reflux. After 30 minutes, a solid appeared (no further stirring). The solid was crushed, acetonitrile (200 mL) was added, and the reaction mixture was refluxed for 18 hours. The reaction mixture was concentrated under reduced pressure, diluted with water (700 mL) and ethyl acetate (500 mL), stirred for 10 minutes to dissolve the solid, and extracted with ethyl acetate (2 x 500 mL). The organic phase was washed with water (300 mL), brine (300 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a light brownish solid (60.2 g). The residue was suspended in n-pentane (30 mL) and sonicated for 10 min (large chunks of solid were first broken up with a spatula to give a homogeneous powder). The suspension was filtered through a sintered glass funnel and washed with n-pentane (4 × 20 mL). The resulting solid was dried under reduced pressure to give pure compound 1.6 (36 g, 100 mmol, 92%) as a white solid. 1 H NMR (DMSO-d6): δ1.10(s,9H), 1.98~2.05(m,2H), 2.43(t,2H,J=7.4Hz), 4.25(t,2H,J=6.2Hz), 7.78(br s,1H), 7.87(d,1H,J=1.8Hz), 8.05(d,1H,J=1.8Hz), 8.29(br s,1H). LCMS (2~100 ACN / H2O+0.1%AF): Tr=3.11 min; Purity=96.02%; [M-tBu+H + ] + =303.2.

[0189] Preparation of tert-butyl 4-(5-carbamoyl-2-{[(2E)-4-[(4-carbamoyl-2-methoxy-6-nitrophenyl)amino]but-2-en-1-yl]amino}-3-nitrophenoxy)butanoate (1.7) To a stirred solution of compound 1.4 (1.96 g, 5.57 mmol, 1.0 equiv.) in n-butanol (37.8 mL), DIPEA (4.61 mL, 27.9 mmol, 5.0 equiv.) and sodium bicarbonate (937 mg, 11.1 mmol, 2.0 equiv.) were added, and the mixture was stirred at room temperature for 10 minutes. Compound 1.6 (2.00 g, 5.57 mmol, 1.0 equiv.) was then added, and the reaction mixture was stirred at 120 °C for 22 hours. The reaction mixture was quenched with water (150 mL) at room temperature and extracted with a mixture of dichloromethane / methanol (9 / 1, 3 × 150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to give crude compound 1.7 (3.36 g, 5.58 mmol, assumed quantitative) as an orange oil. Crude compound 1.7 was used directly in the next step. LCMS (2~100 ACN / H2O+0.1%AF): Tr=3.01 min, purity=61.46%, [M+H + ] + =603.5.

[0190] Preparation of tert-butyl 4-(3-amino-2-{[(2E)-4-[(2-amino-4-carbamoyl-6-methoxyphenyl)amino]but-2-en-1-yl]amino}-5-carbamoylphenoxy)butanoate (1.8) To a solution of crude impure compound 1.7 (3.34 g, 5.54 mmol, 1.0 equiv.) in ethanol (45 mL) was added dichlorostannane dihydrate (10.0 g, 44.3 mmol, 8.0 equiv.). The mixture was then heated to 60° C. and stirred for 3 h. The reaction was cooled, diluted with saturated aqueous potassium carbonate (100 mL) and water (100 mL), and extracted with a mixture of dichloromethane / methanol (85:15, 3×250 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a white solid (2.67 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 80 g) using dichloromethane / methanol (98:2 to 80:20 in 50 min) to give pure compound 1.8 (1.28 g, 2.36 mmol, 43% over two steps) as a yellow solid. 1 H NMR (DMSO-d6): δ1.38(s,9H), 1.89~1.93(m,2H), 2.39(t,2H,J=7.4Hz), 3.48~3.52(m,4H), 3.73(s,3H), 3.79(dt,2H,J=26.6,6.8 Hz), 3.93(t,2H,J=6.2Hz), 4.64(d,4H,J=9.7Hz), 5.62~5.71(m,2H), 6.76(dd,2H,J=8.4,1.8Hz), 6.85(t,2H,J=1.7Hz), 6.96(br s,2H), 7.60(br s,2H). LCMS (2~100 ACN / H2O+0.1%AF):Tr=2.45min;purity=99.23%;[M+H + ] + =543.6.

[0191] Preparation of tert-butyl 4-({5-carbamoyl-1-[(2E)-4-[5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-amido)-7-methoxy-1H-1,3-benzodiazol-1-yl]but-2-en-1-yl]-2-(1-ethyl-3-methyl-1H-pyrazol-5-amido)-1H-1,3-benzodiazol-7-yl}oxy)butanoate (1.9) To a solution of compound 1.8 (5.60 g, 10.3 mmol, 1.0 equiv) in N,N-dimethylformamide (110 mL) at 0 °C, compound 1.11 (0.2 M in dioxane, 20.6 mL, 10.3 mmol, 1.0 equiv) was added and the reaction mixture was stirred for 15 minutes. Compound 1.11 (0.2 M in dioxane, 8.26 mL, 4.13 mmol, 0.4 equiv) was added and the reaction mixture was stirred for 15 minutes. Compound 1.11 (0.2 M in dioxane, 4.13 mL, 2.06 mmol, 0.2 equiv) was added and the reaction mixture was stirred for 15 minutes. Compound 1.11 (0.2 M in dioxane, 8.26 mL, 4.13 mmol, 0.4 equiv) was added and the reaction mixture was stirred for 15 minutes. EDC·HCl (4.95 g, 25.8 mmol, 2.5 equiv.) followed by triethylamine (7.17 mL, 51.6 mmol, 5.0 equiv.) was added to the reaction at 0 °C, warmed, and stirred at room temperature for 18 h. Triethylamine (7.17 mL, 51.6 mmol, 5.0 equiv.) and EDC·HCl (4.95 g, 25.8 mmol, 2.5 equiv.) were added, and the reaction mixture was stirred at room temperature for 24 h. The reaction was quenched with a mixture of water / saturated aqueous ammonium chloride (3:1, 400 mL) and extracted with a mixture of dichloromethane / methanol (3:1, 3 × 400 mL). The combined organic layers were washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give an orange oil (32 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 330 g) using dichloromethane / methanol (90 / 10 for 15 min, then 80 / 20 for 25 min) to give impure compound 1.9 (8.03 g) as a yellow solid. The residue was triturated in acetonitrile (2 × 125 mL) at 45 °C for 2 h and then filtered. The solid was dissolved in a mixture of dichloromethane / methanol (3:1, 700 mL) and washed with water (2 × 400 mL). The aqueous layer was extracted with a mixture of dichloromethane / methanol (3:1, 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated, and dried under vacuum (40 °C, 16 h) to give pure compound 1.9 (6.00 g, 6.94 mmol, 67%) as a beige solid. 1H NMR (DMSO-d6): δ1.26(t,6H,J=7.0Hz), 1.39(s,9H), 1.76~1.83(m,2H), 2.09(s,3H), 2.10(s,3H), 2.25(t,2H,J=7.2Hz), 3.72(s,3H), 3 .98(t,2H,J=6.1Hz), 4.52(q,4H,J=7.0Hz), 4.90~4.93(m,4H), 5.76~5.89(m,2H), 6.50(d,2H,J=5.3Hz), 7.30(d,2H,J=5.3Hz), 7.34(br s,2H), 7.64(s,2H), 7.95(br s,2H), 12.81(br s,2H). LCMS (2~100 ACN / H2O+0.1%AF):Tr=9.55min;purity=94.58%;[M+H + ] + =865.46.

[0192] Preparation of Compound 1.12 To a suspension of compound 1.9 (400 mg, 0.46 mmol) in THF (4 mL) at room temperature, HCl (4 M in dioxane, 3.0 mL, 12.0 mmol) was added, and the resulting mixture was stirred at room temperature for 24 hours. Complete conversion to compound 1.12 was confirmed by monitoring with UPLC-MS. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the resulting precipitate was isolated by centrifugation. The supernatant was then removed, and the resulting pellet was resuspended in cold diethyl ether (100 mL) and centrifuged. This step was repeated twice. Finally, the pellet was dried in vacuo to give compound 1.12 as the HCl salt, a beige solid (386.5 mg; 0.44 mmol, 95%). The product was used in the next step without further purification. UPLC-MS retention time: 3.26 min (lcms_long), [M+H + ] + =809.51.

[0193] Preparation of Compound 1.13 To a suspension of compound 1.12 (250.1 mg, 0.28 mmol, 1.0 equiv., HCl salt) in DMF (1 mL) was added DIEA (148.7 μL, 0.85 mmol, 3.0 equiv.) at 40 °C, followed by the addition of N-hydroxysuccinimide (161 mg, 1.4 mmol, 5.0 equiv.) and N,N'-diisopropylcarbodiimide (212.0 mg, 1.68 mmol, 6.0 equiv.). The resulting mixture was stirred for 2 h. Complete conversion to compound 1.13 was confirmed by UPLC-MS monitoring. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the formed precipitate was isolated by centrifugation. The supernatant was removed, and the formed pellet was resuspended in cold diethyl ether (100 mL) and centrifuged. This step was repeated twice. Finally, the pellet was dried in vacuo to give compound 1.13 as a beige solid (220.7 mg, 0.24 mmol, 87%). The product was used in the next step without further purification. UPLC-MS retention time: 4.40 min (lcms_long), [M+H + ] + 906.54.

[0194] Preparation of Compound IA [ka]

[0195] General Method A: To a solution of compound 1.13 (45.0 mg, 0.05 mmol, 1.0 equiv.) in DMF (3 mL) at room temperature, hydrazine hydrate (20 μL, 0.49 mmol, 10.0 equiv.) was added. The resulting mixture was stirred for 1 h. Complete consumption of the starting material and formation of compound IA were confirmed by UPLC-MS monitoring. The reaction mixture was diluted with water, filtered through a 0.22 μm syringe filter, and directly injected into RP-HPLC for purification (Phenomenex Luna C18 250 × 30 mm column, 30 mL / min flow rate, 0.05% TFA, 30–45% MeCN / HO gradient over 35 min, 50 min total run time). Fractions containing pure product were pooled, frozen, and lyophilized to afford compound IA as a white solid (25 mg, 0.03 mmol, 60%). UPLC-MS retention time: 3.73 min broad peak (lcms_long), [M+H + ] + =823.52.

[0196] Preparation of compound IB [ka]

[0197] Compound IB was prepared according to modified general method A using monomethylhydrazine, with MeCN as the reaction solvent and the reaction carried out at 4 °C, to give compound IB as a beige solid. UPLC-MS retention time: 3.90 min (1cms_long), [M+H + ] + =837.59.

[0198] Preparation of compound IC [ka]

[0199] General Method B: To a solution of compound 1.13 (30 mg, 0.033 mmol, 1.0 equiv.) in MeCN (1 mL) at room temperature, 1-Boc-1-methylhydrazine (48.4 mg, 0.33 mmol, 10.0 equiv.) was added. The resulting mixture was stirred at 40° C. for 16 hours. Complete consumption of the starting material and the formation of the intermediate compound IC-Boc were confirmed by monitoring with UPLC-MS. The reaction mixture was cooled to room temperature, and then trifluoroacetic acid (400 μL) was added, followed by HCl (4 M in dioxane, 200 μL). The resulting mixture was stirred at room temperature for 1 hour, during which time the mixture became cloudy. Complete Boc removal and formation of the desired compound IC were confirmed by monitoring with UPLC-MS. The reaction mixture was then further diluted to 7 mL with water, filtered through a 0.22 μm syringe filter, and directly injected into RP-HPLC for purification (Phenomenex Luna C18 250 x 30 mm column, 30 mL / min flow rate, 0.05% TFA, 30-45% MeCN / HO gradient over 35 min, 50 min total run time). Fractions containing pure product were pooled, frozen, and lyophilized to afford compound IC as a white solid (3.5 mg, 0.0042 mmol, 12%). UPLC-MS retention time: 3.87 min (lcms_long), [M+H + ] + =837.59.

[0200] Preparation of compound 1-D [ka]

[0201] General Method C: To a solution of compound 1.13 (170 mg, 0.187 mmol, 1.0 equiv.) in DMF (3 mL) at room temperature, N,N'-dimethylhydrazine dihydrochloride (42.4 mg, 0.32 mmol, 1.7 equiv.) was added, followed by DIEA (130.4 μL, 0.75 mmol, 4.0 equiv.). The resulting mixture was stirred at room temperature for 5 min. Complete consumption of the starting material and formation of the desired compound ID were confirmed by UPLC-MS monitoring. The reaction mixture was further diluted to 6 mL with water, filtered through a 0.22 μm syringe filter, and directly injected onto RP-HPLC for purification (Phenomenex Luna C18 250 x 30 mm column, 30 mL / min flow rate, 0.05% TFA, 30–45% MeCN / HO gradient over 35 min, 50 min total run time). Fractions containing pure product were pooled, frozen, and lyophilized to give compound ID as a white solid (107.4 mg, 0.12 mmol, 67%). UPLC-MS retention time: 4.01 min (lcms_long), [M+H + ] + =851.56.

[0202] Preparation of compounds IE and IF [ka]

[0203] Compounds IE and IF were both isolated as beige solids from one reaction mixture following a modified general procedure B using tert-butyl (4-(hydrazinylmethyl)phenyl)carbamate and carrying out the reaction at room temperature.

[0204] Compound IE (main product): UPLC-MS retention time: 3.68 min (lcms_long), [M+Na + ] + =950.55.

[0205] Compound IF (trace product): UPLC-MS retention time: 3.78 min (lcms_long), [M+Na + ] + =950.55.

[0206] Preparation of compound IG [ka]

[0207] Compound IG was prepared according to general procedure C, using hydroxylamine hydrochloride (2.0 equiv.) and N-methylmorpholine (4.0 equiv.) instead of DIEA. UPLC-MS retention time: 3.66 min (lcms_long), [M+H + ] + =824.50 m / z.

[0208] Preparation of Compound IH [ka]

[0209] Compound IH was prepared according to general procedure C, using N-methylhydroxylamine hydrochloride (2.0 equiv.) and N-methylmorpholine (4.0 equiv.) instead of DIEA. UPLC-MS retention time: 4.02 min (1cms_long), [M+H + ] + =838.55 m / z.

[0210] Preparation of Compound II [ka]

[0211] Compound II was prepared according to general procedure C, using N,O-dimethylhydroxylamine hydrochloride (2.0 equiv.) and N-methylmorpholine (4.0 equiv.) instead of DIEA. UPLC-MS retention time: 4.36 min (1cms_long), [M+H + ] + =852.55.

[0212] Preparation of Compound IJ [ka]

[0213] Compound IJ was prepared as a white solid following a modified general procedure B using tert-butyl (2-hydrazinoethyl)carbamate, with the reaction carried out at room temperature. This compound was isolated as the only product of the reaction. UPLC-MS retention time: 3.71 min (1cms_long), [M+H + ] + =866.54.

[0214] Preparation of compound IK [ka]

[0215] Compound IK was prepared as a white solid according to a modified general procedure B using tert-butylpyrazin-1-ylcarbamate, with the reaction carried out at room temperature. UPLC-MS retention time: 3.74 min (1cms_long), [M+H + ] + =892.64. [Example]

[0216] Synthetic Scheme II [ka]

[0217] Preparation of methyl 4-{[(2E)-4-{[(tert-butoxy)carbonyl]amino}but-2-en-1-yl]amino}-3-methoxy-5-nitrobenzoate (2.1) To a solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate (1.1) (1.00 g, 4.07 mmol, 1.00 equiv.) and tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (834 mg, 4.48 mmol, 1.10 equiv.) in tetrahydrofuran (15 mL), potassium carbonate (1.13 g, 8.14 mmol, 2.00 equiv.) was added at room temperature. The reaction mixture was stirred at 60 °C for 64 h. The resulting mixture was concentrated in vacuo. The residue was redissolved in ethyl acetate (100 mL) and washed with water (2 × 50 mL) and saturated aqueous sodium chloride solution (100 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated in vacuo to give pure compound 2.1 (1.61 g, 4.07 mmol, quantitative) as an orange solid. 1 H NMR (DMSO-d6): 1.34(s,9H), 3.42~3.50(m,2H), 3.83(s,3H), 3.89(s,3H), 4.11~4.13(m,2H), 5.52~5.53(m,2H), 6.56(br s,0.1H), 6.92(t,0.9H,J=5.4Hz), 7.43(d,1H,J=1.8Hz), 8.01(t,1H,J=6.1Hz), 8.16(t,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%FA): Tr=3.23 min; Purity=98.1%; [M+Na + ]=418.4.

[0218] Preparation of methyl 4-{[(2E)-4-aminobut-2-en-1-yl]amino}-3-methoxy-5-nitrobenzoate hydrochloride (3.1) To a suspension of compound 2.1 (3.25 g, 7.23 mmol, 1.00 equiv.) in MeOH (7.15 mL) was added HCl (4 N in dioxane) (14.3 mL, 57.2 mmol, 7.90 equiv.) dropwise at room temperature. The reaction mixture was stirred at room temperature for 1.5 h. The precipitate was filtered and washed with diethyl ether (3 × 20 mL). The filtrate was filtered again and washed with diethyl ether (10 mL). The solids were combined and dried under reduced pressure to give pure crude compound 3.1 (2.38 g, 7.11 mmol, 98%) as an orange solid. 1H NMR(DMSO-d6):3.38~3.42(m,2H), 3.84(s,3H), 3.90(s,3H), 4.21(t,2H,J=5 .8Hz), 5.59~5.66(m,1H), 5.83~5.90(m,1H), 7.46(d,1H,J=1.8Hz), 7.99(br s,3H), 8.13(t,1H,J=6.4Hz), 8.19(d,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%FA): Tr=2.43 min; Purity=100%; [M-HCl+H + ]=296.4.

[0219] Preparation of 4-chloro-3-[3-(morpholin-4-yl)propoxy]-5-nitrobenzamide (3.2) A mixture of compound 1.5 (1.00 g, 4.62 mmol, 1.00 equiv.), 4-(3-chloropropyl)morpholine (907 mg, 5.54 mmol, 1.20 equiv.), and potassium carbonate (830 mg, 6.00 mmol, 1.30 equiv.) in N,N-dimethylformamide (6.00 mL) was stirred at 70 °C for 16 h. The solvent was removed in vacuo to give a yellow solid (2.71 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 80 g) using dichloromethane / methanol (100:0 to 85:15 over 30 min, then 85:15 for 10 min) to give pure compound 3.2 (1.17 g, 3.40 mmol, 74%) as a pale yellow solid. 1 H NMR (DMSO-d6): δ1.92~1.98(m,2H), 2.35~2.37(m,4H), 2.45(t,2H,J=7.1Hz), 3.56(t,4H,J=4.6Hz), 4.28(t,2H,J=6.3Hz), 7.77(br s,1H), 7.87(d,1H,J=1.7Hz), 8.04(d,1H,J=1.7Hz), 8.27(br s,1H). LCMS (2~100 ACN / H2O+0.1%AF): Tr=2.37min; Purity=100%; [M+H + ]=344.3.

[0220] Preparation of 4-{[(2E)-4-({4-carbamoyl-2-[3-(morpholin-4-yl)propoxy]-6-nitrophenyl}amino)but-2-en-1-yl]amino}-3-methoxy-5-nitrobenzamide (3.3) To a stirred solution of compound 3.1 (1.06 g, 3.20 mmol, 1.10 equiv) and compound 3.2 (1.00 g, 2.91 mmol, 1.00 equiv) in tetrahydrofuran (10.7 mL) was added potassium carbonate (804 mg, 5.82 mmol, 2.00 equiv) and DIPEA (1.68 mL, 10.2 mmol, 3.49 equiv). The reaction mixture was stirred at 100 °C for 3 days. The reaction mixture was cooled to room temperature, diluted with water (40 mL), and extracted with a mixture of dichloromethane / methanol (8 / 2, 3 × 40 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to give an orange solid (2.17 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 80 g) using dichloromethane / methanol (98:2 for 10 min, 92:8 for 20 min, then 92:8 for 10 min) to give pure compound 3.3 (1.17 g, 1.95 mmol, 67%) as an orange solid. 1 H NMR (DMSO-d6): δ1.85~1.91(m,2H), 2.32~2.34(m,4H), 2.38(t,2H,J=7.1Hz), 3.55(t,4H,J=4.5Hz ), 3.80(s,3H), 3.84(s,3H), 4.00(t,2H,J=6.3Hz), 4.07~4.11(m,4H), 5.52~5.62(m,2H), 7.30(br s,1H), 7.35(d,1H,J=1.8Hz), 7.47(d,1H,J=1.8Hz), 7.73(t,1H,J=6.2Hz), 7.95~7.98(m,2H), 8.10(d,1H,J=1.8Hz), 8.14(d,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%AF): Tr=2.59 min; Purity=100%; [M+H + ]=603.5.

[0221] Preparation of methyl 3-amino-4-{[(2E)-4-({2-amino-4-carbamoyl-6-[3-(morpholin-4-yl)propoxy]phenyl}amino)but-2-en-1-yl]amino}-5-methoxybenzoate (3.4) To a stirred solution of compound 3.3 (11.7 g, 19.5 mmol, 1.00 equiv) in MeOH (305 mL) was added sodium dithionite (33.9 g, 195 mmol, 10.0 equiv) dissolved in water (105 mL) at 0 °C. To this stirred mixture was added 30% aqueous ammonia (19.4 mL, 146 mmol, 7.50 equiv) at 0 °C. The mixture was warmed to room temperature and stirred for 6 h. The reaction mixture was quenched with water (500 mL). Dichloromethane was added to the mixture (700 mL). The layers were separated, and the aqueous layer was extracted with a mixture of dichloromethane / methanol (9:1, 2 × 300 mL). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give pure compound 3.4 (7.34 g, 13.5 mmol, 69%) as a brown foam, which was used crude in the next step. 1 H NMR (DMSO-d6): δ1.82~1.88(m,2H), 2.32~2.34(m,4H), 2.40(t,2H,J=7.0Hz), 3.51 ~3.56(m,6H), 3.60~3.62(m,2H), 3.73(s,3H), 3.76(s,3H), 3.80(t,1H,J=7.1Hz), 3 .95(t,2H,J=6.2Hz), 4.07(t,1H,J=7.0Hz), 4.65(s,2H), 4.79(s,2H), 5.59~5.70( m,2H), 6.76(d,1H,J=1.8Hz), 6.81(d,1H,J=1.8Hz), 6.84(d,1H,J=1.8Hz), 6.95(br s,1H), 7.01(d,1H,J=1.8Hz), 7.60(br s,1H). LCMS (2~100 ACN / H2O+0.1%AF): Tr=2.22 min; Purity=100%; [M+H + ]=543.5.

[0222] Preparation of methyl 1-[(2E)-4-[5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-amido)-7-[3-(morpholin-4-yl)propoxy]-1H-1,3-benzodiazol-1-yl]but-2-en-1-yl]-2-(1-ethyl-3-methyl-1H-pyrazol-5-amido)-7-methoxy-1H-1,3-benzodiazole-5-carboxylate (3.5) To a solution of compound 3.4 (2.00 g, 3.69 mmol, 1.00 equiv) in N,N-dimethylformamide (40 mL) was added compound 1.11 (0.2 M in dioxane, 18.4 mL, 3.69 mmol, 1.00 equiv) dropwise at 0 °C, and the reaction mixture was stirred for 30 minutes. Compound 1.11 (0.2 M in dioxane, 7.37 mL, 1.47 mmol, 0.40 equiv) was added dropwise at 0 °C, and the reaction mixture was stirred for 15 minutes. Compound 1.11 (0.2 M in dioxane, 3.69 mL, 0.74 mmol, 0.20 equiv) was added dropwise at 0 °C, and the reaction mixture was stirred for 15 minutes. Compound 1.11 (0.2 M in dioxane, 7.37 mL, 1.47 mmol, 0.40 equiv) was added dropwise at 0 °C, and the reaction mixture was stirred for 2 hours. EDC·HCl (1.77 g, 9.21 mmol, 2.50 equiv) followed by triethylamine (2.56 mL, 18.4 mmol, 5.00 equiv) were added to the reaction at 0 °C. The reaction mixture was stirred at room temperature for 17 h. EDC·HCl (0.18 g, 0.92 mmol, 0.25 equiv) and triethylamine (0.064 mL, 0.46 mmol, 0.13 equiv) were added at room temperature. The reaction mixture was stirred at room temperature for 6 h. The reaction was quenched with a mixture of water / saturated aqueous ammonium chloride (3:1, 80 mL) and extracted with a mixture of dichloromethane / methanol (3:1, 3 × 80 mL). The combined organic phases were washed with water (100 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give an orange oil. The residue was purified by flash chromatography on silica gel (50 μm amorphous, 220 g) using dichloromethane / methanol (96:4 for 5 min, 80:20 for 30 min, then 80:20 for 25 min) as the eluent to give impure compound 3.5 (1.83 g) as a beige solid. The residue was triturated in dichloromethane (40 mL) to give pure compound 3.5 (1.74 g, 2.01 mmol, 55%) as a white solid. 1H NMR (DMSO-d6): δ1.27~1.32(m,6H), 1.60~1.66(m,2H), 2.11(s,3H), 2.13(s,3H), 2.17~2.19(m,4H), 2.23(t,2H,J=7.2Hz), 3.45(t,4H,J=4. 5Hz), 3.65(s,3H), 3.67~3.90(m,5H), 4.51~4.58(m,4H), 4.90~4.92(m,4H), 5.75~5.87(m,2H), 6.54(m,2H), 7.23(d,2H,J=5.2Hz), 7.31(br s,1H), 7.63(s,1H), 7.77(d,1H,J=1.2Hz), 7.92(br s,1H), 12.82(br s,1H), 12.86(br s,1H). LCMS (2~100 ACN / H2O+0.1%AF): Tr=7.06 min; Purity=94.95%; [M+H + ]=865.75.

[0223] Preparation of Compound 3.6 To a solution of compound 3.5 (400 mg, 0.46 mmol, 1.00 equiv.) in a mixture of MeOH (3.2 mL), THF (6.5 mL), and water (1.6 mL) was added NaOH (5 M aqueous solution, 920 μg, 4.46 mmol, 10 equiv.). The resulting mixture was stirred at 40° C. for 1 h. Complete consumption of the starting material and formation of the desired compound 3.6 were confirmed by UPLC-MS monitoring. HCl (1 M aqueous HCl) was then added to the reaction mixture until pH 6 was reached. The cloudy mixture was diluted with water, frozen, and lyophilized. The resulting solid was triturated in 90% DCM, 9% MeOH, 1% DIEA. The solid was removed by filtration, and the effluent was concentrated in vacuo to give compound 3.6 as a beige solid (356 mg, 0.41, 91%). UPLC-MS retention time: 4.12 min (1cms_long), [M+H + ] + =851.56. UPLC-MS retention time: 4.12 min (lcms_long), [M+H + ] + =851.56.

[0224] Preparation of Compound 3.7 To a suspension of compound 3.6 (100 mg, 0.117 mmol, 1.0 equiv.) in DMF (1 mL) was added DIEA (61.5 μL, 0.35 mmol, 3.0 equiv.) at 40 °C, followed by the addition of N-hydroxysuccinimide (67 mg, 0.58 mmol, 5.0 equiv.) and N,N'-diisopropylcarbodiimide (88.3 mg, 0.7 mmol, 6.0 equiv.). The resulting mixture was stirred for 2 h. Complete conversion to compound 3.7 was confirmed by UPLC-MS monitoring. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the formed precipitate was isolated by centrifugation. The supernatant was removed, and the formed pellet was resuspended in cold diethyl ether (100 mL) and centrifuged. This step was repeated twice. Finally, the pellet was dried in vacuo to give compound 3.7 as a beige solid (90.0 mg, 0.094 mmol, 80%). The product was used in the next step without further purification. UPLC-MS retention time: 4.41 min (lcms_long), [M+H + ] + =948.62.

[0225] Preparation of Compound II-A [ka]

[0226] Compound II-A was prepared using anhydrous hydrazine according to general method A to give compound II-A as a beige solid. UPLC-MS retention time: 3.52 min broad (lcms_long), 3.41 min sharp (lcms_long_C4), [M+H + ] + =865.65.

[0227] Preparation of Compound II-B [ka]

[0228] Compound II-B was prepared using 1-Boc-1-methylhydrazine according to the modified general method B, with the addition of 4-dimethylaminopyridine (1 equivalent) as a catalyst and the reaction temperature increased to 60 °C, to give compound II-B as a beige solid. UPLC-MS retention time: 3.37 min (lcms_long_C4), [M+H + ] + =879.65.

[0229] Preparation of compound II-C [ka]

[0230] Compound II-C was prepared as a beige solid using monomethylhydrazine according to modified general procedure A, where the reaction solvent was MeCN and the reaction was carried out at 4 °C. UPLC-MS retention times: 3.73 min broad (lcms_long), 3.53 min (lcms_long_C4), [M+H + ] + =879.63.

[0231] Preparation of Compound II-D [ka]

[0232] Compound II-D was prepared according to general procedure C using N,N'-dimethylhydrazine dihydrochloride to give a beige solid. UPLC-MS retention time: 3.06 min broad (lcms_short); [M+H + ] + =893.64. [Example]

[0233] Synthetic Scheme III [ka]

[0234] Preparation of Compound 4.2 To a stirred solution of compound 1.3 (300 mg, 0.79 mmol) in 10 mL of MeOH in a 50 mL round-bottom flask was added NaSO (14 equiv., 1.926 g, 11.04 mmol) as a solution in 10 mL of water. The reaction mixture was stirred for 15 minutes, and then NaHCO (43 equiv., 2.85 g, 33.93 mmol) was added. The reaction was judged complete after 10 minutes by monitoring with UPLC-MS. The reaction mixture was diluted with water (100 mL), extracted twice with ethyl acetate (100 mL), washed with water (100 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give crude compound 4.2 (225.83 mg, 0.644 mmol, 82%). The product was used in the next step without further purification. UPLC-MS retention time: 2.41 min (lcms_short); [M+H + ] + =351.3.

[0235] Preparation of compound 4.3 To a stirred solution of compound 4.2 (180.6 mg, 0.5156 mmol) in 5 mL of DMF in a 20 mL glass scintillation vial, 0.058 mL of a 0.5 g / mL solution of compound 1.11 (28.8 mg, 0.148 mmol) in dioxane was added dropwise at 0 °C. After 15 min, a second equivalent of compound 1.11 was added, followed by a third equivalent 15 min later. Reaction monitoring by UPLC-MS indicated 87% conversion to the thiourea intermediate, so an additional 0.030 mL of compound 1.11 solution was added. After complete conversion to the thiourea intermediate, the reaction was warmed to room temperature and EDC·HCl (1.3 equiv., 128.5 mg, 0.67 mmol) was added as a solution in 3 mL of DMF, followed by DIEA (3 equiv., 0.27 mL, 1.55 mmol). The reaction was stirred overnight at room temperature. After 16 hours, monitoring by UPLC-MS indicated 88% conversion. An additional 30 mg of EDC·HCl (0.3 equiv., 0.156 mmol) was added. Five minutes after the second addition of EDC, the reaction was complete. The reaction mixture was diluted with water (100 mL), extracted twice with ethyl acetate (100 mL), then washed with water (100 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give crude compound 4.3. No yield was obtained in this step. UPLC-MS retention time: 3.58 min (lcms_short); [M+H + ] + =512.39.

[0236] Preparation of Compound 4.4 To a stirred solution of compound 4.3 (200.1 mg, 0.391 mmol) in DCM (3 mL) in a 20 mL glass scintillation vial was added 0.98 mL of HCl (4 M in dioxane, 10 equivalents, 3.91 mmol). As the reaction proceeded, the product precipitated from solution. Upon completion of the reaction by UPLC-MS, the reaction mixture was filtered, and the solid product was rinsed with diethyl ether (20 mL) and air-dried overnight to give crude compound 4.4 as the HCl salt. The crude compound was used in the next step without further purification. No yield was obtained from this step. UPLC-MS retention time: 2.51 min (lcms_short); [M+H + ]+ =412.32.

[0237] Preparation of Compound 4.1 To a stirred solution of methyl 4-chloro-3-hydroxy-5-nitrobenzoate (1.15 g, 5 mmol, CAS number 180031-12-3) in DMF (6 mL) in a 20 mL glass scintillation vial, 4-(3-chloropropyl)morpholine (1.1 equiv., 896 mg, 5.5 mmol, CAS number 57616-74-7) and K2CO3 (2 equiv., 1.4 g, 10 mmol) were added. The reaction mixture was brought to 70 °C and stirred overnight. After 16 h, complete conversion to compound 4.1 was observed by UPLC-MS. The reaction mixture was diluted with water (100 mL), extracted three times with methyl tert-butyl ether, and then washed three times with saturated sodium bicarbonate in water. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give compound 4.1 (1.55 g, 4.32 mmol, 86%) as an orange solid. UPLC-MS retention time: 3.08 minutes (lcms_short);[M+H + ] + =359.22.

[0238] Preparation of Compound 4.5 To a solution of compound 4.4 (202.8 mg, 0.453 mmol) in iPrOH (2 mL) in a 10 mL Discover 2.0 glass vial, compound 4.1 (1.05 equiv., 170.6 mg, 0.475 mmol) was added, followed by DIEA (0.248 mL, 1.42 mmol). The reaction vessel was sealed and then stirred at 120 °C under microwave heating (CEM Discover 2.0) for 5 h until the reaction no longer proceeded (maximum conversion 95%). The reaction mixture was diluted with water (100 mL) and extracted three times with ethyl acetate (100 mL). The combined organic phase was washed with water (100 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give crude compound 4.5 (243.5 mg) as an orange solid. Half of the product was used in the next step without purification, and the other half was purified by silica column chromatography (25G Sfar silica column) using 5→20% MeOH in DCM. Concentration of the column fractions gave compound 4.5 (61 mg, 0.0831 mmol; 18%) as an orange solid. UPLC-MS retention time: 3.32 min (lcms_short); [M+H + ] + =734.50.

[0239] Preparation of Compound 4.6 To a stirred solution of compound 4.5 (61 mg, 0.0831 mmol) in DMF (1.1 mL) in a 20 mL glass scintillation vial, NaSO (14 equiv., 202.6 mg, 1.163 mmol) was added as a solution in water (0.89 mL). After 15 min, NaHCO (43 equiv., 300 mg, 3.57 mmol) was added. After 1 h, little progress was observed, and some solids had not dissolved. 1 mL of DMF was added. An additional 200 mg of NaSO and 100 mg of NaHCO were added. The reaction mixture was stirred overnight at room temperature. After 16 h, complete conversion was observed by UPLC-MS monitoring. The reaction mixture was diluted with water (75 mL) and extracted twice with ethyl acetate (75 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (50 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give crude compound 4.6 (27 mg, 0.0384 mmol, 46%) as a yellow solid. The crude product from the organic phase was used in the next step without further purification. UPLC-MS retention time: 2.76 min (lcms_short); [M+H + ] + =704.55.

[0240] Preparation of Compound 4.7 To a stirred solution of compound 4.6 (27 mg, 0.0384 mmol) in DMF (0.5 mL) in a 4 mL glass vial at 0 °C, 23 μL of a 0.33 mg / μL solution of compound 1.11 (1 equiv., 7.49 mg, 0.0384 mmol) in dioxane was added. The compound 1.11 solution was added three times (7.6 μL each) at 15 min intervals. After UPLC-MS monitoring showed complete conversion to the thiourea intermediate, DIEA (5 equiv., 33.5 μL, 0.192 mmol) and EDC (2 equiv., 14.7 mg, 0.0768 mmol) were added, and the reaction mixture was warmed to room temperature and allowed to react overnight. After 18 h, the reaction solution changed from a clear orange solution to a cloudy yellow solution, and the reaction was deemed complete by UPLC-MS monitoring. The reaction solution was precipitated in cold diethyl ether, centrifuged, and decanted to give compound 4.7 as a yellow pellet. No yield was obtained in this step. UPLC-MS retention time: 4.87 min (lcms_long), [M+H + ] + =865.62.

[0241] Preparation of Compound 4.8 To a stirred solution of compound 4.7 (47.3 mg, 0.0547 mmol) in MeOH (0.3 mL) and THF (0.6 mL) in a 4 mL glass vial, KOH (10 equiv., 36.1 mg, 0.547 mmol) was added as a solution in 0.16 mL of water. After 2 h, an additional 19 mg of KOH in 75 μL of water was added to drive the reaction. 3 h after the second KOH addition, the reaction was judged complete by monitoring with UPLC-MS. 1 M HCl in dioxane was added to the reaction mixture until a pH of 3 was achieved (1 mL). The reaction mixture was diluted with water, frozen, and lyophilized. The resulting solid was resuspended in 90% DCM, 9% MeOH, 1% DIEA, and the solid was removed by filtration. The filtrate was concentrated in vacuo to give compound 4.8 (40.5 mg, 0.0476 mmol, 87%) as a solid. UPLC-MS retention time: 4.06 min (lcms_long), [M+H + ] + =851.58.

[0242] Preparation of Compound 4.9 To a stirred solution of compound 4.8 (40.5 mg, 0.0476 mmol) in DCM (0.6 mL) in a 4 mL glass vial, DIEA (6 equiv., 49.7 μL, 0.2855 mmol), HOSu (1.4 equiv., 7.66 mg, 0.0666 mmol), and DIC (1.4 equiv., 10.3 μL, 0.0666 mmol) were added. The reaction mixture was brought to 40 °C and stirred for 8 h until UPLC-MS monitoring indicated maximum conversion (the reaction stopped after 95% conversion). The reaction mixture was concentrated in vacuo to give compound 4.9 (42.8 mg, 0.0451 mmol, 95%) as a solid. The product was used in the next step without further purification. UPLC-MS retention time: 4.33 min (lcms_long), [M+H + ] + =948.53.

[0243] Preparation of Compound III To a stirred solution of compound 4.9 (30 mg, 0.03168 mmol) in MeCN (2 mL) in a 20 mL glass scintillation vial, hydrazine monohydrate (3 equiv., 7.32 mg, 0.095 mmol) was added. After 5 min, all of the starting material was consumed, as observed by UPLC-MS monitoring. The reaction mixture was diluted with 1 mL of 1:1 MeCN:HO and purified by RP-HPLC (Phenomenex Luna C5 250 x 21.2 mm column, 20 mL / min flow rate, 0.05% TFA, 15-30% MeCN / HO gradient over 35 min, 50 min total run time). Pure fractions were combined and lyophilized to afford compound III (6.2 mg, 0.00717 mmol, 23%) as a white powder. UPLC-MS retention time: 3.46 min (lcms_long_C4), [M+H + ] + =865.59. [Example]

[0244] Synthetic Scheme IV [ka]

[0245] Preparation of tert-butyl 4-({2-amino-1-[(2E)-4-(2-amino-5-carbamoyl-7-methoxy-1H-1,3-benzodiazol-1-yl)but-2-en-1-yl]-5-carbamoyl-1H-1,3-benzodiazol-7-yl}oxy)butanoate (5.1) To a solution of compound 1.8 (1.58 g, 2.90 mmol, 1.0 equiv.) in N-methyl-2-pyrrolidone (25.2 mL) was added cyanogen bromide (770 mg, 7.26 mmol, 2.5 equiv.) at room temperature. The reaction mixture was stirred at 60° C. for 15 hours, cooled to room temperature, and diluted with diethyl ether (75 mL). The mixture was filtered, and the cake was washed with diethyl ether (2×25 mL). The solid was collected by filtration to give a beige solid (3.41 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 120 g) using dichloromethane / (methanol / ammonium hydroxide 9 / 1) (95 / 5 for 5 min, then 70 / 30 over 30 min, 70 / 30 for 15 min) as the eluent to give impure compound 5.1 (1.31 g) as a beige solid after coevaporation with acetonitrile (3 × 50 mL). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 40 g) using dichloromethane / (methanol / ammonium hydroxide 9 / 1) (90 / 10 to 70 / 30 over 30 min, 70 / 30 for 20 min) as the eluent to give impure compound 5.1 (1.30 g) as a beige solid after coevaporation with acetonitrile (3 × 50 mL). The residue was suspended in methanol (100 mL) and activated Amberlyst A26-OH- (3.0 g) was added. The suspension was rotary evaporated at room temperature for 15 hours. Water (100 mL) was added to the mixture and rotary evaporated for 30 minutes. The suspension was filtered through cotton, and the filtrate was concentrated to dryness. After co-evaporation with acetonitrile (3 × 50 mL), pure compound 5.1 (851 mg, 1.44 mmol, 49%) was obtained as a beige solid. 1H NMR (DMSO-d6): δ1.38(s,9H), 1.82~1.88(m,2H), 2.31(t,J=7.2Hz,2H), 3.69(s,3H), 3.96(t,J=6.3Hz,2H), 4.73 (dd,J=4.8,11.7Hz,4H), 5.58~5.71(m,2H), 6.36(d,J=10.5Hz,4H), 7.04(d,J=3.8Hz,4H), 7.37(s,2H), 7.77(br s,2H). LCMS (2~100 ACN / H2O+0.1%AF): Tr=6.28 min; Purity=99.57%; [M+H + ] + =593.18.

[0246] Compound 5.1 was obtained pure after flash chromatography alone in 63% yield.

[0247] Preparation of Compound 5.2 To a solution of compound 5.1 (150 mg, 0.25 mmol, 1.0 equiv) in DMSO (3.0 mL) was added 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (CAS#1564709-36-9, 98.1 mg, 0.63 mmol, 2.5 equiv), HATU (240 mg, 0.63 mmol, 2.5 equiv), and DIEA (348.6 μL, 2.0 mmol, 8 equiv). The reaction mixture was stirred at 105 °C for 6 h, cooled to room temperature, and triturated with cold diethyl ether (75 mL). The precipitate formed was isolated by centrifugation. The residue was purified by flash chromatography (5→20% MeOH / DCM) on silica gel (60 μm amorphous, 25 g) to give compound 5.2 as a beige solid (95 mg, 0.11 mmol, 44%). UPLC-MS retention time: 5.08 min (lcms_long), [M+H + ] + =867.54.

[0248] Preparation of compound 5.3 To a suspension of compound 5.2 (95 mg, 0.11 mmol) in THF (4 mL) at room temperature, HCl (4 M in dioxane, 4.0 mL, 16.0 mmol) was added, and the resulting mixture was stirred at room temperature for 24 hours. Complete conversion to compound 5.3 was confirmed by monitoring with UPLC-MS. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the resulting precipitate was isolated by centrifugation. The supernatant was removed, and the resulting pellet was resuspended in cold diethyl ether (100 mL) and centrifuged. This step was repeated twice. Finally, the pellet was dried in vacuo to give compound 5.3 as the HCl salt, a beige solid (80 mg; 0.098 mmol, 90%). The product was used in the next step without further purification. UPLC-MS retention time: 3.82 min (lcms_long), [M+H + ] + =811.53.

[0249] Preparation of Compound 5.4 To a suspension of compound 5.3 (20 mg, 0.024 mmol, 1.0 equiv., HCl salt) in DMF (1 mL) at 40 °C, DIEA (12.9 μL, 0.074 mmol, 3.0 equiv.) was added, followed by the addition of N-hydroxysuccinimide (13.8 mg, 0.12 mmol, 5.0 equiv.) and N,N'-diisopropylcarbodiimide (18.4 mg, 0.144 mmol, 6.0 equiv.). The resulting mixture was monitored by UPLC-MS until the reaction no longer proceeded (maximum conversion of 25%). The reaction mixture was added dropwise to cold diethyl ether (40 mL), and the precipitate was isolated by centrifugation. The pellet was dried in vacuo to give compound 5.4 as a beige solid. The crude product (21 mg, a mixture of 25% compound 5.4 and 75% compound 5.3) was used in the next step without further purification. UPLC-MS retention time: 4.1 min (lcms_long), [M+H + ] + =908.55.

[0250] Preparation of Compound IV Compound IV was prepared according to general procedure C using N,N'-dimethylhydrazine dihydrochloride to give a beige solid. UPLC-MS retention time: 3.69 min (1cms_long), [M+H + ] + =853.51. [Example]

[0251] The following compound V was prepared using the general methods disclosed herein. [ka] [Example]

[0252] The following compound VI was prepared using the general methods disclosed herein. [ka] [Example]

[0253] The following compound VII was prepared using the general methods disclosed herein. [ka] [Example]

[0254] The following compound VIII was prepared using the general methods disclosed herein. [ka] [Example]

[0255] The following compound IX was prepared using the general methods disclosed herein. [ka] [Example]

[0256] The following compounds X were prepared using the general methods disclosed herein. [ka] [Example]

[0257] The following compounds XI were prepared using the general methods disclosed herein. [ka] [Example]

[0258] The following compounds XII were prepared using the general methods disclosed herein. [ka] [Example]

[0259] The following compounds XIII were prepared using the general methods disclosed herein. [ka] [Example]

[0260] The following compound XIV was prepared using the general methods disclosed herein. [ka] [Example]

[0261] The following compound XV was prepared using the general methods disclosed herein. [ka] [Example]

[0262] The following compound XVI was prepared using the general methods disclosed herein. [ka] [Example]

[0263] The following compound XVII was prepared using the general methods disclosed herein. [ka] [Example]

[0264] The following compound XVIII was prepared using the general methods disclosed herein. [ka] [Example]

[0265] The following compound XIX was prepared using the general methods disclosed herein. [ka] [Example]

[0266] THP-1 assay THP1 Dual Cells™ were purchased from Invivogen (catalog number thpd-nfis) and used to determine the biological activity of compounds at the molecular level and to determine downstream activation of STING signaling (IRF and NFκB pathway activation). Briefly, cultured cells were plated at 1 × 10 per well on the day of the experiment. 5Cells were seeded at a density of 1000 x g (96-well format in RPMI supplemented with 25 mM HEPES, 10% heat-inactivated FCS). Serial dilutions of compounds were prepared and added to the cells, which were then incubated for a total of 24 hours. Test articles were tested at eight concentrations to determine dose-dependent activation of the NFκB and IRF pathways. After 24 hours, cells were centrifuged, and supernatants were collected for detection of secreted alkaline phosphatase (QUANTI-Blue™ detection solution, catalog number rep-qbs, measured by Invivogen) or lucia luciferase (QUANTI-Luc™ reagent, catalog number rep-qlc1, measured by Invivogen), respectively. Readouts were performed according to the manufacturer's instructions. EC50 values ​​were calculated using GraphPad Prism 9. THP-1 plots for compounds IA and ID are shown in Figures 1A and 1B. [Example]

[0267] HEK STING assay with PFO HEK293 cells with knock-in STING receptors were purchased from Invivogen (catalog number 293dh232) and used to determine the biological activity of compounds with and without the cell membrane permeabilizing agent PFO. Activity was determined by analyzing downstream activation of STING signaling (IRF and IFNβ pathway activation). Briefly, cells were cultured at 2 × 10 per well. 4Cells were seeded (in DMEM in a 96-well format) and incubated overnight. The next day, serial dilutions of the components were prepared and added to the cells for a total of 24 hours of incubation. Test articles were tested at eight concentrations to determine dose-dependent activation of the IRF pathway and IFN-β secretion. In addition to the test article, 50 ng / ml of PFO was added to the wells. After 24 hours of incubation with the test article with or without PFO (US Biological Life Sciences, Cat. No. 370743), the cells were centrifuged, and the supernatants were collected for detection of secreted alkaline phosphatase (measured by QUANTI-Blue™ detection solution, Cat. No. rep-qbs, Invivogen) or lucia luciferase (measured by QUANTI-Luc™ reagent, Cat. No. rep-qlc1, Invivogen), respectively. Readout was performed according to the manufacturer's instructions. [Example]

[0268] STING haplotype assay THP1 Dual Cell™ cells carrying different human STING variants were purchased from Invivogen: THP-1Dual™ KI-hSTING-H232 cells (catalog no. Thpd-h232) and THP-1Dual™ KI-hSTING-R232 cells (catalog no. thpd-r232). In these cell lines, naturally expressed STING was first knocked out, followed by knocking in the coding sequence of the human STING variant. R232 is described by Invivogen as occurring in approximately 45-58% of the human population, while h232 occurs in approximately 14%. The biological activity of the compounds was evaluated as described in Example 20. The STING haplotype profiles of compounds IA and ID are shown in Figures 2A-2D. [Example]

[0269] PBMC assay Compounds IA-IV were further tested in PBMCs from healthy donors to determine dose-dependent cytokine induction. Buffy coat-derived PBMCs were thawed on the day of the experiment and 5x10 per well. 5 Cells were seeded at a density of 1000 x g (96-well format, RPMI containing 5% human serum, 1% MEM non-essential amino acids, and 1% sodium pyruvate). Serial dilutions of compounds were prepared as described in the previous experiment and added to the cells for 24 hours. The next day, cells were centrifuged, and the supernatant was collected for cytokine readout based on a multiplex assay kit from mesoscale discovery (customized multiplex panel for cytokines: IFNα2a, IFNβ, TNFα, and IL6). The effect on cell viability was determined by Promega's CellTiter Glo 2.0 (Cat. No. G9241). Readouts for both experiments were performed according to the manufacturer's instructions. Representative curves for PBMCs for compounds IA and ID are shown in Figures 3A-5B. [Example]

[0270] STING production in humans and mice T7-shuffled E. coli cells transformed with expression vectors encoding either human or mouse His-tagged truncated versions of STING were cultured overnight in 50 mL of LB medium. 200 mL of LB medium was inoculated to an OD of 0.05, and the cells were grown at 37°C and 120 rpm until the OD reached 0.5. Protein expression was induced using 1 mM IPTG, and the cells were further cultured overnight at 30°C. Cells were harvested by centrifugation at 4000 rpm for 30 minutes at 4°C and frozen at -80°C. Cells were then thawed and resuspended in 30 mL of IMAC-buffer A (10 mM HEPES, 50 mM NaCl, 10 mM imidazole, 250 mM L-arginine, 10% glycerol, pH 7.1) supplemented with 15 U / mL benzonase, 0.2 mg / mL MgCl2, and 0.1 mg / mL lysozyme. Lysis was performed using five sonication cycles with 25% amplitude and 2-second pulses, with a 5-minute incubation on ice between cycles. The lysate was centrifuged at 16,000 g for 15 minutes, and the supernatant was filtered (0.22 μm). STING purification was performed using an AKTA Prime chromatography system. A 1 mL HisTrap Excel column was equilibrated with IMAC-Buffer A and then loaded with the lysate. After washing with multiple column volumes of buffer, STING was eluted using a linear gradient of IMAC-Buffer B (10 mM HEPES, 50 mM NaCl, 500 mM imidazole, 250 mM L-arginine, 10% glycerol, pH 7.1). Elution fractions were analyzed by SDS-PAGE, and STING-containing fractions were combined. STING was further purified by preparative size-exclusion chromatography using an AKTA Pure system coupled with a HiLoad Superdex 16 / 600 75 pg column. Fractions were analyzed by SDS-PAGE and STING-containing fractions were combined.

[0271] The purified protein was biotinylated using a 10-fold excess of EZ-Link Sulfo-NHS-LC-Biotin (Life Technologies) for 2 hours on ice. The reaction was quenched by adding 100 mM Tris, pH 8, and incubating on ice for 1 hour. Residual biotin was removed by buffer exchange into PBS using Zeba Spin Desalting Columns, 7K MWCO, 0.5 mL (Thermo Fischer). Protein concentration was determined using a Nanodrop system. [Example]

[0272] Affinity measurement assay Affinity measurements of STING-binding compounds were performed using a Biacore T200 SPR system (GE Healthcare, Biacore T200 control Software 3.2). Therefore, the Biotin CAPture kit, series S (Cytiva Europe, catalog no. 28920234) was utilized. The newly docked Biotin CAPture chip was rehydrated overnight in the instrument (standby mode). The following day, the chip surface was conditioned three times for 60 seconds with regeneration solution (6 M guanidine-HCl, 0.25 M NaOH) at 10 μL / min on flow cells (FC) 1 and FC2. Subsequently, Biotin CAPture reagent (Cytiva Europe, catalog no. 29423383) was delivered to FC1 and FC2 at 2 μL / min for 300 seconds, followed by capture of 100 nM biotinylated mouse or 150 nM biotinylated human STING, respectively. STING immobilization was performed on FC2 for 600 seconds at 10 μL / min, followed by quenching of unbound streptavidin with 0.1 mM biocytin for 60 seconds on both flow cells and at 30 μL / min. Five injections of STING-binding compounds at 10 nM, 5 nM, 2.5 nM, 1.25 nM, 0.625 nM, and 0 nM (blank control) were performed on both flow cells for 120 seconds at a flow rate of 30 μL / min, followed by a 900-second dissociation period. After each cycle, the surface was regenerated with regeneration solution for 120 seconds at 10 μL / min. All measurements were performed at 25°C using HBS-EP+ buffer (Cytiva Europe, catalog no. BR100669). The control response from FC1 was subtracted from the sample measurement on FC2 (FC2-FC1), followed by subtraction of the 0 nM blank injection. The resulting binding curves of this multi-cycle kinetic experiment were fitted using Biacore T200 Evaluation Software 3.2 and a 1:1 binding model. [Example]

[0273] Table of activity and binding affinity of compounds IA-IV The activities and binding affinities of compounds IA to IV are shown in Tables 2 to 4 below.

[0274] [Table 7] JPEG2026508759000162.jpg42170

[0275] [Table 8] JPEG2026508759000164.jpg157170

[0276] [Table 9] JPEG2026508759000166.jpg33170 [Example]

[0277] Antitumor effect of the comparative compound diABZI in 4T1 Antitumor effect of the comparative compound diABZI (catalog number 2138299-33-7) having the following structure: [ka]

[0278] was tested in a murine 4T1 breast cancer model. Balb / c RJ mice were injected with 1 × 10 5 4T1 cells were injected subcutaneously. Three doses of agonist diluted in PBS (31 nmol, 3.1 nmol and 0.31 nmol) were administered on days 11, 16 and 21 post-inoculation and compared with an untreated control group. Tumor size was monitored using caliper measurements up to day 21 (volumes were On day 21, animals were sacrificed and tumor cytokines were analyzed relative to serum 3 hours after the final agonist injection. To assess the statistical significance of tumor volume reduction compared to untreated, a two-way ANOVA test was used:*** p<0.0001.

[0279] Tumor growth data for diABZI are shown in Figures 9A-9E. Cytokine data for diABZI are shown in Figures 7B and 8B. [Example]

[0280] Antitumor effect of compound ID in 4T1 The antitumor effect of compound ID was tested in a mouse 4T1 breast cancer model. Balb / c Rj mice were treated with 1 × 10 5 4T1 cells were subcutaneously injected. Three doses of Compound ID (31 nmol, 3.1 nmol, and 0.31 nmol) formulated in 5% Solutol in 2.5% glucose or vehicle control (5% Solutol in 2.5% glucose) were administered intratumorally on days 11, 16, and 21 post-inoculation. Tumor size was monitored using BioVolume® (tumor size calculated based on RGB, thermal, and 3D model data) until day 21, at which time animals were sacrificed and tumor cytokines were analyzed relative to serum 3 hours after the final agonist injection. A two-way ANOVA test was used to assess the statistical significance of tumor volume reduction compared to vehicle control: *** p<0.0001.

[0281] Tumor growth data for each treatment group, measured using BioVolume, are shown in Figures 6A-6E. As can be seen from these figures, administration of Compound ID at various doses resulted in a significant reduction in tumor size when compared to vehicle controls.

[0282] The concentrations of IFNα and IFNγ in the tumors for each treatment group and the untreated control are shown in Figures 7A and 8A. As shown in Figures 7A-8B, compound ID induced higher levels of IFNα and IFNγ cytokines in the tumors, indicating better activation of the immune system in the tumors when compared to the comparative compound diABZI. [Example]

[0283] Synthesis scheme of compound XX [ka]

[0284] Preparation of (3-bromopropoxy)tris(propan-2-yl)silane (23-2) [ka]

[0285] In a flask under argon, triisopropylsilyl chloride (76.9 mL, 360 mmol, 1.00 equiv.) followed by imidazole (61.2 g, 899 mmol, 2.50 equiv.) was added to a solution of 3-bromopropan-1-ol (50.0 g, 360 mmol, 1.00 equiv.) in dichloromethane (360 mL). The reaction mixture was stirred at room temperature for 2.5 days. Dichloromethane (75 mL) and water (75 mL) were added to the reaction mixture. The reaction mixture was diluted with water (500 mL). The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 500 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give an opaque oil (109 g). The residue was purified by silica gel chromatography (silica: 970 g, liquid loading in cyclohexane) using cyclohexane as the eluent to give pure compound 23-2 (80.3 g, 272 mmol, 76%) as a colorless oil.

[0286] NOTE: The product is only detected by TLC (cyclohexane / ethyl acetate 9:1, stain: potassium permanganate).

[0287] 1 H NMR (CDCl3): δ1.02~1.32(s,21H), 2.04(tt,2H,J=6.0,6.0Hz), 3.53(t,2H,J=6.0Hz), 3.80(t,2H,J=6.0Hz).

[0288] Preparation of 4-chloro-3-nitro-5-(3-{[tris(propan-2-yl)silyl]oxy}propoxy)benzamide (23-3) [ka]

[0289] In a flask under argon, a suspension of compound 4-chloro-3-hydroxy-5-nitrobenzamide 10-5 (10.6 g, 46.2 mmol, 1.00 equiv.), compound 23-2 (16.4 g, 55.4 mmol, 1.20 equiv.), and anhydrous potassium carbonate (8.30 g, 60.0 mmol, 1.30 equiv.) in anhydrous N,N-dimethylformamide (59 mL) was stirred at 70 °C for 18 h. The reaction mixture was concentrated in vacuo. The residue was suspended in dichloromethane (200 mL) and sonicated at room temperature for 20 min. The solid was collected by filtration to give a beige solid (25.4 g). The residue was purified by silica gel flash chromatography in two portions of 12.7 g each (330 g, 50 μm amorphous, solid loading on silica) using dichloromethane / methanol (100:0 for 5 min, 97.5:2.5 over 30 min, and 97.5:2.5 for 10 min) as eluent to give compound 23-3 (20.4 g, 45.2 mmol, 98%) as a pale yellow solid contaminated with 4.64 wt% N,N-dimethylformamide.

[0290] 1 H NMR (DMSO-d6): δ0.95~1.36(m,21H), 2.00(dd,2H,J=5.9,5.9Hz), 3.88(d,2H,J=5.9Hz), 4. 33(d,2H,J=5.9Hz), 7.76(s,1H), 7.88(d,1H,J=1.8Hz), 8.04(d,1H,J=1.8Hz), 8.28(s,1H).

[0291] LCMS (2-100 ACN / H2O+0.1%FA, 5 min): Tr=3.85 min, purity=100%, [M+H + ]=443.2.

[0292] Preparation of tert-butyl N-[(2E)-4-{[4-carbamoyl-2-nitro-6-(3-{[tris(propan-2-yl)silyl]oxy}propoxy)phenyl]amino}but-2-en-1-yl]carbamate (36-1) [ka]

[0293] In an argon-purged sealed flask, N,N-diisopropylethylamine (4.00 equiv., 16.9 mL, 96.9 mmol) was added to a stirred solution of compound 23-3 (1.00 equiv., 12.0 g, 24.2 mmol) and tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (1.50 equiv., 6.77 g, 36.3 mmol) in isopropanol (80 mL) at room temperature. The flask was sealed, and the reaction mixture was stirred at 125 °C for 18 hours. The reaction mixture was diluted with water (400 mL) and ethyl acetate (400 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product (16.5 g) as an orange oil. The residue was purified by silica gel flash chromatography (column: Interchim 15 μm 330 g, loading: solid (silica, 45 g)) using cyclohexane / ethyl acetate (gradient from 100:00 to 70:30 over 30 min, then 70:30 for 45 min, then 20:80 over 25 min) as the eluent to give the desired compound 36-1 (11.2 g, 19.1 mmol, 79%, contaminated with 1.0 wt% ethyl acetate) as a red solid.

[0294] LC / MS (CSH, water+0.1%HCO2H / MeCN 2~100 gradient, 3.5 min): Rt=2.04 min, 100%, [M-tBu+H] + =525.85

[0295] 1H NMR(DMSO-d6,400MHz):δppm 8.19(d,J=1.8Hz,1H), 8.01(s,1H), 7.73(t,J=6.1Hz,1H), 7.56(d,J=2.0Hz,1H), 7.29(s,1H), 6.91(t,J=6.0Hz,1H), 5.63~5 .48(m,2H), 4.24~4.08(m,4H), 3.86(t,J=6.2Hz,2H), 3.55~3.41(m,2H), 2.07~1.93(m,2H), 1.35(s,9H), 1.16~0.96(m,21H).

[0296] Preparation of tert-butyl N-[(2E)-4-{[2-amino-4-carbamoyl-6-(3-{[tris(propan-2-yl)silyl]oxy}propoxy)phenyl]amino}but-2-en-1-yl]carbamate (36-2) [ka]

[0297] In a flask, compound 36-1 (1.00 equiv., 8.82 g, 15.2 mmol) was suspended in methanol (23 mL), N,N-dimethylformamide (15 mL), and water (35 mL) at room temperature. A solution of sodium dithionite (6.00 equiv., 15.9 g, 91.1 mmol) in water (55 mL) was added at room temperature, and the suspension was stirred for 5 minutes. Sodium bicarbonate (12.0 equiv., 15.3 g, 182 mmol) was then added. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water (200 mL) and ethyl acetate (200 mL). The aqueous layer was extracted with ethyl acetate (2 × 150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product (12.4 g) as a yellow oil. The crude product was purified by silica gel flash chromatography (column: Interchim 50 μm 330 g, loading: solid (silica, 36 g)) using dichloromethane / methanol (10:0 for 5 min, then gradient to 95:5 over 25 min, then 95:5 for 25 min) as the eluent to give the desired compound 36-2 (6.17 g, 9.89 mmol, 65%, contaminated with 11.7 wt% dichloromethane) as an off-white solid.

[0298] NOTE: If the reaction is not complete, more sodium dithionite in water can be added to reach completion (indicated by disappearance of the orange color in a deep white or very pale yellow solution).

[0299] LC / MS (CSH, water + 0.1% HCO2H / MeCN 2~100 gradient, 3.5 min): Rt=1.90 min, 97%, [M+H] + =552.36)

[0300] 1H NMR(DMSO-d6,400MHz):δppm 7.58(s,1H), 7.00~6.44(m,4H), 5.69~5.44(m,2H), 4.65(s,2H), 4.04(t,J=6.0Hz,2H), 3.85(t,J=6.3Hz,2H) , 3.74(d,J=8.2Hz,1H), 3.52(dd,J=13.2,6.3Hz,4H), 1.94(p,J=6.2Hz,2H), 1.37(s,9H), 1.18~0.91(m,21H). [ka]

[0301] Preparation of 4-ethyl-2-methyl-1,3-oxazole-5-carbonyl chloride (29-3) [ka]

[0302] In a flask under argon, oxalyl chloride (1.20 equiv., 4.98 mL, 58.0 mmol) was added to a suspension of compound 29-1 (1.00 equiv., 7.50 g, 48.3 mmol) in anhydrous dichloromethane (250 mL) at room temperature (gentle bubbling occurred). Anhydrous N,N-dimethylformamide (0.10 equiv., 0.37 mL, 4.83 mmol) was then added dropwise (vigorous bubbling occurred), and the solution was stirred at room temperature for 18 h (the white suspension turned into a pale yellow solution and bubbling ceased). All volatiles (except N,N-dimethylformamide) were removed under vacuum, and the crude product (8.39 g, 48.3 mmol, 100%) was used in the next step without further purification.

[0303] LC / MS samples were prepared by diluting an aliquot of the reaction into a vial of anhydrous methanol and then heating for a few seconds.

[0304] LC / MS (CSH, water + 0.1% HCO2H / MeCN 2~100 gradient, 3.5 min): Rt=1.24 min, 100%, [M+H] + = 170.13 (as methyl ester of 29-3)

[0305] Preparation of 4-ethyl-2-methyl-1,3-oxazole-5-carbonyl isothiocyanate (29-2) [ka]

[0306] In a flask under argon, potassium thiocyanate (1.30 equiv., 6.11 g, 62.8 mmol) was suspended in acetone (70 mL) and cooled to 0 °C. Then, a solution of compound 29-3 (1.00 equiv., 8.39 g, 48.3 mmol) in acetone (145 mL) was added dropwise at 0 °C. The reaction mixture was stirred at 0 °C for 3 h. The reaction mixture was concentrated in vacuo and coevaporated with n-hexane (3 × 50 mL). The residue was suspended in n-hexane (100 mL), filtered, washed with n-hexane (3 × 150 mL), and the filtrate was concentrated in vacuo to give the crude product (8.75 g) as a yellow oil. The crude product was purified by silica gel flash chromatography (column: Interchim 50 μm 330 g, loading: solid (Celite, 24 g)) using cyclohexane / ethyl acetate (gradient 10:0 to 7:3 over 30 min) as the eluent to give the desired compound 29-2 (8.26 g, 40.9 mmol, 85%, contaminated with 2.9 wt% dichloromethane) as a pale yellow oil.

[0307] 1 H NMR (CDCl3,400MHz): δppm 2.89 (q, J = 7.6 Hz, 2H), 2.53 (s, 3H), 1.26 (t, J = 7.6Hz, 3H).

[0308] Preparation of tert-butyl N-[(2Z)-4-[5-carbamoyl-2-(4-ethyl-2-methyl-1,3-oxazol-5-amido)-7-(3-{[tris(propan-2-yl)silyl]oxy}propoxy)-1H-1,3-benzodiazol-1-yl]but-2-en-1-yl]carbamate (36) [ka]

[0309] In a flask under argon, a solution of compound 36-2 (1.00 equiv., 5.40 g, 9.80 mmol) in N,N-dimethylformamide (95 mL) was cooled to 0 °C. Then, a solution of compound 29-2 (0.2 M in dioxane, 1.00 equiv., 49.0 mL, 9.80 mmol) was added dropwise via a dropping funnel over 45 minutes. The reaction mixture was then stirred at 0 °C for 1 hour. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.00 equiv., 3.76 g, 19.60 mmol) and triethylamine (4.00 equiv., 3.97 g, 5.45 mL, 39.2 mmol) were then added at 0 °C. The reaction mixture was allowed to warm to room temperature and stirred for 18 hours. The reaction mixture was diluted with dichloromethane (300 mL) and water / saturated aqueous ammonium chloride solution (3:1, 200 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product (10.1 g) as a waxy beige solid. The crude product was purified by silica gel flash chromatography (column: Interchim 15 μm 330 g, loading: solid (silica, 30 g)) using dichloromethane / methanol (100:0 for 5 min, then gradient to 90:10 over 45 min) as the eluent to give the desired compound 36 (6.40 g, 8.72 mmol, 89%, contaminated with 2.8 wt% dichloromethane) as a pale yellow foamy solid.

[0310] Note: If the cyclization step is not complete, additional N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and triethylamine can be added to achieve complete conversion of the intermediate to the desired product. The reaction is robust and can be stirred at room temperature for several days, if necessary.

[0311] LC / MS (CSH, water + 0.1% HCO2H / MeCN 50-100 gradient, 12.0 min): Rt=6.59 min, 98.00%, [M+H] + =714.03

[0312] 1 H NMR(DMSO-d6,400MHz):δppm 12.73(s,1H), 7.96(s,1H), 7.65(d,J=1.3Hz,1H), 7.46~7.16(m,2H), 6.87(s,1H) ), 5.78~5.66(m,1H), 5.64~5.50(m,1H), 4.91(d,J=5.8Hz,2H), 4.27(t,J=6.1Hz, 2H), 3.89(t,J=6.2Hz,2H), 3.48(s,2H), 2.99(q,J=7.5Hz,2H), 2.44(s,3H), 2.0 5(tt,J=6.2,6.1Hz,2H), 1.31(s,8H), 1.19(t,J=7.5Hz,4H), 1.10~0.99(m,21H).

[0313] Preparation of 36-A [ka]

[0314] To a solution of compound 36 (1.08 g, 1.51 mmol, 1.0 equiv.) in dichloromethane (20.0 mL) was added dropwise 4 M HCl in dioxane (3 mL, 12.1 mmol, 8 equiv.). The resulting solution was stirred at room temperature for 30 min. Complete removal of the Boc and TIPS protecting groups was confirmed by UPLC-MS analysis. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the formed precipitate was isolated by centrifugation. The solid was rinsed with diethyl ether (50 mL) and dried under reduced pressure to give 36-A (quantitative) as an off-white solid.

[0315] [M-HCl + H + ] + =457.3.

[0316] Preparation of 36-B [ka]

[0317] To a suspension of compound 36-A (2.1 g, 3.98 mmol, 1.0 equiv.) in acetonitrile (10.0 mL) was added methyl 6-chloro-5-nitronicotinate (1.0 g, 4.78 mmol, 1.2 equiv.) as a solid, followed by N,N-diisopropylethylamine (3.0 equiv., 2.1 mL, 12 mmol). The resulting suspension was stirred at 45 °C for 1 h. High-purity conversion to the desired product was confirmed by UPLC-MS analysis. The reaction mixture was added dropwise to cold diethyl ether (100 mL), and the formed precipitate was isolated by centrifugation. The solid was rinsed with diethyl ether (50 mL) and dried under reduced pressure to give crude 36-B as a yellow material, which was used directly in the next step without further purification.

[0318] [M+H + ] + =637.3.

[0319] Preparation of 36-C [ka]

[0320] To a solution of compound 36-B (approximately 3.90 mmol, 1.0 equiv.) in acetonitrile, dichloromethane, and N,N-dimethylformamide (30.0 mL, 10 mL, and 10 mL, respectively), triisopropylsilyl chloride (5.3 g, 27.3 mmol, 7.0 equiv.) was added, followed by imidazole (2.5 equiv., 0.68 g, 10.0 mmol) and N,N-diisopropylethylamine (5.9 equiv., 4 mL, 22.9 mmol). The resulting solution was stirred at 45 °C for 1 h. Conversion to the desired product was confirmed by UPLC-MS analysis. The volatiles were removed under reduced pressure, and the residue was diluted with water (200 mL) and extracted with ethyl acetate (2 × 150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product (1.56 g, 1.9 mmol) as a yellow solid, which was used directly in the next step without further purification.

[0321] [M+H +] + =793.5.

[0322] Preparation of 36-D [ka]

[0323] In a 100 mL round-bottom flask, compound 36-C (1.00 equiv., 1.56 g, 1.9 mmol) was suspended in methanol (10 mL), N,N-dimethylformamide (20 mL), and water (10 mL) at room temperature. A solution of sodium dithionite (30.0 equiv., 10.0 g, 57.4 mmol) in water (20 mL) was added at room temperature, and the resulting heterogeneous mixture was stirred for 10 minutes. Sodium bicarbonate (62.0 equiv., 10.0 g, 118.9 mmol) was then added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (200 mL) and ethyl acetate (200 mL). The aqueous layer was extracted with ethyl acetate (2 × 150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was added dropwise to cold diethyl ether (100 mL) to give the crude product. After purification by silica column chromatography (25G Sfar silica column) using 5→15% MeOH in DCM, the product was isolated as a beige solid (1.1 g, 1.38 mmol, 73%).

[0324] [M+H + ] + =763.6.

[0325] Preparation of 36-E [ka]

[0326] To a solution of compound 36-D (0.44 g, 0.57 mmol, 1.0 equiv.) in methanol (12 mL) was added cyanogen bromide (10.0 equiv., 0.611 g, 5.77 mmol), followed by N,N-diisopropylethylamine (1.0 equiv., 0.1 mL, 0.57 mmol). The resulting solution was stirred at 40 °C for 16 h. Conversion to the product without the TIPS protecting group was confirmed by UPLC-MS analysis. The volatiles were removed under reduced pressure, and the residue was added dropwise to cold diethyl ether (100 mL). The precipitate that formed was dried under reduced pressure to give the crude product (0.4 g, approximately 75% pure by UPLC-MS) as a brownish solid. The crude product was used in the next step without further purification.

[0327] [M+H + ] + =632.4.

[0328] Preparation of 36-F [ka]

[0329] To a solution of compound 36-E (0.3 g, 0.47 mmol, 1.0 equiv.) in N,N-dimethylformamide (4 mL) was added 4-ethyl-2-methyloxazole-5-carboxylic acid (2.7 equiv., 196.5 mg, 1.26 mmol), HATU (2.7 equiv., 481.2 mg, 1.26 mmol), and N,N-diisopropylethylamine (5.3 equiv., 0.44 mL, 2.51 mmol). The resulting solution was stirred at room temperature for 16 h. Complete conversion to the product was confirmed by UPLC-MS analysis. The reaction mixture was added dropwise to cold diethyl ether (25 mL), and the formed precipitate was purified by silica column chromatography (10G Sfar silica column) using 5→10% MeOH in DCM to give a beige solid (205 mg, approximately 75% pure by UPLC-MS).

[0330] [M+H + ] + =906.4.

[0331] Preparation of 36-G [ka]

[0332] To a solution of compound 36-F (153.7 mg, 0.17 mmol, 1.0 equiv.) in tetrahydrofuran, methanol, and water (1 mL each) was added 5 M aqueous NaOH (5.9 equiv., 0.2 mL, 1 mmol). The resulting solution was stirred at 40 °C for 1 h. Complete conversion to the product was confirmed by UPLC-MS analysis, and the volatiles were removed under reduced pressure. The residue was diluted with water (5 mL), and the pH was adjusted to 6 by adding 1 M aqueous HCl. The mixture was then lyophilized to give a beige solid, which was used in the next step without further purification.

[0333] [M+H + ] + =755.4.

[0334] Preparation of 36-H [ka]

[0335] To a solution of compound 36-G (approximately 0.17 mmol, 1.0 equiv.) in N,N-dimethylformamide (8 mL) was added ammonium chloride (10 equiv., 90.9 mg, 1.70 mmol), HATU (2.5 equiv., 161.6 mg, 0.42 mmol), and N,N-diisopropylethylamine (12.0 equiv., 0.35 mL, 2.04 mmol). The resulting solution was stirred at room temperature for 5 minutes. Complete conversion to the product was confirmed by UPLC-MS analysis. The volatiles were removed under reduced pressure, and the residue was purified by silica column chromatography (10G Sfar silica column) using 5→25% MeOH in DCM to give a beige solid (86 mg, 0.11 mmol, 67%).

[0336] [M+H + ] + =754.5.

[0337] Preparation of Compound XX [ka]

[0338] To a solution of 36-H (1.00 equiv., 76 mg, 0.1 mmol) in N-methyl-2-pyrrolidone (0.7 mL) at 0 °C was added triethylamine (9 equiv., 123.1 μL, 0.9 mmol), followed by methanesulfonyl chloride (7 equiv., 54.1 μL, 0.7 mmol). After 1 h, monitoring the reaction by UPLC-MS showed 90% conversion to the mesylate. The reaction mixture was added dropwise to cold diethyl ether (20 mL), and the precipitate that formed was collected by centrifugation. To the solid was added N-methyl-2-pyrrolidone (0.7 mL) and methylhydrazine (38.2 equiv., 200 μL, 3.8 mmol). The resulting solution was stirred at 45 °C for 1 h and was complete as shown by UPLC-MS. Purification was carried out using RP-HPLC to afford the title compound as a trifluoroacetate salt (15 mg, 0.017 mmol, 17% over two steps) as a white solid.

[0339] [M+H + ] + =782.5. [Example]

[0340] Activity after membrane permeabilization Assay testing was performed as detailed in Example 20, and IRF pathway activation was measured between compound XV of Table 1 versus compound XX of Table 1, as detailed in Example 21, demonstrating activity against membrane permeabilization. [ka] Compound XV Compound XX

[0341] The main difference is the reduced hydrophobicity of compound XX as a result of the introduction of an oxazolyl heterocyclic ring in place of the pyrazolyl heterocyclic ring of compound XV and an imidazopyridine ring in place of the benzimidazole ring of compound XV. This substitution results in a 1-log decrease in cLogP and slightly better solubility, as shown in Table 5.

[0342] [Table 10] JPEG2026508759000189.jpg43170JPEG2026508759000190.jpg13170

[0343] EC50 values ​​were calculated using GraphPad Prism 9. Plots of IRF pathway activation for compounds XV and XX compared to diABZI are shown in Figure 10. [Example]

[0344] STING allele activity The assay was performed as described in Example 21. The biological activity of compounds XV and XX was evaluated as described in Example 20. The STING haplotype curves of compounds XV and XX are shown in Figure 11. [Example]

[0345] PBMC assay Compounds XV and XX were further tested on PBMCs from healthy donors to determine dose-dependent cytokine induction as described in Example 23. Representative curves of PBMCs for compounds XV and XX are shown in Figure 12.

[0346] Materials and Methods (Examples 33-44) LCMS: System I Waters HPLC-System CTC Pal Autosampler. One Waters 1525 Multisolvent Delivery System with a 10 μl sample loop. Waters Micromass ZQ single quadrupole mass spectrometer equipped with an electrospray source. MS method: positive / negative ion mode scan, m / z 80–800 or 80–2000, 1 s; capillary voltage, 3.0 kV; cone voltage, 20–50 V; multiplier voltage, 700 V; probe and desolvation gas temperatures, 120°C and 300°C, respectively. Waters 996 PDA detector set at 254 nm. Software, Waters Masslynx V 4.1.

[0347] LCMS System II Shimadzu LCMS-2050: SCL-40 system controller; DGU-405 degassing unit; 2 × LC-40D XR solvent delivery pumps; SUL-40C XR autosampler; CTO-40C column oven. MS LCMS-2050: Single quadrupole mass spectrometer with ESI / APCI DUIS source. MS method: Positive / negative ion mode scanning, m / z 100–2000, 0.45 s per event; desolvation temperature: 450°C; desolvation line temperature: 200°C; interface voltages: +3.0 kV and -2.0 kV. PDA detector: Shimadzu SPD-M40; start wavelength: 190 nm; end wavelengths: 600 nm, 254 nm, 200 nm, 220 nm, and 260 nm; cell temperature: 40°C. Software: Shimadzu LabSolutions Version 5.118.

[0348] Method A: 4.3 minutes

[0349] Column: Waters Phenomenex Onyx Monolythic C18 50 x 2 mm with stainless steel 2 μm prefilter

[0350] UV detector: 254 nm

[0351] Column temperature: room temperature

[0352] Pump Style: Gradient

[0353] [Table 11] JPEG2026508759000192.jpg49170

[0354] Method B: 7.5 minutes

[0355] Column: Waters Phenomenex Onyx Monolythic C18 50 x 2 mm with stainless steel 2 μm prefilter

[0356] UV detector: 254 nm

[0357] Column temperature: room temperature

[0358] Pump Style: Gradient

[0359] [Table 12] JPEG2026508759000194.jpg44170

[0360] Method C: 10 minutes

[0361] Column: Waters Phenomenex Onyx Monolythic C18 50 x 2 mm with stainless steel 2 μm prefilter

[0362] UV detector: 254 nm

[0363] Column temperature: room temperature

[0364] Pump Style: Gradient

[0365] [Table 13] JPEG2026508759000196.jpg44170

[0366] Method D: 15 minutes

[0367] Column: Waters Phenomenex Onyx Monolythic C18 50 x 2 mm with stainless steel 2 μm prefilter

[0368] UV detector: 254 nm

[0369] Column temperature: room temperature

[0370] Pump Style: Gradient

[0371] [Table 14] JPEG2026508759000198.jpg44170

[0372] HPLC The Dionex Ultimate 3000 was equipped with a solvent rack and a vacuum degasser SRD-3600, a binary pump DPG-3600SD, and an autosampler WPS-3000TFC: 15.0 μL needle volume, 50 μL sample loop, 6.2 μL bridge tube, and 250 μL syringe. The column compartment was a TCC-3200; the detector was a DAD-3000; the CAD-detector was Corona Veo. The HPLC software was Chromeleon Datasystem 7.3.

[0373] Method A

[0374] Column: Waters Phenomenex Onyx Monolythic C18 2 mm x 50 mm

[0375] UV detector: 190-400nm

[0376] Injection volume: 10μL

[0377] Column temperature: 30℃

[0378] Pump Style: Gradient

[0379] [Table 15] JPEG2026508759000200.jpg40170

[0380] Method B

[0381] Column: Waters XSelect CSH C18, 130Å, 3.5μm, 4.6mm x 100mm

[0382] UV detector: 190-400nm

[0383] Injection volume: 10μL

[0384] Column temperature: 30℃

[0385] Pump Style: Gradient

[0386] [Table 16] JPEG2026508759000202.jpg36170

[0387] Method C

[0388] Column: Waters XBridge BEH C18, 130Å, 3.5μm, 2.1mm x 100mm

[0389] UV detector: 190-400nm

[0390] Injection volume: 10μL

[0391] Column temperature: 30℃

[0392] Pump Style: Gradient

[0393] [Table 17] JPEG2026508759000204.jpg45170

[0394] Preparative HPLC Waters Autopurification System: Waters 3767 Autosampler (with 5 mL syringe and 10 mL sample loop), Waters System Fluid Organizer, Waters 2525 Binary Gradient Module, Waters 515 Make-Up Pump (50% acetonitrile in water + 0.1% formic acid, 1 mL / min), Waters 515 At-Column-Dilution Pump (different solvent mixtures, individually adapted to the sample, 5 mL / min for the first 2 minutes of the gradient), Waters 2998 Photo Diode Array Detector, Waters QDA Mass Spectrometer (for mass-triggered fractionation, scan mode, positive and negative polarization). Software: Waters MassLynx V 4.2.

[0395] Method I

[0396] Column: Waters Atlantis T3 OBD Prep Column, 100Å, 5μm, 19mm x 150mm, product number: 186003698, flow rate: 20mL / min

[0397] [Table 18] JPEG2026508759000206.jpg41170

[0398] Method II

[0399] Column: Waters Atlantis T3 OBD Prep Column, 100Å, 5μm, 19mm x 150mm, product number: 186003698, flow rate: 20mL / min

[0400] [Table 19] JPEG2026508759000208.jpg41170

[0401] Method III

[0402] Column: Waters XBridge BEH C18 OBD Prep Column, 130Å, 5μm, 19mm x 150mm, product number: 186002979, flow rate 20mL / min

[0403] [Table 20] JPEG2026508759000210.jpg41170

[0404] NMR NMR spectra were collected at 302 K (approximately 29 °C) on a Bruker UltraShield 300 MHz spectrometer equipped with a Nanobay AV III console, a B-ACS 60 autosampler, and a PH BBI 300 S1 H-BB-D-05 Z-probe. Deuterated solvents were used as described in the Experimental Procedures section. All spectra were processed and analyzed using TopSpin 3.6.0 and / or MestreNova 14.2.

[0405] Synthesis of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate [ka]

[0406] 1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (3.50 g, 22.7 mmol) was suspended in anhydrous CHCl (60 mL) under argon. Oxalyl chloride (2.4 mL, 27.2 mmol) was then added dropwise at room temperature, followed by DMF (170 μL, 2.27 mmol) over 1 minute at room temperature. The mixture was stirred at room temperature under argon for 18 hours. The reaction mixture was then evaporated to give 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride as a yellow oil. Potassium thiocyanate (2.87 g, 29.5 mmol) was suspended in acetone (25 mL) under argon. The suspension was then cooled to 0 °C in an ice bath. The previously obtained 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl chloride was dissolved in dry acetone (45 mL) under argon and then slowly added to the reaction mixture via a dropping funnel over 45 minutes. The resulting suspension was stirred at 0°C for 1.5 hours. Check by TLC (n-hexane / EtOAc = 9:1) showed the product. The solvent was removed under reduced pressure. The residue was suspended in n-hexane (50 mL) and sonicated. The solid was then filtered off and washed thoroughly with n-hexane. The filtrate was evaporated to give the crude product as a yellow liquid. Purification by automated flash column chromatography on silica: Biotage Selekt (Biotage Sfar Silica HC, 20 μM, 25 g, 80 mL / min, gradient from 3 CV n-hexane to 10 CV n-hexane to n-hexane / EtOAc 9:1 to 10 CV n-hexane / EtOAc 9:1) gave 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (3.67 g, 18.8 mmol, 83%) as a pale yellow liquid, which was stored in the freezer until use. 1 H NMR (CDCl3,300MHz): 1.32(t,3H,J=7.2Hz), 2.21(d,3H,J=0.6Hz), 4.42(q,2H,J=7.2Hz), 6.65(q,1H,J=0.7Hz).

[0407] Synthesis of 4-ethyl-2-methyloxazole-5-carbonyl isothiocyanate [ka]

[0408] 4-Ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (5.00 g, 32.2 mmol) was suspended in anhydrous CHCl (80 mL) under argon. Oxalyl chloride (3.44 mL, 38.7 mmol) was then added dropwise at room temperature, followed by DMF (248 μL, 3.22 mmol) over 1 minute at room temperature. The mixture was stirred under argon at room temperature for 3 hours. The solvent was removed under reduced pressure to give crude 4-ethyl-2-methyloxazole-5-carbonyl chloride. Potassium thiocyanate (4.07 g, 41.9 mmol) was suspended in acetone (25 mL) under argon. The suspension was cooled to 0° C., and a suspension of the crude 4-ethyl-2-methyloxazole-5-carbonyl chloride (5.59 g, 32.2 mmol) obtained above in acetone (50 mL) was added dropwise via an addition funnel over 10 minutes. The resulting mixture was stirred at 0 °C for 3 h. Product formation was monitored by TLC (EtOAc / n-hexane: 1 / 9). The solvent was removed under vacuum (cold trap). The residue was purified by automated flash column chromatography on silica using Biotage Selekt (Biotage Sfar HC 50 g, loading: solid (Extrelut), 120 mL / min, 0 to 20% EtOAc in n-hexane in 30 min) to give 4-ethyl-2-methyloxazole-5-carbonyl isothiocyanate (3.61 g, 18.3 mmol, 57%) as a yellow liquid, which was stored in the freezer until use. 1 H NMR (CDCl3,300MHz): 1.27(t,3H,J=7.6Hz), 2.54(s,3H), 2.90(q,2H,J=7.5Hz). [Example]

[0409] Synthesis scheme of compound XXVIII [ka]

[0410] Preparation of tert-butyl ((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)carbamate (XX6.1) [ka]

[0411] Compound XX5.4 (2.0 g, 5.1 mmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (949 mg, 6.11 mmol) were dissolved in DMF (15 mL). HATU (2.9 g, 7.6 mmol) and DIPEA (4.48 mL, 25.4 mmol) were then added, and the reaction mixture was stirred at room temperature for 30 minutes. Product formation was monitored by LC / MS. The mixture was quenched with water (30 mL), and the resulting solid was filtered off and washed with water. The product was dried in vacuo to give 1.75 g (3.30 mmol, 64%) of an off-white solid. LCMS (Method A) retention time 2.58 minutes, [M+H]+=530.1. 1H-NMR (300MHz, DMSO-d6) δ[ppm]=8.05 (brs,1H), 8.01~7.96(m,1H), 7.65~7.54(m,1H), 7.47(brs,1H), 6.89(t,J=5. 9Hz,1H), 5.82~5.55(m,2H), 5.29(d,J=5.5Hz,2H), 3.98(s,3H), 3.50(t,J=5. 5Hz,2H), 3.05~2.86(m,2H), 2.46(s,3H), 1.30(s,9H), 1.19(t,J=6.5Hz,3H).

[0412] Preparation of N-((Z)-3-((E)-4-aminobut-2-en-1-yl)-6-carbamoyl-4-methoxybenzo[d]thiazol-2(3H)-ylidene)-4-ethyl-2-methyloxazole-5-carboxamide hydrochloride (XX6.2) [ka]

[0413] Compound XX6.1 (1.75 g, 3.30 mmol) was suspended in 4 M HCl in dioxane (26.4 mL, 105 mmol). The resulting suspension was stirred at room temperature for 30 minutes. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 1.4 g (3.3 mmol, 100%) of an off-white solid, which was used without further purification. LCMS (method A) retention time 1.28 min, [M+H]+=430.1. 1H-NMR (300MHz, DMSO-d6) δ[ppm]=8.13(brs,1H), 8.06~7.93(m,3H), 7.68~7.58(m,1H), 7.58~7.43(m,1H), 6. 15~5.99(m,1H), 5.82~5.57(m,1H), 5.37(d,J=5.6Hz,2H), 4.02(s,3H), 3. 52~3.35(m,2H), 2.99(q,J=7.5Hz,2H), 2.48(s,3H), 1.22(t,J=7.5Hz,3H).

[0414] Preparation of tert-butyl 4-(5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)butanoate (XX6.3) [ka]

[0415] Compound XX6.2 (1.40 g, 3.00 mmol) was suspended in n-butanol (36 mL), DIPEA (2.58 mL, 15.1 mmol), and sodium bicarbonate (505 mg, 6.01 mmol), and the mixture was stirred at room temperature for 10 minutes. Then, tert-butyl 4-(5-carbamoyl-2-chloro-3-nitrophenoxy)butanoate (1.08 g, 3.00 mmol) was added to the mixture, and the reaction was stirred at 130 °C for 26 hours and at room temperature for 18 hours. Product formation was monitored by LC / MS. The reaction was cooled to room temperature and diluted with water (50 mL). The aqueous layer was extracted with CHCl / CHOH (9:1, 70 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure and the residue was purified by automated flash column chromatography on silica using a Biotage Selekt (Biotage Sfar HC 50 g, 120 mL / min, 0-20% CHOH in CHCl over 30 min). Fractions containing the product were combined to give 603 mg (802 μmol, 27%) of a yellowish solid after solvent removal. LCMS (method B) retention time 3.46 minutes, [M+H]+=752.2. 1H-NMR (300MHz, DMSO-d6) δ[ppm]=8.08~8.01(m,2H), 7.98~7.94( m,1H), 7.91(brs,1H), 7.63(t,J=6.3Hz,1H), 7.57~7.53(m,1H), 7.46(brs,1H), 7.42~7.39(m,1H), 7.26( brs,1H), 5.85~5.61(m,2H), 5.26(d,J=5.2Hz,2H), 4.10(t,J=5.8Hz,2H), 3.94~3.82(m,5H), 2.89(q,J=7 .5Hz,2H), 2.44(s,3H), 2.26(t,J=7.4Hz,2H), 1.84(p,J=6.8Hz,2H), 1.35(s,9H), 1.13(t,J=7.5Hz,3H).

[0416] Preparation of tert-butyl 4-(3-amino-5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)amino)phenoxy)butanoate (XX6.4) [ka]

[0417] Compound XX6.3 (600 mg, 798 μmol) was dissolved in CHOH (15 mL) and cooled to 0 °C. Sodium dithionite (1.39 g, 7.98 mmol) dissolved in water (2 mL) was then added. NH (30% aqueous solution, 790 μL, 5.99 mmol) was then added to this mixture at 0 °C. The reaction was allowed to warm to room temperature and stirred at room temperature for 3 h. NH (30% aqueous solution, 211 μL, 1.60 mmol) was added again, and the reaction was stirred at room temperature for 16 h. NH (30% aqueous solution, 211 μL, 1.60 mmol) and sodium dithionite (280 mg, 1.60 mmol) were then added, and the reaction was stirred at room temperature for 3 h. The progress of the reaction was monitored by LC / MS. The reaction mixture was diluted with HO (75 mL), and then CHCl (100 mL) was added. The aqueous layer was extracted with CH2Cl2 / CH3OH (8:2, 50 mL). The combined organic layers were dried over sodium sulfate and filtered. The solvent was removed under reduced pressure to give 450 mg (623 μmol, 78%) of a white solid, which was used without further purification. LCMS (Method A) retention time 1.94 min, [M+H] + =722.4. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=8.05(brs,1H), 8.00~7.93(m,1H), 7.60~7.51(m,2H), 7.44(b rs,1H), 6.92(brs,1H), 6.86~6.77(m,1H), 6.73~6.65(m,1H), 5.89~5.60(m,2H), 5.28(d,J=5.4 Hz,2H), 4.59(s,2H), 3.92~3.85(m,4H), 3.79(t,J=6.3Hz,2H), 3.56(s,2H), 2.95(q,J=7.6Hz, 2H), 2.45(s,3H), 2.25(t,J=7.3Hz,2H), 1.87~1.71(m,2H), 1.34(s,9H), 1.15(t,J=7.5Hz,3H).

[0418] Preparation of tert-butyl 4-((2-amino-5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX6.5) [ka]

[0419] Compound XX6.4 (450 mg, 623 μmol) was dissolved in CHOH (10 mL) and cooled to 0° C. Then, BrCN (132 mg, 1.24 mmol) was added. The reaction mixture was stirred at room temperature for 5 h. Again, BrCN (33 mg, 0.31 mmol) was added, and the mixture was stirred at room temperature for 19 h. Then, BrCN (33 mg, 0.31 mmol) was added, and the reaction was stirred at room temperature for 2.5 h. Product formation was monitored by LC / MS. The solvent was removed under reduced pressure. The residue was suspended in petroleum ether, and the resulting solid was filtered off. The product was dried in vacuo to give 436 mg (583 μmol, 93%) of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 2.05 min, [M+H] + =747.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=8.57(brs,2H), 8.11~8.01(m,2H), 8.01~7.97(m,1H), 7 .61~7.53(m,1H), 7.52~7.38(m,3H), 7.37~7.31(m,1H), 5.94~5.66(m,2H), 5.30(d,J=5. 3Hz,2H), 4.86(d,J=5.1Hz,2H), 3.99(t,J=6.5Hz,2H), 3.81(s,3H), 2.79(q,J=7.5Hz,2H) ), 2.43(s,3H), 2.29~2.11(m,2H), 1.92~1.70(m,2H), 1.35(s,9H), 1.04(t,J=7.5Hz,3H).

[0420] Preparation of tert-butyl 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-2-(4-ethyl-2-methyloxazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX6.6) [ka]

[0421] Compound XX6.5 (425 mg, 569 μmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (132 mg, 853 μmol) were dissolved in DMF (5 mL). HATU (327 mg, 853 μmol) and DIPEA (150 μL, 853 μmol) were then added, and the reaction mixture was stirred at room temperature for 4 hours. 4-Ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (44.1 mg, 284 μmol) was added again, and the reaction mixture was stirred at room temperature for 2 hours. DIPEA (50.0 μL, 284 μmol) was then added, and the reaction mixture was stirred at room temperature for 21 hours. DIPEA (100 μL, 568 μmol) was added again, and the reaction mixture was stirred at room temperature for 2 hours. HATU (109 mg, 284 μmol) was then added, and the mixture was stirred at room temperature for 19 hours. Product formation was monitored by LC / MS. The solvent was removed under reduced pressure. The mixture was quenched with water (15 mL), and the solid was filtered off and washed with water. The product was dried in vacuo. The residue was suspended again in water, filtered off, and dried in vacuo to give 377 mg (426 μmol, 75%) of an orange solid, which was used without further purification. LCMS (Method A) retention time 2.31 min, [M+H] + =884.3.

[0422] Preparation of 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-2-(4-ethyl-2-methyloxazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoic acid (XX6.7) [ka]

[0423] Compound XX6.6 (365 mg, 413 μmol) was suspended in dioxane (2 mL) and 4 M HCl in dioxane (3.3 mL, 13.2 mmol) was added. The resulting suspension was stirred at room temperature for 1 hour. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 310 mg (358 μmol, 87%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 1.94 min, [M+H] + =828.3.

[0424] Preparation of N-(5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-7-(4-(1,2-dimethylhydrazinyl)-4-oxobutoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methyloxazole-5-carboxamide (Compound XXVIII) [ka]

[0425] Compound XX6.7 (300 mg, 347 μmol) was suspended in DMF (5 mL). Then, DIPEA (181 μL, 1.04 mmol), NHS (81.5 mg, 694 μmol), and DIC (107 μL, 694 μmol) were added, and the resulting reaction mixture was stirred at 40° C. for 4 hours. NHS (81.5 mg, 694 μmol) was added again, and the mixture was stirred at 40° C. for 17.5 hours. The progress of the reaction was monitored by LC / MS. Then, 1,2-dimethylhydrazine dihydrochloride (231 mg, 1.73 mmol) and DIPEA (181 μL, 1.04 mmol) were added, and the resulting reaction mixture was stirred at 40° C. for 4.5 hours. DIPEA (60.4 μL, 347 μmol) was added again, and the reaction mixture was stirred at 40° C. for 19 hours. DIPEA (302 μL, 1.74 mmol) was then added and the reaction was stirred at 40° C. for 6 h. The reaction progress was monitored by LC / MS. The solvent was removed under reduced pressure and the residue was purified by preparative HPLC (Method II) on RP18 silica. The fractions containing the product were freeze-dried to give 4 mg (4.1 μmol, 90.9% pure by HPLC, Method C) and 21 mg (21 μmol, 84.5% pure by HPLC Method C), respectively, as white solids. LCMS (Method B) retention time 2.74 min, [M+H] + =870.2. [Example]

[0426] Synthesis scheme of compound XXIX [ka]

[0427] Preparation of methyl (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (XX8.1) [ka]

[0428] Compound XX7.4 (2.7 g, 7.1 mmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (1.3 g, 8.5 mmol) were dissolved in DMF (20 mL). HATU (4.1 g, 11 mmol) and DIPEA (6.2 mL, 35 mmol) were then added, and the reaction mixture was stirred at room temperature for 30 minutes. The mixture was quenched with water (40 mL), and the solid was filtered off and washed with water. The product was dried in vacuo to give 3.5 g (6.7 mmol, 97%) as an off-white solid, which was used without further purification. LCMS (Method A) retention time 3.57 min, [M+H] + =516.1. 1 H-NMR(300MHz,CDCl3)δ[ppm]=9.11~9.01(m,1H), 8.59~8.51(m,1H), 5.86(t,J=3.5Hz,2H), 5.19~5.11(m,2H), 4.50( brs,1H), 3.98(s,3H), 3.73(d,J=5.6Hz,2H), 3.10(q,J=7.5Hz,2H), 2.55(s,3H), 1.39(s,9H), 1.32(t,J=7.5Hz,3H).

[0429] Preparation of (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylic acid (XX8.2) [ka]

[0430] Compound XX8.1 (3.5 g, 6.7 mmol) was dissolved in CHOH / THF / HO (2:2:1, 50 mL). LiOH (487 mg, 20.3 mmol) was then added, and the resulting reaction mixture was stirred at room temperature for 18 h. The organic solvent was removed in vacuo (rotary evaporator). The aqueous phase was cooled to 0 °C and neutralized with 1 M aqueous HCl. The precipitate was filtered and washed with water. The solid was dissolved / suspended in CHCN / HO (1:1, 25 mL) and freeze-dried to give 2.16 g (4.31 μmol, 63%) of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 2.27 min, [M+H] + =502.0. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=9.01~8.93(m,1H), 8.88~8.78(m,1H), 6.89(brs,1H), 5.84~5.55(m,2H), 5.05 (d,J=5.2Hz,2H), 3.49(d,J=5.6Hz,2H), 3.01(q,J=7.5Hz,2H), 2.48(s,3H), 1.27(s,9H), 1.22(t,J=7.5Hz,3H).

[0431] Preparation of tert-butyl ((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)carbamate (XX8.3) [ka]

[0432] Compound XX8.2 (2.0 g, 4.0 mmol) was dissolved in DMF (8 mL), followed by the addition of NH4Cl (639 mg, 12.0 mmol), HATU (2.2 g, 6.0 mmol), and DIPEA (2.0 mL, 12 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The mixture was diluted with water (30 mL), and the resulting suspension was filtered. The solid was washed with water and dried in vacuo to give 1.59 g (3.18 mmol, 79%) of a white solid, which was used without further purification. LCMS (Method A) retention time 2.11 min, [M+H] + =501.0. 1 H-NMR(300MHz,DMSO-d6)d:9.02~8.92(m,1H), 8.78~8.72(m,1H), 8.17(brs,1H), 7.62(brs,1H), 6.89(t,J=6.0Hz,1H), 5.84~5. 60(m,2H), 5.05(d,J=5.2Hz,2H), 3.50(t,J=5.5Hz,2H), 3.01(q,J=7.5Hz,2H), 2.48(s,3H), 1.30(s,9H), 1.22(t,J=7.5Hz,3H).

[0433] Preparation of N-((Z)-3-((E)-4-aminobut-2-en-1-yl)-6-carbamoylthiazolo[4,5-b]pyridin-2(3H)-ylidene)-4-ethyl-2-methyloxazole-5-carboxamide (XX8.4) [ka]

[0434] Compound XX8.3 (1.7 g, 3.4 mmol) was suspended in dioxane and 4 M HCl (25.4 mL, 102 mmol) was added. The resulting suspension was stirred at room temperature for 1 h. The formed solid was filtered off, washed with EtO, and dried in vacuo. The residue was purified by automated flash column chromatography on a Buchi C-850 (XBridge® Prep OBD™ C18 0.5 μM, 50 mm × 250 mm, 100 mL / min; 5 to 100% CH3CN in HO (50 μM NH4HCO3) in 30 min). The product-containing fractions were combined to give 1.10 g (2.75 mmol, 81%) of a yellowish solid. LCMS (Method A) retention time 1.30 min, [M+H] + =401.0. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.95(d,J=2.0Hz,1H), 8.74(d,J=1.9Hz,1H), 8.17(brs,1H), 7.62(brs,1H), 5. 83~5.75(m,2H), 5.09~5.02(m,2H), 3.15~3.08(m,2H), 3.02(q,J=7.5Hz,2H), 2.48(s,3H), 1.22(t,J=7.5Hz,3H).

[0435] Preparation of tert-butyl 4-(5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)butanoate (XX8.5) [ka]

[0436] Compound XX8.4 (220 mg, 549 μmol) was suspended in n-butanol (6 mL). Then, DIPEA (483 μL, 2.74 mmol) and sodium bicarbonate (92 mg, 1.1 mmol) were added, and the mixture was stirred at room temperature for 10 minutes. tert-Butyl 4-(5-carbamoyl-2-chloro-3-nitrophenoxy)butanoate (197 mg, 549 μmol) was then added, and the reaction mixture was stirred at 130 °C for 18 hours. The reaction was cooled to room temperature and quenched with water (30 mL). The aqueous layer was extracted with CHCl / CHOH (9:1, 70 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure and the resulting residue was purified by automated flash column chromatography on silica using a Biotage Selekt (Biotage Sfar HC 10 g, 40 mL / min, 0-20% CHOH in CH2Cl2 over 30 min). Product-containing fractions were combined and the solvent evaporated to give 230 mg (318 μmol, 57%) of an orange solid. LCMS (Method B) retention time 4.53 min, [M+H] + =723.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.91(d,J=2.0Hz,1H), 8.73(d,J=1.9Hz,1H), 8.15(s,1H), 8. 02(d,J=1.9Hz,1H), 7.89(s,1H), 7.67~7.57(m,2H), 7.40(d,J=1.9Hz,1H), 7.26(s,1H), 5.81~5 .73(m,2H), 5.04~4.98(m,2H), 4.12~4.01(m,2H), 3.90(t,J=6.3Hz,2H), 2.90(q,J=7.5Hz,2H) , 2.46(s,3H), 2.28(t,J=7.3Hz,2H), 1.85(p,J=6.8Hz,2H), 1.36(s,9H), 1.13(t,J=7.5Hz,3H).

[0437] Preparation of tert-butyl 4-(3-amino-5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)amino)phenoxy)butanoate (XX8.6) [ka]

[0438] Compound XX8.5 (735 mg, 1.02 mmol) was dissolved in MeOH (20 mL) and cooled to 0 °C. Sodium dithionite (1.7 g, 10.2 mmol) in water (5 mL) and aqueous NH (30% aqueous solution, 1.0 mL, 7.6 mmol) were then added. The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was diluted with HO (75 mL), and then CHCl (100 mL) was added. The aqueous layer was extracted with CHCl / MeOH (8:2, 2 × 50 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure to give 591 mg (853 μmol, 84%) of an orange solid, which was used without further purification. LCMS (Method A) retention time 1.93 min, [M+H] + =693.3. 1 H-NMR (300MHz, DMSO-d6): δ[ppm]=8.93(d,J=2.0Hz,1H), 8.74(d,J=2.0Hz,1H), 8.17(brs,1H), 7.71~7.46(m,2H), 6.92(brs,1H), 6.80(d,J=1.8Hz,1H), 6.68(d,J=1.8Hz,1H), 5.93~5.70(m,2 H), 5.03(d,2H), 4.60(s,2H), 3.83(t,J=6.3Hz,2H), 3.54(d,J=4.8Hz,2H), 2.98(q,J=7.5Hz,2H ), 2.47(s,3H), 2.28(t,J=7.3Hz,2H), 1.81(p,J=6.8Hz,2H), 1.34(s,9H), 1.18(t,J=7.5Hz,3H).

[0439] Preparation of tert-butyl 4-((2-amino-5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX8.7) [ka]

[0440] Compound XX8.6 (575 mg, 830 μmol) was dissolved in MeOH (15 mL) and cooled to 0° C. Then, BrCN (175 mg, 1.66 mmol) was added, and the reaction mixture was stirred at room temperature for 5 hours. The solvent was removed under reduced pressure. The residue was suspended in petroleum ether and filtered off. The product was dried in vacuo to give 540 mg (752 μmol, 91%) of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 1.72 min, [M+H] + =718.2. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.93(d,J=1.9Hz,1H), 8.77(d,J=1.9Hz,1H), 8.38(brs,2H), 8.17 (brs,1H), 8.00(brs,1H), 7.63(brs,1H), 7.45(d,J=1.2Hz,1H), 7.37(brs,1H), 7.30(d,J=1.3Hz,1H ), 5.93~5.77(m,2H), 5.09~5.01(m,2H), 4.86~4.80(m,2H), 3.99(t,J=6.3Hz,2H), 2.76(q,J=7.5Hz ,2H), 2.44(s,3H), 2.22(t,J=7.3Hz,2H), 1.77(p,J=6.9Hz,2H), 1.34(s,9H), 1.01(t,J=7.5Hz,3H).

[0441] Preparation of tert-butyl 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-2-(4-ethyl-2-methyloxazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX8.8) [ka]

[0442] Compound XX8.7 (520 mg, 724 μmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (112 mg, 724 μmol) were dissolved in DMF (6 mL). HATU (417 mg, 1.09 mmol) and DIPEA (191 μL, 1.09 mmol) were then added, and the reaction mixture was stirred at room temperature for 22 hours. 4-Ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (56.2 mg, 362 μmol), HATU (278 mg, 724 μmol), and DIPEA (127 μL, 724 μmol) were added again, and the reaction mixture was stirred at room temperature for 20 hours. Product formation was monitored by LC / MS. The mixture was quenched with water (10 mL), and the solid was filtered off and washed with water. The product was dried in vacuo, and the residue was suspended in acetonitrile, filtered, and dried. The solid was purified by preparative TLC (2 mm plate, CHCl / CHOH = 9:1). The filtrate containing the product was purified by flash column chromatography using a Buchi C-850 (XSelect® CSH™ Prep OBD™ C18 0.5 μM, 50 mm × 150 mm, 100 mL / min, 5 to 100% CHCN in HO (+0.15% TFA) in 30 min) to give 115 mg (134 μmol, 18%) of a yellowish solid. LCMS (Method A) retention time 2.20 min, [M+H] + =855.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=12.69(brs,1H), 8.91(d,J=2.0Hz,1H), 8.74(d,J=1.9Hz,1H), 8 .14(brs,1H), 7.92(brs,1H), 7.65~7.59(m,2H), 7.31~7.25(m,2H), 5.98~5.83(m,1H), 5.82~5.67 (m,1H), 5.07~4.99(m,2H), 4.90(d,J=5.0Hz,2H), 4.02(t,J=6.3Hz,2H), 2.85~2.68(m,4H), 2.42( s,3H), 2.39(s,3H), 2.26(t,J=7.3Hz,2H), 1.82(p,J=6.9Hz,2H), 1.35(s,9H), 1.05~0.91(m,6H).

[0443] Preparation of 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-2-(4-ethyl-2-methyloxazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoic acid (XX8.9) [ka]

[0444] Compound XX8.8 (60.0 mg, 70.2 μmol) was suspended in dioxane (2 mL) and 4 M HCl in dioxane (560 μL, 2.25 mmol) was added. The resulting suspension was stirred at room temperature for 20 minutes. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 42 mg (50 μmol, 72%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 1.89 min, [M+H] + =799.5. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=12.70(brs,1H), 8.92(d,J=2.0Hz,1H), 8.81~8.71(m, 1H), 8.15(s,1H), 7.93(s,1H), 7.66~7.59(m,2H), 7.33~7.27(m,2H), 5.99~5.70(m,2H) , 5.04(d,J=5.4Hz,2H), 4.91(d,J=5.1Hz,2H), 4.05(t,J=6.3Hz,2H), 2.85~2.69(m,4H) , 2.43(s,3H), 2.41(s,3H), 2.31(t,J=7.2Hz,2H), 1.92~1.77(m,2H), 1.05~0.92(m,6H).

[0445] Preparation of N-(5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-7-(4-(1,2-dimethylhydrazinyl)-4-oxobutoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methyloxazole-5-carboxamide (Compound XXIX) [ka]

[0446] Compound XX8.9 (50.0 mg, 59.9 μmol) was suspended in DMF (3 mL). Then, DIPEA (31.2 μL, 179 μmol), NHS (14.1 mg, 119 μmol), and DIC (18.5 μL, 119 μmol) were added, and the resulting reaction mixture was stirred at 45 °C for 24 h. The reaction progress was monitored by LC / MS. 1,2-Dimethylhydrazine dihydrochloride (39.8 mg, 299 μmol) and DIPEA (104 μL, 598 μmol) were then added, and the resulting reaction mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LC / MS. The solvent was removed under reduced pressure, and the residue was purified by preparative HPLC (Method II) on RP18 silica gel. The product-containing fractions were freeze-dried to give 19 mg (20 μmol, 33%, 95.9% purity by HPLC Method A) of a white solid. LCMS (Method D) retention time 4.24 min, [M+H] + =842.2. [Example]

[0447] Synthesis scheme of compound XXX [ka]

[0448] Preparation of 3-methoxy-4-nitrobenzamide (XX5.1) [ka]

[0449] To a stirred solution of 3-methoxy-4-nitrobenzoic acid (11.3 g, 57.3 mmol) in DMF (100 mL) was added NH4Cl (9.2 g, 172.0 mmol), HATU (32.7 g, 86.0 mmol), and DIPEA (29.4 mL, 172.0 mmol). The mixture was stirred at room temperature for 1 hour. The mixture was diluted with water. A precipitate formed, which was filtered off and washed with water. The solid was dried in vacuo to give 10.4 g (53.0 mmol, 93%) of a white solid. LCMS (Method A) retention time 1.85 min, [M+H] + =197.0.1 H-NMR (300MHz, DMSO-d6) δ[ppm]=8.22(s,1H), 7.94(d,J=8.3Hz,1H), 7.75(d,J=1.6Hz,1H), 7.68(s,1H), 7.57(dd,J=8.3,1.7Hz,1H), 3.98(s,3H).

[0450] Preparation of 4-amino-3-methoxybenzamide (XX5.2) [ka]

[0451] 3-Methoxy-4-nitrobenzamide (10.4 g, 53.0 mmol) was dissolved in CHOH (200 mL) and 10% Pd—C (1.9 g, 1.8 mmol) was added under an inert atmosphere. The resulting mixture was stirred under an H atmosphere at room temperature for 18 hours. The catalyst was removed by filtration through a small pad of CELITE® Hyflo Supercel. The filter cake was washed with CHOH. The solvent was removed under reduced pressure. The product was dried in vacuo to give 8.5 g (51.1 mmol, 96%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 1.06 min, [M+H] + =167.0. 1 H-NMR (300MHz, DMSO-d6) δ[ppm]: 7.57 (brs, 1H), 7.37~7.25 (m, 2H), 6.87 (brs, 1H), 6.59 (d, J = 8.0Hz, 1H), 5.21 (brs, 2H), 3.79 (s, 3H).

[0452] Preparation of tert-butyl (E)-(4-((4-carbamoyl-2-methoxyphenyl)amino)but-2-en-1-yl)carbamate (XX5.3) [ka]

[0453] 4-Amino-3-methoxybenzamide (2.5 g, 15 mmol) was dissolved in DMF (15 mL). CsCO (7.4 g, 22 mmol) and tert-butyl N-[(2E)-4-bromobut-2-en-1-yl]carbamate (3.7 g, 15 mmol) were then added, and the mixture was stirred at 60 °C for 1 h. The mixture was diluted with water (30 mL), and the product was extracted with ethyl acetate (50 mL). The organic solvent was removed under reduced pressure. The residue was suspended in diethyl ether and then filtered. The solid was washed with EtO and dried in vacuo to give 2.81 g (8.38 mmol, 56%) of a white solid, which was used without further purification. LCMS (Method A) retention time 2.33 min, [M+H] + =336.0. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]:7.59(brs,1H), 7.42~7.34(m,1H), 7.32(d,J=1.9Hz,1H), 6.91(brs,2H), 6.46(d,J= 8.3Hz,1H), 5.59~5.54(m,2H), 5.50(t,J=6.1Hz,1H), 3.82(s,3H), 3.78~3.71(m,2H), 3.58~3.47(m,2H), 1.36(s,9H).

[0454] Preparation of tert-butyl (E)-(4-(6-carbamoyl-2-imino-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)carbamate (XX5.4) [ka]

[0455] Compound XX5.3 (4.25 g, 12.7 mmol) was dissolved in acetic acid (14.0 mL) and KSCN (4.93 g, 50.7 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. Then, Br2 (649 μL, 12.7 mmol) was dissolved in acetic acid (4.5 mL) and added to the mixture. The reaction was stirred at room temperature for 30 minutes. The mixture was quenched with water (35 mL). The solid was filtered off. The filtrate was adjusted to pH 9 with aqueous ammonia (33%) and the aqueous layer was extracted with ethyl acetate (2 × 100 mL). The combined organic layers were dried over sodium sulfate and filtered. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography using a Biotage Select column (DCM / [DCM / MeOH (85:15)], 80:20 to 50:50, linear gradient, 120 mL / min, 30 min, Biotage Sfar Select HC 50 g). Product-containing fractions were combined and the solvent evaporated to give 2.02 g (5.15 mmol, 41%) of a beige solid. LCMS (Method A) retention time 1.35 min, [M+H] + =393.2. 1 H-NMR:(300MHz,DMSO-d6)δ[ppm]:8.39(brs,1H), 7.84(brs,1H), 7.53(d,J=1.5Hz), 7.41(d,J=1.6Hz), 7.25(brs, 1H), 6.91(t,J=5.9Hz,1H), 5.68~5.44(m,2H), 4.77(d,J=5.1Hz,2H), 3.86(s,3H), 3.53~3.44(m,2H), 1.34(s,9H).

[0456] Preparation of tert-butyl ((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)carbamate (XX5.5) [ka]

[0457] Compound XX5.4 (100 mg, 254 μmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (47 mg, 0.30 mmol) were dissolved in DMF (2 mL). HATU (147 mg, 382 μmol) and DIPEA (224 μL, 1.27 mmol) were then added, and the reaction mixture was stirred at room temperature for 2.5 hours. The mixture was quenched with water (15 mL), and the solid was filtered off and washed with water. The product was dried in vacuo to give 100 mg (189 μmol, 75%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 2.76 min, [M+H] + =529.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]:8.08(brs,1H), 8.00(d,J=1.4Hz,1H), 7.62(d,J=1.5Hz,1H), 7.47(brs,1H), 6.96~6.85(m,1H), 6.78 (s,1H), 5.84~5.61(m,2H), 5.39~5.31(m,2H), 4.58(q,J=7.1Hz,2H), 4.00(s,3H), 3.55~3.46(m,2H), 2.20(s,3H), 1.42~1.27(m,12H).

[0458] Preparation of (Z)-3-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxamide hydrochloride (XX5.6) [ka]

[0459] Compound XX5.5 (1.0 g, 1.9 mmol) was suspended and 4 M HCl in dioxane (15 mL, 61 mmol) was added. The resulting suspension was stirred at room temperature for 2 hours. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 860 mg (1.85 mmol, 98%) of an off-white solid. LCMS (Method A) retention time 1.38 min, [M+H] + =429.0. 1H-NMR(300MHz,DMSO-d6)δ[ppm]:8.16(s,4H), 8.06~8.00(m,1H), 7.70~7.64(m,1H), 6.16~6.00(m,1H), 5.85~5.69(m, 1H), 5.38(d,J=5.8Hz,2H), 4.57(q,J=7.1Hz,2H), 4.03(s,3H), 3.49~3.31(m,2H), 2.20(s,3H), 1.34(t,J=7.1Hz,3H).

[0460] Preparation of tert-butyl 4-(5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2i)-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)butanoate (XX5.7) [ka]

[0461] Compound XX5.6 (500 mg, 1.07 mmol) was suspended in n-butanol (15 mL), followed by the addition of DIPEA (945 μL, 5.37 mmol) and sodium bicarbonate (180 mg, 2.15 mmol). The resulting mixture was stirred at room temperature for 10 minutes. Then, tert-butyl 4-(5-carbamoyl-2-chloro-3-nitrophenoxy)butanoate (385 mg, 1.07 mmol) was added to the mixture, and the reaction was stirred at 130 °C for 18 hours. The reaction was cooled to room temperature and quenched with water (50 mL). The aqueous layer was extracted with CHCl / CHOH (9 / 1, 75 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure. The residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 25 g, 80 mL / min, 0-20% CHOH in CH2Cl2 over 30 min). Fractions containing the product were pooled and the solvent evaporated to give 299 mg (398 μmol, 37%) of a reddish solid. LCMS (Method A) retention time 3.72 min, [M+H] +=751.3. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]:8.08~8.02(m,2H), 8.00~7.95(m,1H), 7.93(brs,1H), 7.64( t,J=6.3Hz,1H), 7.59~7.54(m,1H), 7.46(s,1H), 7.43~7.37(m,1H), 7.26(s,1H), 6.72~6.66(m ,1H), 5.82~5.72(m,2H), 5.29(d,J=4.4Hz,2H), 4.55(q,J=7.1Hz,2H), 4.10(t,J=4.9Hz,2H), 3.92~3.82(m,5H), 2.25(t,J=7.4Hz,2H), 2.17(s,3H), 1.91~1.76(m,2H), 1.44~1.27(m,12H).

[0462] Preparation of tert-butyl 4-(3-amino-5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)amino)phenoxy)butanoate (XX5.8) [ka]

[0463] To a stirred solution of compound XX5.7 (900 mg, 1.20 mmol) in CHOH (20 mL) was added sodium dithionite (2.1 g, 12 mmol) dissolved in water (5 mL) at 0 °C. Then, NH (30% aqueous solution, 1.19 mL, 8.99 mmol) was added to the mixture at 0 °C. The reaction was warmed to room temperature and stirred at room temperature for 3.5 h. The reaction was diluted with HO (75 mL) and CHCl (100 mL) was added. The aqueous layer was extracted with CHCl / CHOH (8:2, 5 × 50 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure to give 695 mg of a yellowish solid (964 μmol, 80%). The residue was used without further purification. LCMS (Method A) retention time 2.07 min, [M+H] + =721.2. 1H-NMR (300MHz, DMSO-d6): δ[ppm]=8.08(s,1H), 7.98(d,J=1.4Hz,1H), 7.59(d,J=1.5Hz,1 H), 7.47(s,1H), 6.93(s,1H), 6.82(d,J=1.8Hz,1H), 6.74(d,J=0.6Hz,1H), 6.69(d,J=1.9H) z,1H), 5.91~5.66(m,2H), 5.32(d,J=5.2Hz,2H), 4.63~4.50(m,4H), 3.90(s,4H), 3.79(t,J =6.3Hz,2H), 3.62~3.52(m,3H), 2.30~2.14(m,5H), 1.86~1.71(m,2H), 1.37~1.31(m,12H).

[0464] Preparation of tert-butyl 4-((2-amino-5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX5.9) [ka]

[0465] Compound XX5.8 (660 mg, 915 μmol) was dissolved in CHOH (15 mL) and cooled to 0° C., after which BrCN (193 mg, 1.81 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. The formation of the product was controlled by LC / MS. The solvent was removed under reduced pressure. The residue was suspended in petroleum ether and the solid was filtered off. The product was dried in vacuo to give 737 mg of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 1.83 min, [M+H] + =746.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=8.55(brs,2H), 8.10~7.97(m,3H), 7.60~7.54(m,1H), 7.53~ 7.44(m,2H), 7.44~7.29(m,2H), 6.67~6.57(m,1H), 5.97~5.73(m,2H), 5.33(d,J=5.0Hz,2H), 4.86(d,J=4.9Hz,2H), 4.51(q,J=7.1Hz,2H), 3.97(t,J=6.4Hz,2H), 3.80(s,3H,CH3), 2.21(t ,J=7.3Hz,2H), 2.15~2.10(m,3H), 1.77(p,J=6.8Hz,2H), 1.34(s,9H), 1.29(t,J=7.1Hz,3H).

[0466] Preparation of tert-butyl 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX5.10) [ka]

[0467] Compound XX5.9 (430 mg, 576 μmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (133 mg, 865 μmol) were dissolved in DMF (5 mL). HATU (332 mg, 865 μmol) and DIPEA (152 μL, 865 μmol) were then added, and the reaction mixture was stirred at room temperature for 19 hours. HATU (110 mg, 288 μmol) and DIPEA (50 μL, 288 μmol) were added again, and the mixture was stirred at room temperature for 4 hours. DIPEA (100 μL, 576 μmol) was then added, and the reaction mixture was stirred at room temperature for 2 hours. 1-Ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (44 mg, 288 μmol) was added again, and the reaction mixture was stirred at room temperature for 18 hours. Product formation was monitored by LC / MS. The mixture was quenched with water (15 mL), and the solid was filtered off and washed with water. The product was dried in vacuo. The residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 10 g, 40 mL / min, 0-20% CHOH in CHCl over 30 min) followed by preparative TLC (CHCl / CHOH = 9:1). The resulting product was suspended in acetonitrile, and the solid was filtered off and dried in vacuo. The solid was suspended in methanol, and the solid was filtered off and dried in vacuo to give 89.0 mg (101 μmol, 17%) of an off-white solid. LCMS (Method A) retention time 2.50 min, [M+H] + =882.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=12.80(brs,1H), 8.04(brs,1H), 8.00~7.91(m,2H), 7.67~7.61(m,1H ), 7.56~7.50(m,1H), 7.46(brs,1H), 7.34~7.24(m,2H), 6.63~6.54(m,1H), 6.53~6.47(m,1H), 5.96~5 .81(m,2H), 5.31(d,J=4.7Hz,2H), 4.93(d,J=4.6Hz,2H), 4.50(q,J=7.0Hz,4H), 3.95(t,J=6.3Hz,2H) , 3.75(s,3H), 2.20(t,J=7.3Hz,2H), 2.10(s,6H), 1.81~1.71(m,2H), 1.34(s,9H), 1.30~1.15(m,6H).

[0468] Preparation of 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoic acid (XX5.11) [ka]

[0469] Compound XX5.10 (220 mg, 221 μmol) was suspended in dioxane (1 mL) and 4 M HCl in dioxane (1.77 mL, 7.07 mmol) was added. The resulting suspension was stirred at room temperature for 1 hour. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 118 mg (136 μmol, 62%) as a white solid. LCMS (Method A) retention time 2.05 min, [M+H] + =826.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=8.05(brs,1H), 7.99~7.93(m,1H), 7.67~7.61(m,1H), 7.56 ~7.50(m,1H), 7.45(s,1H), 7.33~7.27(m,2H), 6.59(s,1H), 6.48(s,1H), 5.98~5.74(m,2H), 5 .30(d,J=5.0Hz,2H), 4.93(d,J=4.8Hz,2H), 4.57~4.42(m,4H), 3.98(t,J=6.5Hz,2H), 3.76(s ,3H), 2.26(t,J=7.2Hz,2H), 2.09(d,J=1.6Hz,6H), 1.79(p,J=6.8Hz,2H), 1.37~1.19(m,6H).

[0470] Preparation of (Z)-3-((E)-4-(5-carbamoyl-7-(4-(1,2-dimethylhydrazinyl)-4-oxobutoxy)-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-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxamide (Compound XXX) [ka]

[0471] Compound XX5.11 (109 mg, 126 μmol) was suspended in DMF (2 mL). Then, DIPEA (66 μL, 0.37 mmol), NHS (30 mg, 0.25 mmol), and DIC (39 μL, 0.25 mmol) were added, and the resulting reaction mixture was stirred at room temperature for 2.5 h. NHS (30 mg, 0.25 mmol) was added again, and the mixture was stirred at 40 °C for 4 h and at room temperature for 13 h. The reaction progress was monitored by LC / MS. Then, 1,2-dimethylhydrazine dihydrochloride (84.1 mg, 632 μmol) and DIPEA (66.0 μL, 379 μmol) were added, and the resulting reaction mixture was stirred at room temperature for 18 h and at 40 °C for 2 h. The reaction progress was monitored by LC / MS. The solvent was removed under reduced pressure, and the residue was purified by preparative HPLC (Method I) on RP18 silica. The fractions containing the product were freeze-dried to give 10.3 mg (10.5 μmol, 8%, 96.5% purity by HPLC Method B) of a white solid. LCMS (Method A) retention time 2.86 min, [M+H] + =868.2. [Example]

[0472] Synthesis scheme of compound XXXI [ka]

[0473] Preparation of methyl (E)-5-bromo-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)nicotinate (XX7.1) [ka]

[0474] Methyl 5-bromo-6-chloronicotinate (15.0 g, 59.8 mmol) and (E)-tert-butyl (4-aminobut-2-en-1-yl)carbamate (13.4 g, 71.8 mmol) were dissolved in DMSO (200 mL). DIPEA (42.1 mL, 239 mmol) was added, and the reaction mixture was stirred at room temperature for 67 hours. (E)-tert-butyl (4-aminobut-2-en-1-yl)carbamate (1.04 g, 5.98 mmol) was added again, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ice water (200 mL) and extracted with ethyl acetate (2 x 250 mL). The organic layer was dried over Na2SO4 and filtered. The solvent was removed under reduced pressure. The residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 200 g, 200 mL / min, 0-80% ethyl acetate in n-hexane over 30 min) to give 18.1 g (45.2 mmol, 75%) of a colorless oil. LCMS (Method A) retention time 2.82 min, [M+H] + =399.9. 1 H NMR(300MHz,DMSO-d6)δ[ppm]=8.59~8.51(m,1H), 8.12~8.04(m,1H), 7.30(t,J=5.8Hz,1H), 6.90(t, J=6.0Hz,1H), 5.69~5.43(m,2H), 4.11~3.96(m,2H), 3.78(s,3H), 3.51(t,J=5.0Hz,2H), 1.35(s,9H).

[0475] Preparation of methyl (E)-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-((3-methoxy-3-oxopropyl)thio)nicotinate (XX7.2) [ka]

[0476] Methyl (E)-5-bromo-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)nicotinate (18.1 g, 45.2 mmol) was coevaporated with anhydrous 1,4-dioxane (30 mL). The residue was dissolved in anhydrous 1,4-dioxane (200 mL), and DIPEA (15.5 mL, 90.4 mmol) and methyl 3-mercaptopropanoate (15 mL, 0.14 mol) were added. Xantphos (2.6 g, 4.5 mmol) and Pd2(dba)3 (1.0 g, 2.2 mmol) were then added, and the resulting mixture was heated to 110 °C for 1.5 h. Water was added, and the aqueous layer was extracted with ethyl acetate. The organic layer was concentrated under reduced pressure (to 150 mL) and filtered through a small pad of Celite® Hyflo Supercel. The solvent was removed under reduced pressure. The residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 200 g, 200 mL / min, 0-100% ethyl acetate in n-hexane over 30 min) to give 17.0 g (38.6 mmol, 85%) of a colorless oil. LCMS (Method A) retention time 3.67 min, [M+H] + =440.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.58~8.52(m,1H), 7.98~7.91(m,1H), 7.27(t,J=5.9Hz,1H), 6.89(t,J=5.8Hz,1H), 5.70~5.43(m, 2H), 4.09~3.96(m,2H), 3.78(s,3H), 3.57(s,3H), 3.49(d,J=5.8Hz,2H), 2.97(t,J=6.9Hz,2H), 2.56(t,J=6.9Hz,2H), 1.35(s,9H).

[0477] Preparation of sodium (E)-2-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-(methoxycarbonyl)pyridine-3-thiolate (XX7.3) [ka]

[0478] Compound XX7.2 (6.0 g, 13 mmol) was dissolved in THF (40 mL). Sodium methylate (3.1 mL, 13 mmol) was then added, and the mixture was stirred at room temperature for 30 minutes. The mixture was diluted with CHCl. ​​The resulting solid was filtered off, washed with CHCl, and dried in vacuo to give 4.5 g (12 mmol, 88%) of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 1.73 min, [M+H] + =354.0. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=7.96~7.89(m,1H), 7.58~7.51(m,1H), 7.38(t,1H), 6.95( t,1H), 5.78~5.48(m,2H), 4.01~3.91(m,2H), 3.70(s,3H), 3.61~3.47(m,2H), 1.37(s,9H).

[0479] Preparation of methyl (E)-3-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-imino-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (XX7.4) [ka]

[0480] Compound XX7.3 (4.5 g, 12 mmol) was dissolved in CHOH (50 mL) and cooled to 0 °C. BrCN (2.1 g, 20 mmol) was then added, and the reaction was stirred at room temperature for 15 minutes. The solvent was removed under reduced pressure. The residue was dissolved in CHCl and washed with water. The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure, and the residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 50 g, 120 mL / min, 0-100% ethyl acetate in petroleum ether over 30 minutes) to give 2.7 g (7.1 mmol, 60%) of a yellow solid. LCMS (Method A) retention time 2.13 min, [M+H] + =379.0. 1H-NMR(300-MHz,DMSO-d6)δppm=8.99(s,1H), 8.67~8.58(m,1H), 8.30~8.24(m,1H), 6.89(t,J=6.1 Hz,1H), 5.70~5.48(m,2H), 4.55(d,J=4.5Hz,2H), 3.84(s,3H), 3.49(t,J=5.0Hz,2H), 1.34(s,9H).

[0481] Preparation of methyl (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (XX7.5) [ka]

[0482] Compound XX7.4 (1.20 g, 3.17 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (586 mg, 3.81 mmol) were dissolved in anhydrous DMF (10 mL). HATU (1.8 g, 4.7 mmol) and DIPEA (2.8 mL, 16 mmol) were then added, and the reaction mixture was stirred at room temperature for 15 minutes. The mixture was quenched with water (20 mL), and the solid was filtered off and washed with water. The product was dried in vacuo to give 1.55 g (3.01 mmol, 95%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 2.88 min, [M+H] + =515.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=9.10~9.03(m,1H), 8.56~8.48(m,1H), 6.87~6.80(m,1H), 5.93~5.84(m,2H), 5.20~5.12 (m,2H), 4.69(q,J=7.1Hz,2H), 4.54(s,1H), 3.98(s,3H), 3.81~3.67(m,2H), 2.32(s,3H), 1.51~1.41(m,3H), 1.39(s,9H).

[0483] Preparation of (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylic acid (XX7.6) [ka]

[0484] Compound XX7.5 (1.5 g, 2.9 mmol) was dissolved in CHOH / THF / HO (2:2:1, 25 mL). LiOH (209 mg, 8.74 mmol) was then added, and the resulting reaction mixture was stirred at room temperature for 2.5 h. The organic solvent was removed under reduced pressure. The aqueous phase was cooled to 0 °C and neutralized with 1 M aqueous HCl. The precipitate was filtered and washed with water. The solid was dissolved / suspended in CHCN / HO (1:1, 15 mL) and freeze-dried to give 1.18 g (2.36 mmol, 81%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 2.49 min, [M+H] + =501.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=13.42(brs,1H), 9.01~8.94(m,1H), 8.90~8.82(m,1H), 6.90(t,J=5.8Hz,1H), 6.84~6.78(m,1H), 5. 80~5.70(m,2H), 5.08(d,J=4.5Hz,2H), 4.57(q,J=7.1Hz,2H), 3.50(t,J=4.8Hz,2H), 2.20(s,3H), 1.35(t,J=7.1Hz,3H), 1.30(s,9H).

[0485] Preparation of tert-butyl ((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)carbamate (XX7.7) [ka]

[0486] Compound XX7.6 (1.18 g, 2.36 mmol) was dissolved in DMF (8 mL), followed by the addition of NH4Cl (378 mg, 7.07 mmol), HATU (1.34 g, 3.54 mmol), and DIPEA (1.21 mL, 7.07 mmol), followed by stirring at room temperature for 25 minutes. The reaction mixture was diluted with water (30 mL). The precipitate was filtered off and washed with water. The solid was dried in vacuo to give 1.1 g (2.2 mmol, 93%) as an off-white solid, which was used without further purification. LCMS (Method A) retention time 2.23 min, [M+H] + =500.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=9.00~8.94(m,1H), 8.80~8.74(m,1H), 8.18(brs,1H), 7.62(brs,1H), 6.90(t,J=5.9Hz,1H), 6.85~6.79(m, 1H), 5.83~5.65(m,2H), 5.09(d,J=4.5Hz,2H), 4.59(q,J=7.1Hz,2H), 3.51(t,J=4.7Hz,2H), 2.21(s,3H), 1.35(t,J=7.1Hz,3H), 1.30(s,9H).

[0487] Preparation of (Z)-3-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxamide hydrochloride (XX7.8) [ka]

[0488] Compound XX7.7 (1.80 g, 3.60 mmol) was suspended in 4 M HCl in dioxane (20.3 mL, 81.1 mmol). The resulting suspension was stirred at room temperature for 30 minutes. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 1.58 g (3.60 mmol, 100%) as an off-white solid, which was used without further purification. LCMS (Method A) retention time 1.36 min, [M+H] + =400.1. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=9.05~8.96(m,1H), 8.91~8.58(m,1H), 8.30(brs,1H), 8.12(brs,2H), 7.64(brs,1H), 6.89~6.83(m,1H) , 6.17~6.01(m,1H), 5.87~5.70(m,1H), 5.18~5.09(m,2H), 4.58(q,J=7.1Hz,2H), 3.48~3.34(m,2H), 2.21(s,3H), 1.36(t,J=7.1Hz,3H).

[0489] Preparation of tert-butyl 4-(5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)amino)-3-nitrophenoxy)butanoate (XX7.9) [ka]

[0490] Compound XX7.8 (1.55 g, 3.56 mmol) was suspended in n-butanol (25 mL), followed by the addition of DIPEA (3.1 mL, 18 mmol) and sodium bicarbonate (597 mg, 7.11 mmol). The mixture was stirred at room temperature for 10 minutes, followed by the addition of tert-butyl 4-(5-carbamoyl-2-chloro-3-nitrophenoxy)butanoate (1.28 g, 3.56 mmol). The reaction mixture was then stirred at 130 °C for 29 hours. The reaction was cooled to room temperature and quenched with water (50 mL). The aqueous layer was extracted with CHCl / CHOH (9:1, 100 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure and the resulting residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar HC 50 g, 120 mL / min, 0-20% CHOH in CH2Cl2 over 30 min) to give 1.25 g (1.73 mmol, 49%) of an orange solid. LCMS (Method A) retention time 3.61 min, [M+H] + =722.2. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.95~8.87(m,1H), 8.78~8.70(m,1H), 8.16(s,1H), 8.05~7.9 9(m,1H), 7.91(brs,1H), 7.68~7.58(m,2H), 7.43~7.36(m,1H), 7.26(brs,1H), 6.75~6.69(m,1H) ), 5.92~5.76(m,2H), 5.04(d,J=4.6Hz,2H), 4.55(q,J=7.1Hz,2H), 4.09(t,J=5.3Hz,2H), 3.88( t,J=6.3Hz,2H), 2.26(t,J=7.3Hz,2H), 2.17(s,3H), 1.84(p,J=6.8Hz,2H), 1.43~1.25(m,12H).

[0491] Preparation of tert-butyl 4-(3-amino-5-carbamoyl-2-(((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)amino)phenoxy)butanoate (XX7.10) [ka]

[0492] Compound XX7.9 (1.25 g, 1.73 mmol) was dissolved in CHOH (20 mL) and cooled to 0 °C. Sodium dithionite (3.0 g, 17 mmol) in water (5 mL) and aqueous NH (30% aqueous solution, 1.7 mL, 13 mmol) were then added. The reaction was warmed to room temperature and stirred at room temperature for 1 h. The reaction was diluted with HO (75 mL) followed by the addition of CHCl (100 mL). The aqueous layer was extracted with CHCl / CHOH (8:2, 2 × 50 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure to give 1.07 g (1.55 mmol, 89%) of an orange solid, which was used without further purification. LCMS (Method A) retention time 2.02 min, [M+H] + =692.3. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=9.00~8.91(m,1H), 8.80~8.70(m,1H), 8.18(s,1 H), 7.67~7.49(m,2H), 6.98~6.86(m,1H), 6.84~6.78(m,2H), 6.74~6.65(m,1H), 5. 92~5.81(m,2H), 5.10~5.03(m,2H), 4.64~4.51(m,4H), 3.91~3.78(m,3H), 3.58~3. 52(m,2H), 2.34~2.23(m,2H), 2.20(s,3H), 1.88~1.73(m,2H), 1.39~1.26(m,12H).

[0493] Preparation of tert-butyl 4-((2-amino-5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX7.11) [ka]

[0494] Compound XX7.10 (1.07 g, 1.55 mmol) was dissolved in CHOH (20 mL), cooled to 0 °C, and BrCN (327 mg, 3.09 mmol) was added. The reaction mixture was stirred at room temperature for 3.5 h. The solvent was removed under reduced pressure. The residue was suspended in petroleum ether, and the solid was filtered off and washed with petroleum ether. The product was dried in vacuo to give 961 mg (1.34 mmol, 86%) of a yellowish solid, which was used without further purification. LCMS (Method A) retention time 1.76 min, [M+H] + =717.2. 1 H-NMR(300MHz,DMSO-d6)δ[ppm]=8.97~8.91(m,1H), 8.82~8.72(m,1H), 8.53(brs,2H), 8.18(brs, 1H), 8.02(brs,1H), 7.64(brs,1H), 7.52~7.43(m,1H), 7.40(brs,1H), 7.36~7.27(m,1H), 6.64~6. 58(m,1H), 5.91(q,J=2.9Hz,2H), 5.13~5.05(m,2H), 4.89~4.82(m,2H), 4.50(q,J=7.1Hz,2H), 3.9 9(t,J=6.3Hz,2H), 2.21(t,J=7.3Hz,2H), 2.13(s,3H), 1.78(q,J=6.8Hz,2H), 1.43~1.20(m,12H).

[0495] Preparation of tert-butyl 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoate (XX7.12) [ka]

[0496] Compound XX7.11 (900 mg, 1.26 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (193 mg, 1.26 mmol) were dissolved in DMF (10 mL). HATU (723 mg, 1.88 mmol) and DIPEA (331 μL, 1.88 mmol) were then added, and the reaction mixture was stirred at room temperature for 4 hours. HATU (241 mg, 634 μmol) and DIPEA (331 μL, 1.88 mmol) were added again, and the mixture was stirred at room temperature for 14 hours. Product formation was monitored by LC / MS. The mixture was quenched with water (20 mL), and the solid was filtered off and washed with water. The residue was suspended in acetonitrile, and the solid was filtered off and dried in vacuo. The resulting residue was suspended in MeOH, and the solid was filtered off. The product was dried in vacuo to give 327 mg (383 μmol, 31%) of an off-white solid which was used without further purification. LCMS (Method A) retention time 2.39 min, [M+H] + =853.3. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=12.78(br s,1H), 8.95~8.88(m,1H), 8.78~8.71(m,1H), 8.14(brs,1H), 7.92(brs,1H), 7.66~7.58 (m,2H), 7.31~7.25(m,2H), 6.63~6.57(m,1H), 6.51~6.45(m,1H), 6.09~5.73(m,2H), 5. 08(d,J=5.7Hz,2H), 4.93(d,J=5.2Hz,2H), 4.50(p,J=7.4Hz,4H), 4.00(t,J=6.3Hz,2H) , 2.25(t,J=7.2Hz,2H), 2.09(s,6H), 1.88~1.73(m,2H), 1.34(s,9H), 1.30~1.20(m,6H).

[0497] Preparation of 4-((5-carbamoyl-1-((E)-4-((Z)-6-carbamoyl-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1H-benzo[d]imidazol-7-yl)oxy)butanoic acid (XX7.13) [ka]

[0498] Compound XX7.12 (230 mg, 270 μmol) was suspended in dioxane (2 mL) and 4 M HCl in dioxane (2.16 mL, 8.63 mmol) was added. The resulting suspension was stirred at room temperature for 1 hour. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 217 mg (260 μmol, 96%) of an off-white solid, which was used without further purification. LCMS (Method A) retention time 1.84 min, [M+H] + =797.5. 1H-NMR(300MHz,DMSO-d6)δ[ppm]=8.92(d,J=2.0Hz,1H), 8.74(d,J=2.0Hz,1H), 8.16(s,1H), 7 .94(s,1H), 7.66~7.57(m,2H), 7.34~7.28(m,2H), 6.59(s,1H), 6.49(s,1H), 6.07~5.93(m,1H) , 5.89~5.74(m,1H), 5.07(d,J=5.7Hz,2H), 4.95(d,J=5.3Hz,2H), 4.49(p,J=7.2Hz,4H), 4.03( t,J=6.4Hz,2H), 2.28(t,J=7.1Hz,2H), 2.09(s,6H), 1.83(p,J=6.7Hz,2H), 1.32~1.20(m,6H).

[0499] Preparation of (Z)-3-((E)-4-(5-carbamoyl-7-(4-(1,2-dimethylhydrazinyl)-4-oxobutoxy)-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-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxamide (Compound XXXI) [ka]

[0500] Compound XX7.13 (50.0 mg, 60.0 μmol) was suspended in DMF (3 mL). Then, DIPEA (31.3 μL, 180 μmol), NHS (14.1 mg, 120 μmol), and DIC (18.6 μL, 120 μmol) were added, and the resulting reaction mixture was stirred at 40 °C for 6.6 hours. NHS (7.0 mg, 60 μmol) and DIPEA (10 μL, 60 μL) were added again, and the reaction mixture was stirred at 45 °C for 15 hours. The reaction mixture was then stirred at room temperature for 15 hours. The progress of the reaction was monitored by LC / MS. Next, 1,2-dimethylhydrazine dihydrochloride (39.9 mg, 300 μmol) and DIPEA (104 μL, 600 μmol) were added, and the resulting reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by LC / MS. The solvent was removed under reduced pressure and the residue was purified by preparative HPLC (Method I) on RP18 silica. The product-containing fractions were freeze-dried to give 8.0 mg (8.4 μmol, 14%, 95.6% purity by HPLC Method A) of a white solid. LCMS (Method D) retention time 4.70 min, [M+H] + =839.4. [Example]

[0501] Synthesis scheme of compound XXXII [ka]

[0502] Preparation of methyl (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX4.1) [ka]

[0503] Compound XX3.2 (1.00 g, 2.45 mmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (450 mg, 2.95 mmol) were dissolved in DMF (10 mL). HATU (1.41 g, 3.68 mmol) and DIPEA (2.16 mL, 12.3 mmol) were then added, and the reaction mixture was stirred at room temperature for 1 hour. Product formation was monitored by LC / MS. Water was then added, and the resulting solid was filtered off and washed with water. The product was dried in vacuo to give 1.40 g of a yellowish solid, which was used without further purification. LCMS (Method C) retention time 2.91 min, [M+Na]+ = 567.3. 1 H-NMR: (300MHz, CDCl3)δ[ppm]=7.96(d,J=1.4Hz,1H), 7.61(d,J=1.4Hz,1H), 5.91~5.63(m,2H), 5.40(dd,J=5.5,1.3Hz,2H), 4.5 0(brs,1H), 4.02(s,3H), 3.95(s,3H), 3.76~3.66(m,2H), 3.07(q,J=7.6Hz,2H), 2.53(s,3H), 1.39(s,9H), 1.29(t,J=7.6Hz,3H).

[0504] Preparation of methyl (Z)-3-((E)-4-aminobut-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX4.2) [ka]

[0505] Compound XX4.1 (1.40 g, 2.57 mmol) was suspended, and 4 M HCl in dioxane (10.3 mL, 41.1 mmol) was added. The resulting suspension was stirred at room temperature for 1 hour. Product formation was monitored by LC / MS. The formed solid was filtered off, washed with diethyl ether, and dried in vacuo to give 1.07 g (2.22 mmol, 94%) of a yellowish solid. LCMS (Method A) retention time 1.87 min, [M+H]+ = 445.1. 1H-NMR: (300MHz,DMSO-d6)δ[ppm]=8.34~7.93(m,2H), 7.57~7.49(m,1H), 7.12(brs,1H), 6.12~5.96(m,1H), 5.77~5.62(m,1H), 5.31(d,J=5.7Hz,2H), 4.01(s,3H), 3.87(s,3H), 3.47~3.35(m,2H), 2.96(q,J=7.5Hz,2H), 2.47(s,3H), 1.20(t,J=7.5Hz,3H).

[0506] Preparation of methyl (Z)-3-((E)-4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX4.3) [ka]

[0507] Methyl (Z)-3-((E)-4-aminobut-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (100 mg, 208 μmol) was suspended in dioxane (4 mL), 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (71.5 mg, 208 μmol), DIPEA (73.2 μL, 415 μmol), and KCO (86.2 mg, 624 μmol), and the mixture was stirred at 120 °C for 24 h. Again, KCO (57.5 mg, 415 μmol) was added, and the reaction mixture was stirred at 120 °C for 4 days. Then, K2CO3 (57.5 mg, 415 μmol) was added, and the reaction mixture was stirred at 120 °C for 23 h. Again, 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (17.8 mg, 52.0 μmol), DIPEA (36.6 μL, 208 μmol), and K2CO3 (57.5 mg, 415 μmol) were added, and the reaction mixture was stirred at 120 °C for 4 h. The formation of the product was monitored by LC / MS. The reaction was cooled to room temperature and quenched with water (10 mL). The product was extracted with CH2Cl2 / CH3OH (9 / 1, 2 × 20 mL). The organic layer was dried over Na2SO4 and filtered. The solvent was removed under reduced pressure. The residue was purified by automated flash column chromatography using a Buchi C-850 (XSelect® CSH™ Prep C18 0.5 μM, 50 mm × 150 mm, 100 mL / min, 15–100% CHCN in HO (+0.15% TFA) over 30 min). Fractions containing the product were combined to give 25 mg (29 μmol, 14%) of an orange solid. LCMS (Method B) retention time 2.71 min, [M+H] = 752.3. 1H-NMR:(300MHz,DMSO-d6)δ[ppm]=8.16~8.09(m,1H), 8.03~7.97(m,1H), 7.87(brs,1H), 7.61~7.46(m,2 H), 7.44~7.37(m,1H), 7.25(brs,1H), 5.83~5.55(m,2H), 5.27(d,J=5.5Hz,2H), 4.07(d,J=6.3Hz,2H), 4. 01~3.92(m,4H), 3.90(s,3H), 3.85(s,3H), 3.62(t,J=12.1Hz,2H), 3.40(d,J=12.4Hz,2H), 3.29~3.14(m, 2H), 3.06~3.00(m,2H), 2.91(q,J=7.5Hz,2H), 2.48(s,3H), 2.06(d,J=9.5Hz,2H), 1.15(t,J=7.6Hz,3H).

[0508] Preparation of methyl (Z)-3-((E)-4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX4.4) [ka]

[0509] To a stirred solution of compound XX4.3 (1.15 g, 1.33 mmol) in CHOH (20 mL) was added sodium dithionite (2.31 g, 13.3 mmol) dissolved in water (5 mL) at 0 °C. Then, NH (30% aqueous solution, 1.32 mL, 9.96 mmol) was added to the mixture at 0 °C. The reaction was allowed to warm to room temperature and stirred at room temperature for 30 min. The progress of the reaction was monitored by LC / MS. The reaction mixture was diluted with H O (75 mL) and CHCl (100 mL) was added. The aqueous layer was extracted with ethyl acetate (2 × 50 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure and dried in vacuo to give 845 mg (1.17 mmol, 88%) of a yellowish solid. The residue was used without further purification. LCMS (Method A) retention time 1.76 min, [M+H] + =722.3. 1 H-NMR:(300MHz,DMSO-d6)δ[ppm]=8.16~8.08(m,1H), 7.59~7.42(m,2H), 6.92(s,1H), 6.84 ~6.76(m,1H), 6.66~6.60(m,1H), 5.87~5.57(m,2H), 5.24(d,J=5.4Hz,2H), 4.60(d,2H), 3.8 8(d,J=2.8Hz,7H), 3.74(t,J=6.3Hz,2H), 3.63~3.53(m,2H), 3.49(t,J=4.6Hz,4H), 2.94(q, J=7.5Hz,2H), 2.46(s,3H), 2.29~2.17(m,6H), 1.65(p,J=6.6Hz,2H), 1.16(t,J=7.5Hz,3H).

[0510] Preparation of methyl (Z)-3-((E)-4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate trifluoroacetate (XX4.5) [ka]

[0511] Compound XX4.4 (250 mg, 346 μmol) was dissolved in anhydrous DMF (4 mL) and cooled to 0° C. Then, 4-ethyl-2-methyloxazole-5-carbonyl isothiocyanate (0.2 M in dioxane, 865 μL, 173 μmol) was added dropwise, and the reaction mixture was stirred at 0° C. for 30 minutes. Again, 4-ethyl-2-methyloxazole-5-carbonyl isothiocyanate (0.2 M in dioxane, 430 μL, 87.0 μmol) was added dropwise, and the reaction mixture was stirred at 0° C. for 15 minutes. Then, 4-ethyl-2-methyloxazole-5-carbonyl isothiocyanate (0.2 M in dioxane, 430 μL, 87.0 μmol) was added dropwise, and the reaction mixture was stirred at 0° C. for 1 hour. EDC·HCl (83.0 mg, 433 μmol) and EtN (121 μL, 866 μmol) were then added, and the reaction mixture was stirred at room temperature for 2 days. Product formation was monitored by LC / MS. The reaction mixture was diluted with water / saturated aqueous NH4Cl (3:1, 20 mL) and extracted with CHCl2 / CH3OH (3:1, 3 × 25 mL). The combined organic layers were dried over sodium sulfate and filtered. The solvent was removed under reduced pressure. The residue was purified by flash column chromatography on a Buchi C-850 (XSelect® CSH™ Prep OBD™ C18 0.5 μM, 50 mm × 150 mm, 100 mL / min, 5–100% CH3CN in HO (+0.15% TFA) for 30 min) to give 240 mg (240 μmol, 69%) of an off-white solid. LCMS (Method C) retention time 3.31 min, [M+H] + =884.4. 1H-NMR:(300MHz,DMSO-d6)δ[ppm]=9.91(s,1H), 8.15~8.09(m,1H), 7.90(s,1H), 7.68~7.62(m,1H), 7.47~ 7.40(m,1H), 7.34(s,1H), 7.29~7.22(m,1H), 5.83~5.60(m,2H), 5.26(d,J=4.6Hz,2H), 4.88(d,J=4.5Hz,2 H), 4.00(t,J=5.8Hz,2H), 3.94~3.84(m,5H), 3.72(s,3H), 3.67~3.55(m,2H), 3.38~3.24(m,2H), 3.22~3. 10(m,2H), 3.03~2.97(m,2H), 2.86~2.71(m,4H), 2.46~2.35(m,6H), 1.98~1.87(m,2H), 1.09~0.96(m,6H).

[0512] Preparation of (Z)-3-((E)-4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylic acid (XX4.6) [ka]

[0513] Compound XX4.5 (100 mg, 100 μmol) was dissolved in CHOH / THF / HO (2:2:1, 2.5 mL). LiOH (25.2 mg, 601 μmol) was then added, and the resulting reaction mixture was stirred at room temperature for 18 h. The reaction progress was monitored by LC / MS. The organic solvent was removed under reduced pressure. The aqueous phase was cooled to 0 °C and neutralized with 1 M aqueous HCl. The precipitate was filtered and washed with water. The solid was dissolved / suspended in CHCN / HO (1:1, 15 mL) and freeze-dried to give 64 mg (74 μmol, 73%) of a white solid. The product was used without further purification. LCMS (Method A) retention time 1.86 min, [M+H] + =870.5. 1 H-NMR:(300MHz,DMSO-d6)δ[ppm]=13.16(brs,1H), 12.73(brs,1H), 10.94(brs,1H), 8.14~8.08(m,1H), 7 .93(s,1H), 7.68~7.62(m,1H), 7.55~7.48(m,1H), 7.35~7.28(m,1H), 5.89~5.63(m,2H), 5.28(d,J=4.9Hz, 2H), 4.90(d,J=4.8Hz,2H), 4.11~4.01(m,2H), 3.92~3.86(m,2H), 3.82~3.64(m,5H), 3.16~3.10(m,2H), 3. 02~2.96(m,2H), 2.85~2.71(m,4H), 2.47~2.33(m,6H), 2.10~1.92(m,2H), 1.24(s,2H), 1.07~0.93(m,6H).

[0514] Preparation of Compound XXXII [ka]

[0515] Compound XX4.6 (62.0 mg, 71.3 μmol) was suspended in DMF (3 mL). Then, DIPEA (37.2 μL, 214 μmol), NHS (16.7 mg, 143 μmol), and DIC (22.1 μL, 143 μmol) were added, and the resulting reaction mixture was stirred at 40 °C for 3 h. The reaction was then stirred at room temperature for 17 h. The reaction progress was monitored by LC / MS. Next, 1 M hydrazine in THF (356 μL, 356 μmol) and DIPEA (62 μL, 356 μmol) were added, and the resulting reaction mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LC / MS. The solvent was removed under reduced pressure, and the residue was purified by preparative HPLC (Method I) on RP18 silica. The product-containing fractions were freeze-dried to give 43.0 mg (43.1 μmol, 60%, 95.9% purity by HPLC Method A) of a white solid. LCMS (Method C) retention time 2.72 min, [M+H] + =884.4. [Example]

[0516] Synthesis scheme of compound XXXIII [ka]

[0517] Preparation of tert-butyl (E)-(4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate trifluoroacetate (XX2.1) [ka]

[0518] Compound XX.3 (1.96 g, 4.01 mmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (685.7 mg, 4.41 mmol) were dissolved in DMF (20 mL) and then treated with DIPEA (2.65 mL, 16.1 mmol) at 0 °C for 15 min. HATU (1.98 g, 5.22 mmol) was then added, and the resulting yellowish reaction mixture was stirred at room temperature for 1 h. Product formation was monitored by LCMS. The solvent was removed in vacuo, and the residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 250 mm, 100 mL / min, 10–100% CH3CN in HO (+0.15% TFA) for 47 min). The product-containing fractions were combined and freeze-dried to give 1.87 g (2.53 mmol, 63%) of a reddish solid. LCMS (Method C) retention time 2.57 min, [M+H] + =626.2. 1H-NMR (300MHz, DMSO-d6): d[ppm]=9.80(brs,1H), 7.95(brs,1H), 7.68(dd,J=2.5,1.2Hz,1H), 7.4 2~7.31(m,2H), 6.97~6.85(m,1H), 5.84~5.67(m,1H), 5.61~5.39(m,1H), 5.03~4.90(m,2H), 4.26( t,J=5.8Hz,2H), 4.03(d,J=12.6Hz,2H), 3.68(t,J=12.1Hz,2H), 3.60~3.40(m,4H), 3.37~3.26(m, 2H), 3.25~3.06(m,2H), 2.99(q,J=7.5Hz,2H), 2.44(s,3H), 2.31~2.15(m,2H), 1.39~1.06(m,12H).

[0519] Preparation of (E)-N-(1-(4-aminobut-2-en-1-yl)-5-carbamoyl-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methyloxazole-5-carboxamide hydrochloride (XX2.2) [ka]

[0520] Compound XX2.1 (1.87 g, 2.53 mmol) was dissolved / suspended in CHCl (10 mL). HCl (4 M in dioxane, 5.06 mL) was then added, and the resulting mixture was stirred at room temperature for 24 h. The reaction progress was monitored by LCMS. HCl (0.15% in HO) was added, and then all volatiles were removed in vacuo. The residue was purified on RP18 silica by automated flash column chromatography on a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CHCN in HO (+0.15% HCl) over 47 min). The product-containing fractions were combined and freeze-dried to give 1.18 g (2.10 mmol, 83%) of a white solid. LCMS (Method C) retention time 1.67 min, [M+H] + =526.1. 1H-NMR (300MHz, DMSO-d6): d[ppm]=11.65(s,1H), 8.28~8.22(m,3H), 7.70(d,J =1.2Hz,1H), 7.52~7.33(m,2H), 6.16~6.06(m,1H), 5.70~5.50(m,1H), 4.99(d ,J=5.2Hz,2H), 4.30(t,J=6.0Hz,2H), 4.05~3.84(m,4H), 3.57~3.39(m,4H), 3 .36~3.08(m,4H), 2.99(q,J=7.5Hz,2H), 2.50~2.30(m,5H), 1.29~1.15(m,3H).

[0521] Preparation of methyl (E)-5-bromo-6-((4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)nicotinate trifluoroacetate (XX2.3) [ka]

[0522] Compound XX2.2 (1.07 g, 1.90 mmol) and methyl 5-bromo-6-chloronicotinate (572 mg, 2.28 mmol) were dissolved in DMF (25 mL). EtN (2.65 mL, 19.0 mmol) was then added, and the resulting reaction mixture was heated to 55 °C for 3 days. Product formation was monitored by LCMS. All volatiles were removed in vacuo, and the residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 5–100% CH3CN in HO (+0.15% TFA) for 40 min). The product-containing fractions were freeze-dried to yield 759 mg (890 μmol, 47%) of a white solid. LCMS (Method C) retention time 1.63 min, [M+H] + =739.1. 1H-NMR (300MHz, DMSO-d6): d[ppm]=12.75(brs,1H), 10.10(brs,1H), 8.47(d,J=2.0Hz,1H), 8.07(d,J=2.0Hz,1H ), 7.94(brs,1H), 7.67(d,J=1.2Hz,1H), 7.39~7.25(m,3H), 5.88~5.73(m,1H), 5.73~5.58(m,1H), 4.92(d,J=5.3 Hz,2H), 4.21(t,J=5.8Hz,2H), 4.06~3.96(m,4H), 3.79(s,3H), 3.70(d,J=12.0Hz,1H), 3.47(d,J=12.1Hz,2H), 3 .29(t,J=8.1Hz,2H), 3.12(s,2H), 2.89(q,J=7.5Hz,2H), 2.42(s,3H), 2.22~2.10(m,2H), 1.09(t,J=7.5Hz,3H).

[0523] Preparation of methyl (E)-6-((4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)-5-((3-methoxy-3-oxopropyl)thio)nicotinate trifluoroacetate (XX2.4) [ka]

[0524] Compound XX2.3 (750 mg, 879 μmol) was coevaporated with anhydrous 1,4-dioxane (7.5 mL). The residue was dissolved in anhydrous 1,4-dioxane (7.5 mL), and DIPEA (451 μL, 2.65 mmol) and methyl 3-mercaptopropanoate (389 μL, 3.51 mmol) were added. Xantphos (50.8 mg, 87.9 μmol) and Pd2(dba)3 (40.2 mg, 43.9 μmol) were then added under a slight argon flow, and the resulting mixture was heated to 130 °C for 30 min. Product formation was monitored by LCMS. Water was added, and then all solvents were removed in vacuo. The residue (filtered through syringe filter PVDF-45 / 25) was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CH3CN in HO (+0.15% TFA) over 47 min). Fractions containing the product were freeze-dried to give 682 mg (764 μmol, 87%) of a colorless solid. LCMS (Method C) retention time 2.28 min, [M+H] + =779.2. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=12.77(brs,1H), 10.01(brs,1H), 8.48(d,J=2.2Hz,1H), 7.97~7.88(m,2H), 7.67(d,J=1.3Hz,1H), 7.39~7.33(m,2H), 7.26(t,J=5.9Hz,1H), 5.88~5.58(m,2H), 4.93(d,J=5.3Hz,2H), 4.2 0(t,J=5.8Hz,2H), 4.09~3.96(m,4H), 3.79(s,3H), 3.68(t,J=12.0Hz,2H), 3.51(s,3H), 3.45(s,1H), 3.37~3. 23(m,2H), 3.15~3.07(m,2H), 2.95~2.81(m,4H), 2.48~2.39(m,5H), 2.22~2.10(m,2H), 1.08(t,J=7.5Hz,3H).

[0525] Preparation of methyl (E) 6-((4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)-5-mercaptonicotinate trifluoroacetate (XX2.5) [ka]

[0526] Compound XX2.4 (670 mg, 750 μmol) was dissolved in anhydrous THF (7.5 mL), and 25% NaOCH in CHOH (686 μL, 3.00 mmol) was added under a slight argon stream. The resulting mixture was stirred for 2 h. Product formation was monitored by LCMS. The mixture was diluted with CHCl (5 mL), and the resulting solid was filtered and washed with CHCl (3 × 2 mL). The residue was purified on RP18 silica by automated flash column chromatography using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CHCN in HO (+0.15% TFA) for 43 min). The product-containing fractions were freeze-dried to yield 396 mg (491 μmol, 65%) of a yellowish solid that is prone to disulfide bond formation. LCMS (Method C) retention time 1.87 min, [M+H] + =693.2.

[0527] Preparation of methyl (E)-3-(4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-imino-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (XX2.6) [ka]

[0528] Compound XX2.5 (390 mg, 483 μmol) was dissolved in CHOH (7.5 mL) and cooled to 0 °C. Cyanogen bromide (61.4 mg, 580 μmol) was then added, and the mixture was stirred at 0 °C for 1 h. The mixture was then warmed to room temperature and stirred for 1 h. 0.5 M aqueous TCEP solution (1 mL) was then added, and the mixture was stirred at room temperature for 1 h. 0.5 M aqueous TCEP solution (2 mL) was then added, and the mixture was stirred at room temperature for 2 h. All volatiles were removed in vacuo, and the residue was dissolved in CHOH (7.5 mL) and stirred at room temperature for 1 h. The progress of the reaction was monitored by LCMS. The residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CH3CN in HO (50 mM NH4HCO3) over 47 min). The product was obtained as a mixture with a urea by-product. This material (153 mg) was used in further reactions. LCMS (Method C) retention time 1.62 min, [M+H] + =718.2.

[0529] Preparation of methyl (E)-3-((E)-4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate trifluoroacetate (XX2.7) [ka]

[0530] Compound XX2.6 (153 mg, 213 μmol) and 4-ethyl-2-methyl-1,3-oxazole-5-carboxylic acid (36.4 mg, 234 μmol) were dissolved in DMF (4 mL) and then treated with DIPEA (141 μL, 853 μmol) at 0 °C for 15 min. HATU (105 mg, 277 μmol) was then added, and the resulting yellowish reaction mixture was stirred at room temperature for 2 h. Product formation was monitored by LCMS. The solvent was removed in vacuo, and the residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 5–100% CH3CN in HO (+0.15% TFA) for 40 min). The product-containing fractions were freeze-dried (CHCN / H0=1:1) to give 116 mg (119 μmol, 56%) of a white solid. LCMS (Method C) retention time 3.22 min, [M+H] + =855.2. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=12.73(brs,1H), 9.84(brs,1H), 8.91(dd,J=12.6,2.0Hz,2H), 7.91(brs,1H) ), 7.64(d,J=1.2Hz,1H), 7.37~7.27(m,2H), 5.97~5.83(m,1H), 5.77~5.62(m,1H), 5.02(d,J=5.4Hz,2H), 4.93 ~4.85(m,2H), 4.11(t,J=5.8Hz,2H), 4.02~3.84(m,5H), 3.64(t,J=11.1Hz,2H), 3.47~3.29(m,2H), 3.29~3.15 (m,2H), 3.15~2.96(m,2H), 2.83~2.66(m,4H), 2.43(s,3H), 2.39(s,3H), 2.07~1.98(m,2H), 1.03~0.91(m,5H).

[0531] Preparation of (E)-3-((E)-4-(5-carbamoyl-2-(4-ethyl-2-methyloxazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylic acid (XX2.8) [ka]

[0532] Compound XX2.7 (110 mg, 114 μmol) was dissolved in CH3OH / THF / H2O (2:2:1, 5 mL). LiOH (8.16 mg, 341 μmol) was then added, and the resulting reaction mixture was stirred at room temperature for 3 h. The reaction progress was monitored by LCMS. The organic solvent was removed in vacuo (rotary evaporator). The aqueous phase (approximately 5 mL of additional water was added) was cooled to 0 °C and neutralized with 1 M aqueous HCl. The suspension was transferred to a 50 mL Falcon tube and centrifuged at 7000 rpm for 10 min at 0 °C. The pellet was washed with water (15 mL) and centrifuged at 7000 rpm for 10 min at 0 °C. The solid was dissolved / suspended in CH3CN / H2O (1:1, 25 mL) and freeze-dried. The product (94.9 mg) was used without further purification. LCMS (Method C) retention time 2.89 min, [M+H] + =841.1. 1H-NMR (300MHz, DMSO-d6): d[ppm]=13.39(s,1H), 12.70(s,1H), 11.37(s,1H), 8.94(d,J=2.0Hz,1H ), 8.85(d,J=1.9Hz,1H), 7.94(s,1H), 7.67~7.61(m,1H), 7.41~7.31(m,2H), 5.98~5.83(m,1H), 5.7 8~5.61(m,1H), 5.02(d,J=5.3Hz,2H), 4.89(d,J=4.9Hz,2H), 4.13(t,J=5.8Hz,2H), 4.01~3.74(m,5 H), 3.28~2.96(m,4H), 2.82~2.64(m,4H), 2.45~2.35(m,6H), 2.16~2.05(m,2H), 1.07~0.88(m,6H).

[0533] Preparation of N-(5-carbamoyl-1-((E)-4-((E)-2-((4-ethyl-2-methyloxazole-5-carbonyl)imino)-6-(hydrazinecarbonyl)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-2-yl)-4-ethyl-2-methyloxazole-5-carboxamide (compound XXXIII) [ka]

[0534] Compound XX2.8 (95 mg, 0.13 mmol) was suspended in DMF (3 mL). Then, DIPEA (59 μL, 0.34 mmol), NHS (26 mg, 0.23 mmol), and DIC (35 μL, 0.23 mmol) were added, and the resulting reaction mixture was stirred at room temperature for 5.5 h. The reaction progress was monitored by LC / MS. Then, 1 M hydrazine in THF (564 μL, 564 μmol) and DIPEA (59 μL, 0.34 mmol) were added, and the resulting reaction mixture was stirred at room temperature for 30 min. The reaction progress was monitored by LC / MS. The solvent was removed in vacuo (oil pump vacuum) and freeze-dried (CHCN / HO 1:1, 10 mL). The residue was purified by preparative HPLC (Method III) on RP18 silica. The fractions containing the product were freeze-dried to give 52 mg (54%, 96% purity by HPLC Method A) of a white solid. LCMS (Method B) retention time 1.97 min, [M+H] + =855.2. [Example]

[0535] Synthesis scheme of compound XXXIV [ka]

[0536] Preparation of methyl (E)-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-3-methoxybenzoate (XX3.1) [ka]

[0537] Methyl 4-iodo-3-methoxybenzoate (1.00 g, 3.42 mmol), cesium carbonate (2.04 g, 6.16 mmol), palladium(II) acetate (20.2 mg, 171 μmol), and BINAP (213 mg, 342 μmol) were dissolved in toluene (10 mL). Then, (E)-tert-butyl (4-aminobut-2-en-1-yl)carbamate (957 mg, 5.14 mmol) was added under a slight argon flow, and the resulting mixture was heated to 120 °C for 4 h. The mixture was diluted with water (50 mL), and the product was extracted with DCM (2 × 50 mL). The organic layers were combined, dried over Na SO , filtered, and evaporated. The residue was purified by column chromatography using DCM / [DCM:MeOH (9:1)] (100:00 → 50:50, 120 mL / min, 35 min, Biotage SpA HC 100 g) (linear gradient) as eluent. The product-containing fractions were combined, evaporated, and dried under high vacuum to give 992 mg (2.83 mmol, 83%) of a light brown solid. LCMS (Method B) retention time 3.60 min, [M+H] + =351.2. 1 H-NMR:(300MHz,DMSO-d6)d 7.50~7.41(m,1H), 7.28(d,J=1.8Hz,1H), 6.91(t,J=5.9Hz,1H), 6.51(d,J=8.4Hz,1H), 5.91(t,J= 6.0Hz,1H), 5.59~5.51(m,2H), 3.83(s,3H,CH3), 3.76(s,5H), 3.55~3.46(m,2H), 1.35(s,9H,Boc).

[0538] Preparation of methyl (E)-3-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-imino-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX3.2) [ka]

[0539] Methyl (E)-4-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-3-methoxybenzoate (1.90 g, 5.42 mmol) was dissolved in acetic acid (8 mL), and KSCN (2.11 g, 21.7 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. Then, Br2 (277 μL, 5.42 mmol) dissolved in acetic acid (2 mL) was added to the mixture, and the reaction was stirred at room temperature for 3 hours. The formation of the product was controlled by LC / MS. The mixture was quenched with water (50 mL). The solid was filtered off. The filtrate was adjusted to pH 9 with aqueous ammonia (33%), and the product was extracted with ethyl acetate (2 × 80 mL). The combined organic layers were dried over sodium sulfate and filtered. The solvent was removed under reduced pressure. The residue was purified by automated flash column chromatography on silica using a Biotage Selekt (Biotage Sfar HC 50 g, 120 mL / min, 0–20% CHOH in CH2Cl2 over 30 min) to give 407 mg (982 μmol, 18%) of a white solid. LCMS (Method A) retention time 1.56 min, [M+H]+ = 408.2. 1 H-NMR: (300MHz,DMSO-d6)δ[ppm]=8.51(brs,1H), 7.70~7.62(m,1H), 7.43~7.35(m,1H), 6.90(brs,1H), 5.68~5.44(m,2H), 4.79(d,J=5.1Hz,2H), 3.87(s,3H), 3.82(s,3H), 3.47(d,J=5.6Hz,2H), 1.34(s,9H).

[0540] Preparation of methyl (Z)-3-((E)-4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX3.3) [ka]

[0541] Compound XX3.2 (1.00 g, 2.54 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (450 mg, 2.95 mmol) were dissolved in DMF (10 mL). HATU (1.41 g, 3.68 mmol) and DIPEA (2.16 mL, 12.3 mmol) were then added, and the reaction mixture was stirred at room temperature for 30 minutes. Product formation was monitored by LC / MS. Water was then added, and the resulting solid was filtered off and washed with water. The product was dried in vacuo to give 1.60 g of a yellowish solid, which was used without further purification. LCMS (Method C) retention time 3.13 min, [M+Na]+ = 408.2. 1 H-NMR:(300MHz,CDCl3)δ[ppm]=8.04~7.91(m,1H), 7.64~7.58(m,1H), 6.83~6.76(m,1H), 5.93~5.67(m,2H), 5.45~5.37(m,2H), 4. 70(q,J=7.1Hz,2H), 4.53(brs,1H), 4.02(s,3H), 3.95(s,3H,), 3.77~3.67(m,2H), 2.31(s,3H), 1.47(t,J=7.1Hz,3H), 1.39(s,9H).

[0542] Preparation of methyl (Z)-3-((E)-4-aminobut-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate hydrochloride (XX3.4) [ka]

[0543] Compound XX3.3 (1.60 g, 2.94 mmol) was treated with 4 M HCl in dioxane (11.8 mL, 47.1 mmol). The resulting suspension was stirred at room temperature for 30 minutes. Product formation was monitored by LC / MS. The solid formed was filtered off, washed with diethyl ether, and dried in vacuo to give 996 mg (2.08 mmol, 76%) of a yellowish solid. LCMS (Method A) retention time 1.87 min, [M+H]+ = 444.3. 1H-NMR:(300MHz,DMSO-d6)δ[ppm]=8.22~8.08(m,3H), 7.60~7.53(m,1H), 6.84~6.75(m,1H), 6.14~5.98(m,1H), 5.83~5.68(m,1H) ), 5.35(d,J=5.8Hz,2H), 4.56(q,J=7.1Hz,2H), 4.02(s,3H), 3.88(s,3H), 3.48~3.37(m,2H), 2.20(s,3H), 1.34(t,J=7.1Hz,3H).

[0544] Preparation of methyl (Z)-3-((E)-4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate trifluoroacetate (XX3.5) [ka]

[0545] Compound XX3.4 (100 mg, 208 μmol) was suspended in dioxane (4 mL), 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (71.5 mg, 208 μmol), DIPEA (73.2 μL, 415 μmol), and KCO (86.2 mg, 624 μmol), and the mixture was stirred at 120° C. for 24 hours. Again, KCO (57.5 mg, 415 μmol) was added, and the reaction mixture was stirred at 120° C. for 4 days. Then, KCO (57.5 mg, 415 μmol) was added, and the reaction mixture was stirred at 120° C. for 23 hours. Again, 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (17.8 mg, 52.0 μmol), DIPEA (36.6 μL, 208 μmol), and K2CO3 (57.5 mg, 415 μmol) were added, and the reaction mixture was stirred at 120 °C for 4 h. Product formation was monitored by LC / MS. The reaction was cooled to room temperature and quenched with water (10 mL). The product was extracted with CH2Cl2 / CH3OH (9 / 1, 2 × 20 mL). The organic layer was dried over Na2SO4 and filtered. The solvent was removed under reduced pressure. The residue was purified by automated flash column chromatography using a Buchi C-850 (XSelect® CSH™ Prep OBD™ C18 0.5 μM, 50 mm × 150 mm, 100 mL / min, 15–100% CH3CN in HO (+0.15% TFA) over 30 min). Fractions containing the product were combined and freeze-dried to give 84 mg (97 μmol, 47%) of a yellowish solid. LCMS (Method B) retention time 2.84 min, [M+H]+ = 751.3. 1H-NMR:(300MHz,DMSO-d6)δ[ppm]=8.15~8.08(m,1H), 8.04~7.97(m,1H), 7.88(s,1H), 7.57(s,1H), 7.52~7. 46(m,1H), 7.46~7.36(m,1H), 7.24(s,1H), 6.77~6.70(m,1H), 5.84~5.61(m,2H), 5.29(d,J=5.0Hz,2H), 4.5 6(q,J=7.1Hz,2H), 4.13~4.07(m,2H), 4.00~3.91(m,4H), 3.90(s,3H), 3.85(s,3H), 3.71~3.53(m,2H), 3.48 ~3.30(m,2H), 3.27~3.13(m,2H), 3.10~2.90(m,2H), 2.19(s,3H), 2.12~1.99(m,2H), 1.33(t,J=7.1Hz,3H).

[0546] Preparation of methyl (Z)-3-((E)-4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate (XX3.6) [ka]

[0547] To a stirred solution of compound XX3.5 (1.73 g, 2.30 mmol) in CHOH (30 mL) was added sodium dithionite (4.01 g, 23.0 mmol) dissolved in water (10 mL) at 0 °C. Then, NH (30% aqueous solution, 2.29 mL, 17.3 mmol) was added to the mixture at 0 °C. The reaction mixture was warmed to room temperature and stirred at room temperature for 1.5 h. The progress of the reaction was monitored by LC / MS. The reaction was diluted with HO (75 mL) and CHCl (100 mL) was added. The aqueous layer was extracted with CHCl / CHOH (8:2, 5 × 50 mL). The organic layer was dried over sodium sulfate and filtered. The solvent was removed under reduced pressure to give 1.31 g (1.82 mmol, 79%). The residue was used without further purification. LCMS (Method A) retention time 1.85 min, [M+H] + =721.4. 1 H-NMR:(300MHz,DMSO-d6)δ[ppm]=8.16~8.09(m,1H), 7.57~7.47(m,2H), 6.92(brs,1H), 6 .82~6.77(m,1H), 6.76~6.71(m,1H), 6.67~6.60(m,1H), 5.89~5.64(m,2H), 5.29(d,J=5.2 Hz,2H), 4.65~4.50(m,4H), 3.99~3.83(m,7H), 3.74(t,J=6.2Hz,2H), 3.60(t,J=5.9Hz,2H) ), 3.48(t,J=4.6Hz,4H), 2.28~2.16(m,9H), 1.65(p,J=6.6Hz,2H), 1.33(t,J=7.1Hz,3H).

[0548] Preparation of methyl (Z)-3-((E)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylate trifluoroacetate (XX3.7) [ka]

[0549] Compound XX3.6 (250 mg, 347 μmol) was dissolved in anhydrous DMF (4 mL) and cooled to 0 °C. Then, 1-ethyl-3-methyl-1-pyrazole-5-carbonyl isothiocyanate (0.2 M in dioxane, 867 μL, 173 μmol) was added dropwise, and the reaction was stirred at 0 °C for 1.5 h. Again, 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.2 M in dioxane, 867 μL, 173 μmol) was added, and the mixture was stirred at 0 °C for 30 min. Then, EDC·HCl (83.1 mg, 434 μmol) and EtN (122 μL, 867 μmol) were added, and the reaction was stirred at room temperature for 18 h. EDC·HCl (16.6 mg, 87.0 μmol) and EtN (24.3 μL, 173 μmol) were added again, and the reaction was stirred at room temperature for 72 h. Product formation was monitored by LC / MS. The reaction was diluted with water / saturated aqueous NH₄Cl (3:1, 20 mL) and extracted with CHCl₂ / CH₃OH (3:1, 3 × 25 mL). The combined organic layers were dried over sodium sulfate and filtered. The solvent was removed under reduced pressure, and the residue was purified by flash column chromatography using a Buchi C-850 (XSelect® CSH™ Prep OBD™ C18 0.5 μM, 50 mm × 150 mm, 100 mL / min, 5–100% CH₃CN in HO (+0.15% TFA) in 30 min) to yield 324 mg (325 μmol, 94%) of an off-white solid. LCMS (Method C) retention time 2.93 min, [M+H] + =882.6. 1H-NMR: (300MHz,DMSO-d6)δ[ppm]=9.83(s,1H), 8.15~8.07(m,1H), 7.91(s,1H), 7.69~7.63(m,1H), 7.45~7.38(m,1H) ), 7.34(s,1H), 7.29~7.23(m,1H), 6.69~6.63(m,1H), 6.49~6.43(m,1H), 5.91~5.67(m,2H), 5.31(d,J=4.9Hz,2H), 4 .91(d,J=4.8Hz,2H), 4.58~4.44(m,4H), 3.98(t,J=5.8Hz,2H), 3.92~3.86(m,5H), 3.68(s,3H), 3.65~3.51(m,2H), 3 .40~3.23(m,2H), 3.16(t,J=8.0Hz,2H), 3.02~2.96(m,2H), 2.16~2.07(m,6H), 1.98~1.86(m,2H), 1.35~1.20(m,6H).

[0550] Preparation of (Z)-3-((E)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-4-methoxy-2,3-dihydrobenzo[d]thiazole-6-carboxylic acid (XX3.8) [ka]

[0551] Compound XX3.7 (100 mg, 100 μmol) was dissolved in CHOH / THF / H0 (2:2:1, 2.5 mL). LiOH (14 mg, 602 μmol) was then added, and the resulting reaction mixture was stirred at room temperature for 18 h. The reaction progress was monitored by LC / MS. The organic solvent was removed under reduced pressure. The aqueous phase was cooled to 0 °C and neutralized with 1 M aqueous HCl. The precipitate was filtered and washed with water. The solid was dissolved / suspended in CHCN / H0 (1:1, 15 mL) and freeze-dried to give 74 mg (85 μmol, 85%) of a white solid. The product was used without further purification. LCMS (Method A) retention time 1.96 min, [M+H] + =868.6. 1 H-NMR: (300MHz,DMSO-d6)δ[ppm]=13.16(brs,1H), 12.84(brs,1H), 10.77(brs,1H), 8.09(d,J=1.5Hz,1H), 7.93(s,1H), 7.6 6(d,J=1.2Hz,1H), 7.50(d,J=1.5Hz,1H), 7.32(d,J=1.4Hz,2H), 6.63(s,1H), 6.46(s,1H), 5.97~5.83(m,1H), 5.84~5.69(m, 1H), 5.31(d,J=5.3Hz,2H), 4.94(d,J=4.9Hz,2H), 4.51(q,J=7.1Hz,4H), 4.04(t,J=5.9Hz,2H), 3.94~3.83(m,2H), 3.79~3.6 6(m,5H), 3.31~3.21(m,2H), 3.21~3.05(m,2H), 3.01~2.91(m,2H), 2.14~2.06(m,5H), 2.05~1.92(m,2H), 1.34~1.20(m,6H).

[0552] Preparation of 1-((E)-4-((Z)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-6-(hydrazinecarbonyl)-4-methoxybenzo[d]thiazol-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide trifluoroacetate (Compound XXXIV) [ka]

[0553] BeSp-0708 / 1 (72.0 mg, 82.9 μmol) was suspended in DMF (3 mL). DIPEA (43.3 μL, 248 μmol), NHS (19.5 mg, 165 μmol), and DIC (25.7 μL, 165 μmol) were then added, and the resulting reaction mixture was stirred at 45 °C for 3 h. The reaction mixture was then stirred at room temperature for 17 h. The reaction progress was monitored by LC / MS. 1 M hydrazine in THF (414 μL, 414 μmol) and DIPEA (72.2 μL, 141 μmol) were then added, and the resulting reaction mixture was stirred at room temperature for 1 h (start: October 6, 2023, 8:00 AM). The reaction progress was monitored by LC / MS. The solvent was removed under reduced pressure. The residue was purified by preparative HPLC (Method I) on RP18 silica. The product-containing fractions were freeze-dried to give 41.0 mg (41.2 μmol, 79%, 96.7% purity by HPLC Method A) of a white solid. LCMS (Method C) retention time 2.94 min, [M+H] + =882.5. [Example]

[0554] Synthesis scheme of compound XXXV [ka]

[0555] Preparation of tert-butyl (E)-(4-((4-carbamoyl-2-(3-morpholinopropoxy)-6-nitrophenyl)amino)but-2-en-1-yl)carbamate (XX1.1) [ka]

[0556] To a suspension of tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (4.27 g, 22.9 mmol) and 4-chloro-3-(3-morpholinopropoxy)-5-nitrobenzamide (5.25 g, 15.3 mmol) in n-butanol (100 mL) was added DIPEA (6.01 mL, 48.9 mmol), and the resulting reaction mixture was heated to 120 °C for 48 h. The solvent was removed in vacuo. The residue was purified on silica by automated flash column chromatography using a Biotage Selekt (Biotage Sfar Silica HC, 20 μM, 100 g, 100 mL / min, 5–15% CHOH in CHCl for 30 min). Product-containing fractions were combined to yield 7.02 g (14.2 mmol, 93%) of a red solid. LCMS (Method C) retention time 2.97 min, [M+H] + =494.1. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=8.19(d,J=1.9Hz,1H), 8.02(brs,1H), 7.76(t,J=6.1Hz,1H), 7.56(d,J=1.9Hz,1H), 7.29(brs,1H), 6 .92(t,J=5.9Hz,1H), 5.65~5.48(m,2H), 4.19~4.05(m,4H), 3.67~3.41(m,6H), 2.48~2.28(m,6H), 2.03~1.90(m,2H), 1.41~1.19(m,9H).

[0557] Preparation of tert-butyl (E)-(4-((2-amino-4-carbamoyl-6-(3-morpholinopropoxy)phenyl)amino)but-2-en-1-yl)carbamate (XX1.2) [ka]

[0558] To a stirred solution of compound XX.1 (7.02 g, 14.2 mmol) in CHOH (210 mL) was added sodium dithionite (24.8 g, 142 mmol) dissolved in water (70 mL) at 0 °C. Then, NH (30% aqueous solution, 14.2 mL, 107 mmol) was added to the mixture at 0 °C. The reaction was allowed to warm to room temperature and stirred at room temperature for 2 h. The progress of the reaction was monitored by LCMS. The reaction was diluted with H0 (500 mL) and CHCl (700 mL) was added. The aqueous layer was extracted with CHCl / CHOH (2 × 300 mL). The organic layer was dried over NaSO and filtered. The solvent was removed under reduced pressure to give 5.00 g (10.8 mmol, 76%) of a yellowish foam. The compound was used without further purification. LCMS (Method C) retention time 1.78 min, [M+H] + =464.1. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=7.58(brs,1H), 7.04~6.81(m,3H), 6.78(d,J=1.9Hz,1H), 5.69~5.47(m,2H), 4.65(brs ,2H), 3.98(t,J=6.2Hz,2H), 3.81(t,J=7.0Hz,1H), 3.63~3.43(m,8H), 2.47~2.30(m,6H), 1.96~1.81(m,2H), 1.37(s,9H).

[0559] Preparation of tert-butyl (E)-(4-(2-amino-5-carbamoyl-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (XX1.3) [ka]

[0560] Compound XX1.2 (5.00 g, 10.8 mmol) was dissolved in CHOH (100 mL) and cooled to 0 °C. Cyanogen bromide (1.94 g, 18.3 mmol) was then added, and the mixture was allowed to warm to room temperature. The resulting mixture was stirred at room temperature for 24 h. The reaction progress was monitored by LCMS. All volatiles were removed under reduced pressure. The residue was purified by automated flash column chromatography on silica using a Biotage Selekt (Biotage Sfar Silica HC, 20 μM, 200 g, 120 mL / min, 5–15% CHOH in CHCl (+10% aqueous NHOH (35%)) for 45 min) to give 3.46 g (7.08 mmol, 66%). LCMS (Method C) retention time 1.86 min, [M+H] + =489.1. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=7.77(brs,1H), 7.39(d,J=1.3Hz,1H), 7.13~6.97(m,2H), 6.96~6.85(m,1H), 6.38(s,2H), 5.71~5.42(m, 2H), 4.78(d,J=5.4Hz,2H), 4.11(t,J=6.3Hz,2H), 3.63~3.54(m,4H), 3.53~3.43(m,2H), 2.49~2.32(m,6H), 2.01~1.86(m,2H), 1.34(s,9H).

[0561] Preparation of tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate trifluoroacetate (XX1.4) [ka]

[0562] Compound XX1.3 (1.50 g, 3.07 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (521 mg, 3.38 mmol) were dissolved in DMF (20 mL) and then treated with DIPEA (2.03 mL, 12.3 mmol) at 0 °C for 15 min. HATU (1.52 g, 3.99 mmol) was then added, and the resulting yellowish reaction mixture was stirred at room temperature for 1 h. Product formation was monitored by LCMS. The solvent was removed in vacuo, and the residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CH3CN in HO (+0.15% TFA) for 47 min). The product-containing fractions were combined and freeze-dried to give 1.91 g (2.59 mmol, 84%) of a colorless solid. LCMS (Method C) retention time 3.15 min, [M+H] + =625.2. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=9.78(brs,1H), 7.97(brs,1H), 7.69(d,J=1.2Hz,1H), 7.44 ~7.32(m,2H), 6.91(brs,1H), 6.66~6.60(m,1H), 5.84~5.69(m,1H), 5.58~5.43(m,1H), 5.03~4 .89(m,2H), 4.68~4.54(m,2H), 4.27(t,J=5.9Hz,2H), 4.11~3.96(m,2H), 3.76~3.60(m,2H), 3. 60~3.42(m,4H), 3.38~3.26(m,2H), 3.25~3.06(m,2H), 2.33~2.11(m,5H), 1.43~1.08(m,12H).

[0563] Preparation of (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide hydrochloride (XX1.5) [ka]

[0564] Compound XX1.4 (1.90 g, 2.57 mmol) was dissolved / suspended in CHCl (5 mL). 4 M HCl in dioxane (5.14 mL) was then added, and the resulting mixture was stirred at room temperature for 2.5 h. The reaction progress was monitored by LCMS. Aqueous HCl (0.15% in HO) was then added, and all volatiles were then removed in vacuo. The residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CHCN in HO (+0.15% HCl) over 47 min). The product-containing fractions were combined and freeze-dried to give 1.41 g (2.51 mmol, 98%) of a white solid. LCMS (Method C) retention time 2.03 min, [M+H] + =525.1. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=11.65(s,1H), 8.26(brs,3H), 7.71(d,J=1.2H z,1H), 7.52~7.29(m,2H), 6.74(s,1H), 6.19~6.05(m,1H), 5.70~5.55(m,1H), 5. 04(d,J=5.4Hz,2H), 4.68~4.54(m,2H), 4.31(t,J=6.1Hz,2H), 4.04~3.84(m,4H) , 3.59~3.06(m,9H), 2.45~2.30(m,2H), 2.23~2.17(m,3H), 1.36(t,J=7.1Hz,3H).

[0565] Preparation of methyl (E)-5-bromo-6-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)nicotinate trifluoroacetate (XX1.6) [ka]

[0566] Compound XX1.5 (1.29 g, 2.30 mmol) and methyl 5-bromo-6-chloronicotinate (748.7 mg, 2.99 mmol) were dissolved in DMF (40 mL). CsCO (3.05 g, 9.20 mmol) was then added, and the resulting reaction mixture was heated to 80 °C for 18 h. Product formation was monitored by LCMS. The suspension was filtered, and the solvent was removed in vacuo. The residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 250 mm, 100 mL / min, 5–100% CH3CN in HO (+0.15% TFA) for 45 min). The product-containing fractions were combined and freeze-dried to yield 648 mg (33%, 759 μmol) of a white solid. LCMS (Method C) Retention time 3.56 min, [M+2H] 2+ =369.5. 1 H-NMR (300MHz, DMSO-d6): d[ppm]=12.76(brs,1H), 9.99(brs,1H), 8.47(d,J=2.0Hz,1H), 8.07(d,J=2.0Hz,1H ), 7.95(brs,1H), 7.68(d,J=1.2Hz,1H), 7.41~7.25(m,3H), 6.56(d,J=0.6Hz,1H), 5.89~5.61(m,2H), 4.94(d,J =5.3Hz,2H), 4.56(q,J=7.1Hz,2H), 4.21(t,J=5.9Hz,2H), 4.08~3.93(m,4H), 3.79(s,3H), 3.67(t,J=12.1Hz,2 H), 3.46(d,J=12.2Hz,2H), 3.29(t,J=8.0Hz,2H), 3.21~3.01(m,2H), 2.23~2.12(m,5H), 1.30(t,J=7.1Hz,3H).

[0567] Preparation of methyl (E)-6-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)-5-((3-methoxy-3-oxopropyl)thio)nicotinate trifluoroacetate (XX1.7) [ka]

[0568] Methyl (E)-5-bromo-6-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)nicotinate (618 mg, 725 μmol) was co-evaporated with anhydrous 1,4-dioxane (5 mL). The residue was dissolved in anhydrous 1,4-dioxane (7.5 mL), and DIPEA (372 μL, 2.17 mmol) and methyl 3-mercaptopropanoate (321 μL, 2.90 mmol) were added. Xantphos (41.9 mg, 72.5 μmol) and Pd(dba) (33.2 mg, 36.2 μmol) were then added under a slight argon flow, and the resulting mixture was heated to 130 °C (hotplate temperature) for 2 h. Product formation was monitored by LCMS. Water was added, and then all solvent was removed in vacuo. The residue (filtered through a syringe filter PVDF-45 / 25) was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 250 mm, 100 mL / min, 10–100% CH3CN in HO (+0.15% TFA) for 47 min). The fractions containing the product were freeze-dried (CHCN / H0 = 1:1) to give 424 mg (475 μmol, 66%) of a white solid. LCMS (Method C) retention time 2.95 min, [M+H] + =778.2. 1H-NMR (300MHz, DMSO-d6): d[ppm]=12.82(s,1H), 9.98(s,1H), 8.49(d,J=2.2Hz,1H), 7.98~7.88(m,2H), 7.67(d,J= 1.2Hz,1H), 7.42~7.21(m,3H), 6.55(d,J=0.6Hz,1H), 5.89~5.61(m,2H), 4.94(d,J=5.2Hz,2H), 4.56(q,J=7.1Hz,2 H), 4.21(t,J=5.9Hz,2H), 4.11~3.91(m,4H), 3.79(s,3H), 3.67(t,J=12.1Hz,2H), 3.53~3.40(m,5H), 3.29(t,J=8. 0Hz,2H), 3.21~3.01(m,2H), 2.87(t,J=6.8Hz,2H), 2.45(t,J=6.8Hz,2H), 2.22~2.11(m,5H), 1.30(t,J=7.1Hz,3H).

[0569] Preparation of sodium (E)-2-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)-5-(methoxycarbonyl)pyridine-3-thiolate (XX1.8) [ka]

[0570] Methyl (E)-6-((4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)amino)-5-((3-methoxy-3-oxopropyl)thio)nicotinate (400 mg, 449 μmol) was dissolved in anhydrous THF (5 mL), 25% NaOCH in CHOH (410 μL, 1.79 mmol) was added under a slight argon flow, and the resulting mixture was stirred for 2 h. Product formation was controlled by LCMS. The mixture was diluted with CHCl (5 mL), and the resulting solid was filtered and washed with CHCl (3 × 2 mL). The solid was suspended in CHCN (5 mL) and stirred at room temperature for 1 h. The solid was filtered and washed with cold CH3CN (2 x 2 mL). After drying under high vacuum, the crude product (311 mg) was used without further purification. LCMS (Method C) retention time 2.22 min, [M+H] + =692.2.

[0571] Preparation of methyl (E)-3-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-imino-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (XX1.9) [ka]

[0572] Compound XX.8 (300 mg, 420 μmol) was dissolved in CHOH (5 mL) and cooled to 0 °C. Cyanogen bromide (53.4 mg, 504 μmol) was then added and the mixture was stirred at 0 °C for 1 h. The mixture was then warmed to room temperature and stirred for 1 h. The progress of the reaction was monitored by LCMS. The resulting solid was filtered and washed with ice-cold CHOH (3 mL). The solid (358 mg) was dried in vacuo and used without further purification. LCMS (Method C) retention time 1.99 min, [M+H] + =717.2.

[0573] Preparation of methyl (E)-3-((E)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate trifluoroacetate (XX1.10) [ka]

[0574] Compound XX.9 (300 mg, 419 mmol) and 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (71.0 mg, 460 μmol) were dissolved in DMF (5 mL) and then treated with DIPEA (277 μL, 1.67 mmol) at 0 °C for 15 min. HATU (207 mg, 544 μmol) was then added, and the resulting yellowish reaction mixture was stirred at room temperature for 1 h. Product formation was monitored by LCMS. The solvent was removed in vacuo, and the residue was purified by automated flash column chromatography on RP18 silica using a Buchi C-850 (XSelect® CSH™ Prep OBD™ 0.5 μM, 50 mm × 150 mm, 100 mL / min, 10–100% CH3CN in HO (+0.15% TFA) for 47 min). The product-containing fractions were combined and freeze-dried (CHCN / HO = 1:1) to give 123 mg (127.0 μmol, 34% over 3 steps) of a white solid. LCMS (Method C) retention time 3.32 min, [M+H] + =853.2.

[0575] Preparation of (E)-3-((E)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylic acid (XX1.11) [ka]

[0576] Methyl (E)-3-((E)-4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-2,3-dihydrothiazolo[4,5-b]pyridine-6-carboxylate (122 mg, 126 μmol) was dissolved in CHOH / THF / HO (2:2:1, 5 mL). LiOH (9.06 mg, 378 μmol) was then added, and the resulting reaction mixture was stirred at room temperature for 4 h. The reaction progress was monitored by LCMS. The organic solvent was removed in vacuo (rotary evaporator). The aqueous phase was cooled to 0°C and neutralized with 1M aqueous HCl. The precipitate was filtered and washed with water (2 mL). The solid was dissolved / suspended in CHCN / H0 (1:1, 25 mL) and freeze-dried. The resulting product (90.8 mg) was used without further purification. LCMS (Method C) retention time 3.13 min, [M+2H] 2+ =420.2. 1H-NMR (300MHz, DMSO-d6): d[ppm]=12.80(s,1H), 8.93(d,J=1.9Hz,1H), 8.85(d,J=1.9Hz,1H), 7.92(s,1 H), 7.63(s,1H), 7.34~7.28(m,2H), 6.61(d,J=0.7Hz,1H), 6.45(s,1H), 6.02(dt,J=15.5,5.3Hz,1H), 5.7 9(dd,J=15.2,6.2Hz,1H), 5.07(d,J=5.7Hz,2H), 4.94(d,J=5.2Hz,2H), 4.58~4.42(m,4H), 4.08(t,J=5. 9Hz,2H), 3.73~3.65(m,5H), 3.05~2.63(m,4H), 2.19~2.03(m,6H), 1.96~1.90(m,2H), 1.33~1.17(m,6H).

[0577] Preparation of 1-((E)-4-((E)-2-((1-ethyl-3-methyl-1H-pyrazole-5-carbonyl)imino)-6-(hydrazinecarbonyl)thiazolo[4,5-b]pyridin-3(2H)-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-7-(3-morpholinopropoxy)-1H-benzo[d]imidazole-5-carboxamide trifluoroacetate (Compound XXXV) [ka]

[0578] Compound XX.11 (51.7 mg) was suspended in DMF (3 mL). Then, DIPEA (32.2 μL), NHS (14.5 mg), and DIC (19.1 μL) were added, and the resulting reaction mixture was stirred at 40 °C for 3 h. The reaction was then stirred at room temperature for 15 h. The reaction progress was monitored by LC / MS. Next, 1 M hydrazine in THF (308 μL) and DIPEA (32.2 μL) were added, and the resulting reaction mixture was stirred at room temperature for 1 h. The reaction progress was monitored by LC / MS. The solvent was removed in vacuo (oil pump vacuum) and freeze-dried (CHCN / HO 1:1, 10 mL). The residue was purified by preparative HPLC (Method I) on RP18 silica. The product-containing fractions were freeze-dried to give 46.9 mg (79%, 95.9% purity by HPLC Method A) of a white solid. [Example]

[0579] PBMC and THP-1 assays of compounds XXVIII-XXXV The PBMC assay was performed according to the method described above in Example 23. The activity of compounds XXVIII-XXXV is shown in FIG.

[0580] A THP-1 assay was further performed to assess the membrane permeability of the compounds according to the method described above in Example 20. The results are shown in Figure 14 below. [Example]

[0581] Synthesis of Compound XL [ka]

[0582] Preparation of methyl 4-{[(2E)-4-{[(tert-butoxy)carbonyl]amino}but-2-en-1-yl]amino}-3-methoxy-5-nitrobenzoate (XX1-2) [ka]

[0583] To a solution of methyl 4-chloro-3-methoxy-5-nitrobenzoate XX1-1 (1.00 g, 4.07 mmol, 1.00 equiv.) and tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (834 mg, 4.48 mmol, 1.10 equiv.) in tetrahydrofuran (15 mL), potassium carbonate (1.13 g, 8.14 mmol, 2.00 equiv.) was added at room temperature. The reaction mixture was stirred at 60 °C for 64 hours. The resulting mixture was concentrated in vacuo. The residue was redissolved in ethyl acetate (100 mL) and washed with water (2 × 50 mL) and saturated aqueous sodium chloride solution (100 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated in vacuo to give pure compound XX1-2 (1.61 g, 4.07 mmol, quantitative) as an orange solid. 1 H NMR (DMSO-d6,400MHz): 1.34(s,9H), 3.42~3.50(m,2H), 3.83(s,3H), 3.89(s,3H), 4.11~4.13(m,2H), 5.52~5.53(m,2H), 6.56(br s,0.1H), 6.92(t,0.9H,J=5.4Hz), 7.43(d,1H,J=1.8Hz), 8.01(t,1H,J=6.1Hz), 8.16(t,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%FA): Retention time=3.23 min; [M+Na + ]=418.4.

[0584] Preparation of methyl 4-{[(2E)-4-aminobut-2-en-1-yl]amino}-3-methoxy-5-nitrobenzoate hydrochloride (XX1-3) [ka]

[0585] To a suspension of compound XX1-2 (3.25 g, 7.23 mmol, 1.00 equiv.) in MeOH (7.15 mL), HCl (4N in dioxane) (14.3 mL, 57.2 mmol, 7.90 equiv.) was added dropwise at room temperature. The reaction mixture was stirred at room temperature for 1.5 hours. The precipitate was filtered and washed with diethyl ether (3 × 20 mL). The filtrate was filtered again and washed with diethyl ether (10 mL). The solids were combined and dried under reduced pressure to give pure crude compound XX1-3 (2.38 g, 7.11 mmol, 98%) as an orange solid. 1 H NMR (DMSO-d6,400MHz): 3.38~3.42(m,2H), 3.84(s,3H), 3.90(s,3H), 4.21(t,2H) ,J=5.8Hz), 5.59~5.66(m,1H), 5.83~5.90(m,1H), 7.46(d,1H,J=1.8Hz), 7.99(br s,3H), 8.13(t,1H,J=6.4Hz), 8.19(d,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%FA): Retention time=2.43 min; [M-HCl+H + ]=296.4.

[0586] Preparation of 4-chloro-3-methoxy-5-nitrobenzamide (XX1-4) [ka]

[0587] A suspension of methyl 4-chloro-3-methoxy-5-nitrobenzoate XX1-1 (50.0 g, 204 mmol, 1.00 equiv) in 30% aqueous ammonium hydroxide solution (733 mL) was stirred at 50° C. for 1.5 hours (sealed vessel) and then at room temperature for 16 hours. The mixture was then stirred at 50° C. for 24 hours (open vessel). The reaction mixture was cooled to room temperature and filtered. The solid was washed with water (500 mL), diethyl ether (400 mL), and dried (50° C., 61 hours) to give pure compound XX1-4 (40.6 g, 176 mmol, 86%) as a yellow solid. 1H NMR (DMSO-d6,400MHz): δ4.02(s,3H), 7.78(brs,1H), 7.88(d,1H,J=1.7Hz), 8.05(d,1H,J=1.7Hz), 8.29(brs,1H). LCMS (2~100 ACN / H2O+0.1%FA): Retention time=2.66 min; [M+H + ]=231.2.

[0588] Preparation of 4-chloro-3-hydroxy-5-nitrobenzamide (XX1-5) [ka] To a solution of compound XX1-4 (31.5 g, 136 mmol, 1.00 equiv.) in dichloromethane (400 mL) cooled to 0 °C, boron tribromide (1 M in DCM, 545 mL, 545 mmol, 4.00 equiv.) was added over 20 minutes, and the mixture was stirred at 40 °C for 42 hours. The reaction mixture was allowed to reach room temperature, poured into 500 mL of ice / water, and stirred for 10 minutes (until the ice melted). The precipitate was filtered, washed twice with water and twice with n-pentane, and dried under reduced pressure (50 °C, 16 hours) to give a white solid (41.4 g). The residue was triturated and ground in water (400 mL) for 1 hour and filtered. The filter cake was washed with water (500 mL) and n-pentane (2 × 500 mL) and dried under vacuum to give a white solid (25.3 g). The residue was triturated and triturated in water (400 mL) for 2 hours, then filtered. The filter cake was washed with water (500 mL) and n-pentane (2 × 500 mL) and dried under vacuum to give crude compound XX1-5 (23.5 g, 109 mmol, 80%) as a white solid. 1 H NMR (DMSO-d6,400MHz): δ7.67(brs,1H), 7.72(d,1H,J=1.8Hz), 7.93(d,1H,J=1.9Hz), 8.18(brs,1H), 11.53(brs,1H). LCMS (2~100 ACN+0.1%AF / H2O+0.1%AF): Retention time=2.49min; [M+H + ]=217.2.

[0589] Preparation of 4-chloro-3-[3-(morpholin-4-yl)propoxy]-5-nitrobenzamide (XX1-6) [ka]

[0590] A mixture of compound XX1-5 (1.00 g, 4.62 mmol, 1.00 equiv.), 4-(3-chloropropyl)morpholine (907 mg, 5.54 mmol, 1.20 equiv.), and potassium carbonate (830 mg, 6.00 mmol, 1.30 equiv.) in N,N-dimethylformamide (6.00 mL) was stirred at 70 °C for 16 h. The solvent was removed in vacuo to give a yellow solid (2.71 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 80 g) using dichloromethane / methanol (100:0 to 85:15 over 30 min, then 85:15 for 10 min) to give pure compound XX1-6 (1.17 g, 3.40 mmol, 74%) as a pale yellow solid. 1 H NMR (DMSO-d6,400MHz): δ1.92~1.98(m,2H), 2.35~2.37(m,4H), 2.45(t,2H,J=7.1Hz), 3.56(t,4H,J=4.6Hz), 4.28(t,2H,J=6.3Hz), 7.77(br s,1H), 7.87(d,1H,J=1.7Hz), 8.04(d,1H,J=1.7Hz), 8.27(br s,1H). LCMS (2~100 ACN / H2O+0.1%AF): Retention time=2.37 min; [M+H + ]=344.3.

[0591] Preparation of 4-{[(2E)-4-({4-carbamoyl-2-[3-(morpholin-4-yl)propoxy]-6-nitrophenyl}amino)but-2-en-1-yl]amino}-3-methoxy-5-nitrobenzamide (XX1-7) [ka]

[0592] To a stirred solution of compound XX1-3 (1.06 g, 3.20 mmol, 1.10 equiv.) and compound XX1-6 (1.00 g, 2.91 mmol, 1.00 equiv.) in tetrahydrofuran (10.7 mL), potassium carbonate (804 mg, 5.82 mmol, 2.00 equiv.) and DIPEA (1.68 mL, 10.2 mmol, 3.49 equiv.) were added. The reaction mixture was stirred in a sealed vessel at 100 °C for 3 days. The reaction mixture was cooled to room temperature, diluted with water (40 mL), and extracted with a mixture of dichloromethane / methanol (8 / 2, 3 × 40 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to give an orange solid (2.17 g). The residue was purified by flash chromatography on silica gel (50 μm amorphous, 80 g) using dichloromethane / methanol (98:2 for 10 min, 92:8 for 20 min, then 92:8 for 10 min) to give pure compound XX1-7 (1.17 g, 1.95 mmol, 67%) as an orange solid. 1 H NMR (DMSO-d6,400MHz): δ1.85~1.91(m,2H), 2.32~2.34(m,4H), 2.38(t,2H,J=7.1Hz), 3.55(t,4H,J=4 .5Hz), 3.80(s,3H), 3.84(s,3H), 4.00(t,2H,J=6.3Hz), 4.07~4.11(m,4H), 5.52~5.62(m,2H), 7.30(br s,1H), 7.35(d,1H,J=1.8Hz), 7.47(d,1H,J=1.8Hz), 7.73(t,1H,J=6.2Hz), 7.95~7.98(m,2H), 8.10(d,1H,J=1.8Hz), 8.14(d,1H,J=1.8Hz). LCMS (2~100 ACN / H2O+0.1%AF): Retention time=2.59 min; [M+H + ]=603.5.

[0593] Preparation of methyl 3-amino-4-{[(2E)-4-({2-amino-4-carbamoyl-6-[3-(morpholin-4-yl)propoxy]phenyl}amino)but-2-en-1-yl]amino}-5-methoxybenzoate (XX1-8) [ka]

[0594] To a stirred solution of compound XX1-7 (11.7 g, 19.5 mmol, 1.00 equiv) in MeOH (305 mL) was added sodium dithionite (33.9 g, 195 mmol, 10.0 equiv) dissolved in water (105 mL) at 0 °C. To this stirre...

Claims

1. Compounds of Formula I: 【Chemistry 1】 (I) or a solvate, pharmaceutically acceptable salt, or tautomer thereof [In the formula, Ring A is 【Chemistry 2】 (a) (b) (c) (d) is selected from the group consisting of During the ceremony, G and G １ are independently N, CH, or C—X １ -R ２ and G' and G ２ are independently N or CH; X is N—R, O, or S; X' is N or CH; X １ is CH ２ , O or S; R is hydrogen or C １－４ is alkyl, G and G １ are C-X respectively １ -R ２ If R ２ The groups are linked to form L １ and L and L １ are each independently C ２－４ Alkylene or C ２－４ is alkenylene, R ２ teeth, Hydrogen, C ２－４ Cyclic ethers, C ３－４ Cyclic alkyl, C １－４ Alkyl, 【Transformation 3】 is selected from the group consisting of R １ and R ３ is, independently, 【Chemistry 4】 is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R ８ is —OH or —NR ９ R １０ and R ９ and R １０ are independently hydrogen and C １ -C ６ alkyl, X ２ and X ３ are independently NH or S; Y １ and Y ２ is, independently, 【Transformation 5】 and Z １ , Z ２ , Z ３ , and Z ４ are each independently C, N, O, or S; R ５ , R ６ , and R ７ are independently hydrogen, C １ -C ６ Alkyl, and C ２ -C ６ alkenyl, 【Transformation 6】 wherein R ５ and R ６ may be linked to form a 5- or 6-membered heterocyclic ring; R １５ is —OH or —NR ９ R １０ and Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R １１ , R １２ , R １３ , and R １４ are independently absent, hydrogen, or C １－４ is alkyl, n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, G and G １ At least one of them is C-X １ -R ２ and R ２ teeth, 【Transformation 7】 and / or R １ and R ３ At least one of 【Transformation 8】 is.

2. L is C ２ The compound of claim 1, wherein the alkyl group is -alkenylene.

3. L is C ２ -alkenylene, X ２ and X ３ The compound of claim 1 , wherein is NH. 【Request Item 4】 【Chemistry 9】 【change】 2. The compound of claim 1 selected from the group consisting of: 【Request Item 5】 【Chemistry 10】 【change】 2. The compound of claim 1 selected from the group consisting of:

6. Compound of Formula IA: 【Chemistry 11】 (IA) or a solvate, pharmaceutically acceptable salt, or tautomer thereof [In the formula, X １ is CH ２ , O or S; G is CH, C-SCH ３ , C-OCH ３ , or N, R ２ teeth, Hydrogen, C ２－４ Cyclic ethers, C ３－４ Cyclic alkyl, C １－４ Alkyl, 【Chemistry 12】 is selected from the group consisting of R １ and R ３ is, independently, 【Chemistry 13】 is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R ８ is —OH or —NH ２ and Y １ and Y ２ is, independently, 【Chemistry 14】 and X ５ is S, O, or NR ７ and R ５ , R ６ , and R ７ are independently hydrogen, C １ -C ６ Alkyl, and C ２ -C ６ alkenyl, 【Chemistry 15】 wherein R ５ and R ６ may be linked to form a 5- or 6-membered heterocyclic ring; R １５ is —OH or —NR ９ R １０ and R ９ and R １０ are independently hydrogen and C １ -C ６ alkyl, Ring C is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, R ２ teeth, 【Chemistry 16】 and / or R １ and R ３ At least one of 【Chemistry 17】 is.

7. X １ The compound of claim 6 , wherein is O. 【Request Item 8】 【Chemistry 18】 【change】 7. The compound of claim 6 selected from the group consisting of: 【Request Item 9】 【Chemistry 19】 【change】 7. The compound of claim 6 selected from the group consisting of:

10. Compound of formula IA': 【Chemistry 20】 (IA') or a solvate, pharmaceutically acceptable salt, or tautomer thereof [In the formula, X １ is CH ２ , O or S; G is CH, C-SCH ３ , C-OCH ３ , or N, R ２ teeth, Hydrogen, C ２－４ Cyclic ethers, C ３－４ Cyclic alkyl, C １－４ Alkyl, 【Chemistry 21】 is selected from the group consisting of R １ and R ３ is, independently, 【Chemistry 22】 is selected from the group consisting of Ring B is a 6-membered aromatic ring or a 5- or 6-membered aromatic heterocycle containing 1 to 2 heteroatoms selected from N, O, and S; R ８ is —OH or —NH ２ and Y １ and Y ２ is, independently, 【Chemistry 23】 and X ５ is S, O, or NR ７ and R ５ , R ６ , and R ７ are independently hydrogen, —OH, C １ -C ６ Alkyl, 【Chemistry 24】 wherein R ５ and R ６ may be linked to form a 5- or 6-membered heterocyclic ring; n, p, q, t, and v are independently integers from 2 to 6; k, l, m, o, u, and w are independently integers from 1 to 6; However, R ２ teeth, 【Chemistry 25】 and / or R １ and R ３ At least one of 【Chemistry 26】 is. 【Request Item 11】 【Chemistry 27】 【change】 11. The compound of claim 10 selected from the group consisting of: 【Request Item 12】 【Chemistry 28】 11. The compound of claim 10 selected from the group consisting of:

13. Compound of formula IB: 【Chemistry 29】 (IB) or a solvate, pharmaceutically acceptable salt, or tautomer thereof [In the formula, R １ and R ３ is, independently, 【Transformation 30】 wherein R ５ and R ６ may be linked to form a 5- or 6-membered ring, R １４ is hydrogen or C １－４ is alkyl, R ５ , R ６ , and R ７ are independently hydrogen, —OH, C １ -C ６ Alkyl, 【Chemistry 31】 is selected from w is an integer from 1 to 6, provided that R １ and R ３ At least one of 【Chemistry 32】 is. 【Request Item 14】 【Chemistry 33】 14. The compound of claim 13 selected from the group consisting of: 【Request Item 15】 【Chemistry 34】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 16】 【Chemistry 35】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 17】 【Chemistry 36】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 18】 【Chemistry 37】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 19】 【Chemistry 38】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 20】 【Chemistry 39】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 21】 【Chemistry 40】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 22】 【Chemistry 41】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 23】 【Chemistry 42】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 24】 【Chemistry 43】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 25】 【Chemistry 44】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 26】 【Chemistry 45】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 27】 【Chemistry 46】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 28】 【Chemistry 47】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 29】 【Chemistry 48】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 30] [Chemistry 49] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 31】 【Chemistry 50】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 32】 【Chemistry 51】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 33】 【Chemistry 52】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 34】 【Chemistry 53】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 35】 【Chemistry 54】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 36】 【Chemistry 55】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 37】 【Transformation 56】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 38】 【Chemistry 57】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 39】 【Chemistry 58】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 40] [Chemistry 59] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 41】 【Transformation 60】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 42】 【Chemistry 61】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 43】 【Chemistry 62】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 44】 【Chemistry 63】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 45】 【Chemistry 64】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 46】 【Chemistry 65】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 47】 【Transformation 66】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 48] [Chemistry 67] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 49] [Transformation 68] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 50] [Transformation 69] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 51】 【Chemistry 70】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 52】 【Chemistry 71】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 53】 【Chemistry 72】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 54】 【Chemistry 73】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 55】 【Chemistry 74】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 56】 【Chemistry 75】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 57】 【Chemistry 76】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 58】 【Chemistry 77】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 59】 【Transformation 78】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. [Request Item 60] [Chemistry 79] or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 61】 【Chemistry 80】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 62】 【Chemistry 81】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 63】 【Chemistry 82】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof.

64. R ５ , R ６ , and R ７ 2. The compound of claim 1, wherein at least one of is hydrogen.

65. R １ and R ３ Both, independently, 【Chemistry 83】 2. The compound of claim 1, wherein:

66. R １ and R ３ Both, independently, 【Chemical 84】 7. The compound of claim 6, wherein:

67. R １ and R ３ Both, independently, 【Chemical 85】 14. The compound of claim 13, wherein:

68. R １ and R ３ Both, independently, 【Chemical 86】 2. The compound of claim 1, wherein:

69. R １ and R ３ Both, independently, 【Transformation 87】 7. The compound of claim 6, wherein:

70. R １ and R ３ Both, independently, 【Chemical 88】 14. The compound of claim 13, wherein:

71. A method for treating a disease, disorder, or condition treatable by activation of STING or mediated by STING protein activation, the method comprising administering a therapeutically effective amount of one or more compounds described in any one of claims 1 to 70.

72. 72. The method of claim 71, wherein the disease, disorder, or condition is cancer.

73. G is N and X １ is O and R １ and R ３ are respectively 【Chemistry 89】 and R ２ but 【Chemistry 90】 wherein n is 2, 3, or 4; and R ５ , R ６ and R ７ are independently H, C １ -C ６ Alkyl and 【Chemistry 91】 is selected from Y １ and Y ２ are respectively 【Chemistry 92】 [In the formula, X ５ is O. 【Request Item 74】 【Chemistry 93】 or a solvate, pharmaceutically acceptable salt, or tautomer thereof. 【Request Item 75】 【Chemistry 94】 74. The compound of claim 73, selected from the group consisting of:

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