Heterocyclic compounds as STING modulators

Abstract

Disclosed herein are heterocyclic compounds that can be used as STING modulators, methods of synthesis, and the use of such compounds in treating a variety of diseases, including cancer.

Description

Technical Field

[0001] The present disclosure relates to heterocyclic compounds and their derivatives that are useful as STING modulators. The present disclosure also relates to methods of synthesis and the use of such compounds in treating a variety of diseases, including cancer. Background Art

[0002] The innate immune system is specialized and acts rapidly in response to different danger signals. It provides the first line of defense against bacteria, viruses, parasites, and other infectious threats by producing soluble bioactive molecules such as cytokines or chemokines. By responding to these damage-associated molecular patterns (DAMPs) or pathogen-associated molecular patterns (PAMPs) mentioned above, the innate immune system is able to provide protection against a wide range of threats to the host.

[0003] It has recently been demonstrated that the major sensor of cytosolic DNA is cyclic GMP-AMP synthase (cGAS). cGAS catalyzes the generation of the cyclic dinucleotide 2'-3'-cGAMP, an atypical second messenger that binds tightly to the ER transmembrane adaptor protein STING (stimulator of interferon genes). cGAMP-bound STING undergoes conformational changes, translocates to the perinuclear compartment, and induces activation of the key transcription factors IRF-3 and NF-kB. This leads to a strong induction of type I interferons and the production of proinflammatory cytokines such as IL-6, TNF-α, and IFN-γ.

[0004] Type I interferons and proinflammatory cytokines can strongly enhance T cell activation by increasing the ability of dendritic cells and macrophages to uptake, process, present, and cross-present antigens to T cells. Compounds that can induce the production of type I interferons can be used as adjuvants in vaccines to enhance specific immune responses to antigens. Compounds that can induce the production of interferons have potential uses in the treatment of human cancers.

[0005] In contrast, it was found that excessive amounts of type I interferon were produced in patients with various forms of autoimmune diseases. Human genetic evidence and support from animal model studies confirm the hypothesis that inhibition of STING leads to a reduction in the amount of type I interferon (which is the cause of autoimmune diseases) (Crow YJ et al., Nat, Genet. [Natural Genetics] 2006, 38, 917-920; Stetson DB et al., Cell [Cell] 2008, 134, 587-598). Therefore, inhibitors of STING provide treatment for patients with chronic type I interferon and proinflammatory cytokine production associated with infection or complex autoimmune diseases. Compounds that bind to STING can act as antagonists and can be used to treat, for example, autoimmune diseases.

[0006] It is envisioned that targeting STING with activators or inhibitors may be a promising approach for treating diseases and conditions in which modulation of the type I IFN pathway is beneficial, including inflammatory, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, and as immunogenic compositions or vaccine adjuvants.

[0007] The compounds of the invention modulate the activity of STING and thus may provide therapeutic benefits in the treatment of diseases, disorders and / or conditions in which modulation of STING is beneficial, such as for inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, and as vaccine adjuvants. Summary of the invention

[0008] One object of the present invention is to provide compounds and derivatives thereof useful as STING modulators.

[0009] Aspect 1: A compound having formula (I),

[0010]

[0011] or a pharmaceutically acceptable salt, stereoisomer, tautomer or prodrug thereof,

[0012] in:

[0013] R 1 , R 2 , R 3 , R 4 , R 6 , R 7 and R 8 may be the same or different, and are independently selected from hydrogen, halogen, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CN, -NO2, -OR a 、-SO2R a 、-C(=O)R a 、-C(=O)OR a 、-C(=O)NR a R b 、-C(=NR a )NR b R c 、-NR a R b 、-NR a C(=O)R b 、-NR a C(=O)NR b R c 、-NR aC(=O)OR b 、-NR a SONR b R c 、-NR a SO2NR b R c 、-SO2NR a R b , or -NR a S02R b , the -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Each of the alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl groups is optionally substituted by R d replace;

[0014] R 5 Selected from -R 5a 、-OR 5a 、-SR 5a or -NR 5a R 5b ;

[0015] R 5a and R 5b are independently selected from hydrogen, -C 1-8 Alkyl, -C(=O)R, -(CH2) n1 -NRR', -(CH2CH2O) n1 -NRR', -(OCH2CH2) n1 -NRR', -(CH2OCH2) n1 -NRR' or

[0016] R and R' are each independently selected from hydrogen, -C 1-8 Alkyl, -C(=O)OR 9 、-C(=O)-CH2-OR 9 、-(CH2)2-OR 9 ;

[0017] R 9 Selected from hydrogen, -C 1-8 Alkyl (e.g. tert-butyl), or -NR 9a R 9b ;

[0018] R 9a and R 9b are each independently selected from hydrogen, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;

[0019] Xa and Xb are independently selected from N and CH;

[0020] n1 is 1, 2, 3, 4, 5, or 6;

[0021] m1 and m2 are each independently 0, 1, or 2;

[0022] R a , R b and R c are independently selected from the group consisting of: hydrogen, -C 1-8 Alkyl, -C(=O)OR e or -C(=O)R e , where -C 1-8 The alkyl group is optionally replaced by NR a R b , -C 3-8 cycloalkyl or a 4- to 7-membered heterocyclic ring containing at least one heteroatom selected from N, O and S, wherein the remaining ring atoms are carbon; and

[0023] R d and R e are each independently selected from halogen, -C 1-8 Alkyl, -OH, -NH2, -CN, -C 1-8 Alkoxy, and -C 3-8 Cycloalkyl, where -C 1-8 Alkyl, -C 1-8 Alkoxy and -C 3-8 Each of the cycloalkyl groups is optionally substituted with halogen, -OH, -NH2, -CN, or an oxo group.

[0024] Aspect 2: The compound according to Aspect 1, wherein R 5 Selected from -O-(CH2) n1 -NRR', -(OCH2CH2) n1 -NRR' or -O-(CH2OCH2) n1 -NRR'.

[0025] Aspect 3: The compound according to Aspect 2, wherein R 1 , R 2 , R 3 , R 6 , R 7 and R 8 are each independently selected from the group consisting of: hydrogen or -C 1-8 alkyl.

[0026] Aspect 4: A compound according to any one of aspects 1 to 3, wherein R 4 It's hydrogen.

[0027] Aspect 5: The compound according to aspects 1-4, wherein R 3 and R 8 It's hydrogen.

[0028] Aspect 6: A compound according to any one of aspects 1 to 5, wherein R 1 , R 2 , R 6 and R 7 Each is independently selected from methyl or ethyl.

[0029] Aspect 7: A compound according to any one of aspects 1 to 6, wherein R 1 and R 6 are each ethyl; and / or R 2 and R 7 Each is a methyl group.

[0030] Aspect 8: A compound according to any one of aspects 1 to 7, wherein R 5 Selected from -O-(CH2) n1 -NHR, or -(OCH2CH2) n1 -NHR; n1 is 2, 3 or 4; R is selected from hydrogen, CH3, -C(=O)OR 9 , or -C(=O)-CH2-OR 9 ; R 9 Selected from hydrogen, NH2, or -C(CH3)3.

[0031] Aspect 9: The compound according to aspect 8, wherein R is selected from hydrogen, -CH3, Or -C(=O)OC(CH3)3.

[0032] Aspect 10: A compound according to any one of aspects 1 to 9, wherein formula (I) is

[0033]

[0034] Aspect 11: The compound according to Aspect 10, wherein formula (II) is

[0035]

[0036] Wherein R is selected from CH3,

[0037] Aspect 12: The compound according to aspect 1, wherein the compound is

[0038]

[0039]

[0040]

[0041]

[0042] or a pharmaceutically acceptable salt, stereoisomer, tautomer or prodrug thereof.

[0043] In a second aspect, disclosed herein is a pharmaceutical composition comprising a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0044] In a third aspect, disclosed herein is a method for treating a disease that can be affected by STING modulation, the method comprising administering to a subject in need thereof an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt, stereoisomer, tautomer or prodrug thereof.

[0045] The disease is selected from inflammatory, allergic and autoimmune diseases, infectious diseases, cancer, or precancerous syndromes.

[0046] In a fourth aspect, disclosed herein is the use of a compound herein or a pharmaceutically acceptable salt, stereoisomer, tautomer or prodrug thereof in the preparation of a medicament for treating a disease that can be affected by STING modulation.

[0047] The disease is an inflammatory, allergic or autoimmune disease, an infectious disease, a cancer, or a precancerous syndrome. DETAILED DESCRIPTION

[0048] The following terms have the indicated meanings throughout this specification:

[0049] Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0050] The following terms have the indicated meanings throughout this specification:

[0051] As used herein, including the appended claims, singular forms of words such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

[0052] Unless the context clearly indicates otherwise, the term "or" means and is used interchangeably with the term "and / or".

[0053] The term "alkyl" includes hydrocarbon groups selected from linear and branched saturated hydrocarbon groups, the hydrocarbon group containing from 1 to 18, such as from 1 to 12, further such as from 1 to 10, further such as from 1 to 8, or from 1 to 6, or from 1 to 4 carbon atoms. 1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr"), 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), 1,1-dimethylethyl or tert-butyl ("t-Bu"), 1-pentyl, 2- pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl groups.

[0054] The term "propyl" includes 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr").

[0055] The term "butyl" includes 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), 1,1-dimethylethyl or tert-butyl ("t-Bu").

[0056] The term "pentyl" includes 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl.

[0057] The term "hexyl" includes 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl.

[0058] The term "alkylene" refers to a divalent alkyl group formed by removing two hydrogen atoms from an alkane. Alkylene includes, but is not limited to, methylene, ethylene, propylene, and the like.

[0059] The term "halogen" includes fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0060] The term "alkenyl" includes hydrocarbon groups selected from straight and branched hydrocarbon groups containing at least one C=C double bond and from 2 to 18, such as from 2 to 8, further such as from 2 to 6 carbon atoms. 2-6 Examples of alkenyl groups include, but are not limited to, ethenyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hexa-1,3-dienyl groups.

[0061] The term "alkenylene" refers to a divalent alkenyl group formed by removing two hydrogen atoms from an alkene. Alkenylene includes, but is not limited to, vinylene, butenylene, and the like.

[0062] The term "alkynyl" includes hydrocarbon groups selected from straight and branched hydrocarbon groups, the hydrocarbon group containing at least one C≡C triple bond and from 2 to 18, such as from 2 to 8, further such as from 2 to 6 carbon atoms. Alkynyl groups (such as C 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl (propargyl), 1-butynyl, 2-butynyl, and 3-butynyl groups.

[0063] The term "alkynylene" refers to a divalent alkynyl group formed by removing two hydrogen atoms from an alkyne. Alkenylene includes, but is not limited to, ethynylene and the like.

[0064] The term "cycloalkyl" includes hydrocarbon groups selected from saturated cyclic hydrocarbon groups including monocyclic and polycyclic (eg, bicyclic and tricyclic) groups (including fused, bridged or spirocyclic alkyl groups).

[0065] For example, the cycloalkyl group can include from 3 to 12, for example from 3 to 10, further for example 3 to 8, further for example 3 to 6, 3 to 5 or 3 to 4 carbon atoms. Even further for example, the cycloalkyl group can be selected from the monocyclic group including from 3 to 12, for example from 3 to 10, further for example 3 to 8, 3 to 6 carbon atoms. The example of monocyclic cycloalkyl group includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl groups. Especially, saturated monocyclic cycloalkyl groups (for example C 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In preferred embodiments, cycloalkyl groups are monocyclic rings (abbreviated as C 3-6Examples of bicyclic cycloalkyl groups include those having from 7 to 12 ring atoms, having a fused bicyclic arrangement (selected from [4,4], [4,5], [5,5], [5,6] and [6,6] ring systems) or having a bridged bicyclic arrangement (selected from bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and bicyclo[3.2.2]nonane). Additional examples of bicyclic cycloalkyl groups include those having a bicyclic arrangement (selected from [5,6] and [6,6] ring systems).

[0066] The term "spirocycloalkyl" includes cyclic structures containing carbon atoms and formed by at least two rings sharing one atom.

[0067] The term "fused cycloalkyl" includes bicyclic cycloalkyl groups as defined herein which are saturated and formed by two or more rings which share two adjacent atoms.

[0068] The term "bridged cycloalkyl" includes a ring structure containing carbon atoms and formed by two rings that share two atoms that are not adjacent to each other. The term "7- to 10-membered bridged cycloalkyl" includes a ring structure containing 7 to 12 carbon atoms and formed by two rings that share two atoms that are not adjacent to each other.

[0069] Examples of fused cycloalkyl, fused cycloalkenyl or fused cycloalkynyl groups include, but are not limited to, bicyclo[1.1.0]butyl, bicyclo[2.1.0]pentyl, bicyclo[3.1.0]hexyl, bicyclo[4.1.0]heptyl, bicyclo[3.3.0]octyl, bicyclo[4.2.0]octyl, decalin, and benzo 3- to 8-membered cycloalkyl, benzoC 4-6 Cycloalkenyl, 2,3-dihydro-1H-indenyl, 1H-indenyl, 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, etc. A preferred embodiment is an 8- to 9-membered fused ring, which refers to a cyclic structure containing 8 to 9 ring atoms in the above examples.

[0070] The term "aryl", used alone or in combination with other terms, includes groups selected from:

[0071] (a) 5-membered and 6-membered carbocyclic aromatic rings, such as phenyl;

[0072] (b) bicyclic ring systems (e.g., 7- to 12-membered bicyclic ring systems) in which at least one ring is carbocyclic and aromatic, such as naphthyl and indanyl; and,

[0073] (c) tricyclic ring systems (eg 10- to 15-membered tricyclic ring systems) in which at least one ring is carbocyclic and aromatic, such as fluorenyl.

[0074] The terms "aromatic hydrocarbon ring" and "aryl" are used interchangeably throughout the disclosure herein. In some embodiments, the monocyclic or bicyclic aromatic hydrocarbon ring has 5 to 10 ring carbon atoms (i.e., C 5-10 Examples of monocyclic or bicyclic aromatic hydrocarbon rings include, but are not limited to, phenyl, naphthalene-1-yl, naphthalene-2-yl, anthracenyl, phenanthryl, etc. In some embodiments, the aromatic hydrocarbon ring is a naphthalene ring (naphthalene-1-yl or naphthalene-2-yl) or a phenyl ring. In some embodiments, the aromatic hydrocarbon ring is a phenyl ring.

[0075] Specifically, the term "bicyclic fused aryl" embraces bicyclic aryl rings as defined herein. A typical bicyclic fused aryl is naphthalene.

[0076] The term "heteroaryl" includes groups selected from:

[0077] (a) a 5-, 6-, or 7-membered aromatic monocyclic ring containing at least one heteroatom (e.g., from 1 to 4, or in some embodiments from 1 to 3, in some embodiments from 1 to 2 heteroatoms) selected from nitrogen (N), sulfur (S), and oxygen (O), wherein the remaining ring atoms are carbon;

[0078] (b) a 7- to 12-membered bicyclic ring containing at least one heteroatom (e.g., from 1 to 4, or in some embodiments from 1 to 3, or in other embodiments 1 or 2 heteroatoms) selected from N, O and S, wherein the remaining ring atoms are carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring; and

[0079] (c) an 11- to 14-membered tricyclic ring containing at least one heteroatom (e.g., from 1 to 4, or in some embodiments from 1 to 3, or in other embodiments 1 or 2 heteroatoms) selected from N, O and S, wherein the remaining ring atoms are carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in an aromatic ring.

[0080] When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to each other. In some embodiments, the total number of S and O atoms in the heteroaryl group does not exceed 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle does not exceed 1. When the heteroaryl group contains more than one heteroatom ring member, these heteroatoms can be the same or different. The nitrogen atoms in one or more rings of the heteroaryl group can be oxidized to form N-oxides.

[0081] Specifically, the term "bicyclic fused heteroaryl" includes 7- to 12-membered, preferably 7- to 10-membered, more preferably 9- or 10-membered fused bicyclic heteroaryl rings as defined herein. Typically, the bicyclic fused heteroaryl is a 5-membered / 5-membered, 5-membered / 6-membered, 6-membered / 6-membered, or 6-membered / 7-membered bicyclic ring. The group may be attached to the rest of the molecule via either ring.

[0082] "Heterocyclyl," "heterocycle," or "heterocyclic" are interchangeable and include non-aromatic heterocyclyl groups containing one or more heteroatoms selected from nitrogen, oxygen, or optionally oxidized sulfur as ring members, wherein the remaining ring members are carbon, including monocyclic, fused, bridged, and spirocyclic rings, i.e., containing monocyclic heterocyclyls, bridged heterocyclyls, spiro heterocyclyls, and fused heterocyclic groups.

[0083] The term "at least one substituent" disclosed herein includes, for example, from 1 to 4, for example, from 1 to 3, further such as 1 or 2 substituents, provided that the valence theory is satisfied. For example, "at least one substituent F" disclosed herein includes from 1 to 4, for example, from 1 to 3, further such as 1 or 2 substituents F.

[0084] The term "divalent" refers to a linking group that can form a covalent bond with two other moieties. For example, a "divalent cycloalkyl group" refers to a cycloalkyl group obtained by removing two hydrogens from the corresponding cycloalkane to form a linking group. The terms "divalent aryl group", "divalent heterocyclyl group" or "divalent heteroaryl group" should be understood in a similar manner.

[0085] The compounds disclosed herein may contain asymmetric centers and therefore may exist as enantiomers. "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of each other. When the compounds disclosed herein have two or more chiral centers, they may additionally exist as diastereomers. Enantiomers and diastereomers belong to a wider class of stereoisomers. It is intended to include all possible stereoisomers, such as substantially pure resolved enantiomers, their racemic mixtures, and mixtures of diastereomers. It is intended to include stereoisomers of all compounds disclosed herein and / or their pharmaceutically acceptable salts. Unless otherwise specifically stated, a reference to an isomer is applicable to any possible isomer. Whenever the composition of an isomer is not specified, all possible isomers are included.

[0086] When compounds disclosed herein contain olefinic double bonds, unless specified otherwise, such double bonds are intended to include both E and Z geometric isomers.

[0087] When the compounds disclosed herein contain a disubstituted ring system, the substituents found on such ring systems can be in cis and trans forms. The cis form means that both substituents are located on the upper side of the 2 substituent positions on the carbon, while the trans form means that they are located on the opposite side. For example, the disubstituted ring system can be a cyclohexyl ring or a cyclobutyl ring.

[0088] It may be advantageous to separate the reaction products from each other and / or from the starting materials. The desired products of each step or series of steps are separated and / or purified (hereinafter referred to as separation) to the desired uniformity by common techniques in the art. Typically, such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can involve many methods, including, for example: reverse phase and normal phase; size exclusion; ion exchange; high, medium, and low pressure liquid chromatography methods and apparatus; small-scale analysis; simulated moving bed ("SMB") and preparative thin or thick layer chromatography, as well as small-scale thin layer and flash chromatography techniques. Those skilled in the art can select and apply the techniques most likely to achieve the desired separation.

[0089] "Diastereomers" refer to stereoisomers of compounds that have two or more chiral centers but are not mirror images of each other. Diastereoisomer mixtures can be separated into their individual diastereomers by methods well known to those skilled in the art such as chromatography and / or fractional crystallization according to their physicochemical differences. Enantiomers can be separated as follows: by reacting with an appropriate optically active compound (e.g., a chiral auxiliary, such as a chiral alcohol or Mosher's acidchlorid) the enantiomeric mixture is converted into a diastereomeric mixture, separating these diastereomers, and converting (e.g., hydrolyzing) a single diastereomer into the corresponding pure enantiomer. Enantiomers can also be separated using a chiral HPLC column.

[0090] Single stereoisomers (e.g., substantially pure enantiomers) can be obtained by resolution of racemic mixtures using an optically active resolving agent to form diastereomers (Eliel, E. and Wilen, S. Stereochemistry of Organic Compounds. New York: John Wiley & Sons, Inc., 1994; Lochmuller, CH et al. "Chromatographic resolution of enantiomers: Selective review." J. Chromatogr., 113(3) (1975): 283-302). Racemic mixtures of chiral compounds of the invention may be separated and isolated by any suitable method, including: (1) forming ionic diastereomeric salts with chiral compounds and separating by fractional crystallization or other methods; (2) forming diastereomeric compounds with chiral derivatizing agents, separating these diastereomers and converting them to pure stereoisomers; and (3) separating substantially pure or enriched stereoisomers directly under chiral conditions. See: Wainer, Irving W., ed., Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.

[0091] Some of the compounds disclosed herein may exist with different points of attachment of hydrogen, known as tautomers. For example, a compound comprising a carbonyl -CH2C(O)- group (keto form) may undergo tautomerism to form a hydroxyl -CH=C(OH)- group (enol form). Where applicable, individual keto and enol forms as well as mixtures thereof are also intended to be included.

[0092] "Prodrug" refers to a derivative of an active agent that requires a transformation in vivo to release the active agent. In some embodiments, the transformation is an enzymatic transformation. Prodrugs are often (although not necessarily) pharmacologically inactive until converted to an active agent.

[0093] "Pharmaceutically acceptable salts" refers to those salts which are suitable for use in contact with the tissues of humans and lower animals within the scope of sound medical judgment without undue toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately prepared by reacting a free base functional group with a suitable organic acid, or separately prepared by reacting an acidic group with a suitable base. The term also includes salts of stereoisomers (e.g., enantiomers and / or diastereomers), tautomers, and prodrugs of the compounds of the invention.

[0094] In addition, if the compounds disclosed herein are obtained as acid addition salts, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt (e.g., a pharmaceutically acceptable addition salt) can be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid according to conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize a variety of synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable addition salts without undue experimentation.

[0095] The terms "administration" and "treating" as used herein, when applied to an animal, a human, an experimental subject, a cell, a tissue, an organ, or a biological fluid, refer to the contact of an exogenous drug, a therapeutic, a diagnostic agent or a composition with an animal, a human, a subject, a cell, a tissue, an organ, or a biological fluid. The treatment of cells encompasses the contact of an agent with a cell and the contact of an agent with a fluid, wherein the fluid is in contact with the cell. The terms "administration" and "treatment" also refer to the in vitro and ex vivo treatment of a cell, such as by an agent, a diagnostic agent, a binding compound, or another cell. The term "subject" as used herein includes any organism, preferably an animal, more preferably a mammal (e.g., rats, mice, dogs, cats, and rabbits), and most preferably a human.

[0096] The term "effective amount" or "therapeutically effective amount" refers to the amount of an active ingredient (e.g., compound) sufficient to affect the treatment of a disease, disorder, or symptom when administered to a subject to treat a disease, or at least one clinical symptom of a disease or disorder. The term "therapeutically effective amount" may vary with the compound, the disease, disorder, and / or the symptoms of the disease or disorder, the severity of the disease, disorder, and / or the symptoms of the disease or disorder, the age of the subject to be treated, and / or the weight of the subject to be treated. The appropriate amount in any given case will be apparent to those skilled in the art, or may be determined by routine experimentation. In some embodiments, a "therapeutically effective amount" is an amount of at least one compound disclosed herein and / or at least one of its stereoisomers, tautomers, or prodrugs, and / or at least one of its pharmaceutically acceptable salts as defined herein that effectively "treats" a disease or disorder in a subject. In the case of combination therapy, the term "therapeutically effective amount" refers to the total amount of a combination object used to effectively treat a disease, disorder, or condition.

[0097] The term "disease" refers to any illness, ailment, disease, symptom, or indication, and is interchangeable with the terms "disorder" or "condition."

[0098] Throughout this specification and the appended claims, unless the context requires otherwise, the term "comprise" and variations such as "comprises and comprising" are intended to specify the presence of the following features, but do not preclude the presence or addition of one or more other features. When used herein, the term "comprises" may be replaced with the terms "containing" or "including", or sometimes with "having".

[0099] Throughout this specification and the appended claims, the term “C n-m " indicates a range including endpoints, where n and m are integers and indicate the number of carbons. Examples include C 1-8 , C 1-6 wait.

[0100] Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0101] Universal synthesis

[0102] The compounds disclosed herein, including salts thereof, can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes.

[0103] The reaction for preparing the compounds disclosed herein can be carried out in a suitable solvent that can be easily selected by a technician in the field of organic synthesis. Suitable solvents can be substantially non-reactive with starting materials, intermediates or products at a temperature (e.g., a temperature ranging from the boiling temperature of the solvent) for carrying out the reaction. A given reaction can be carried out in a solvent or a mixture of multiple solvents.

[0104] The selection of appropriate protecting groups can be readily determined by one skilled in the art.

[0105] The reaction can be monitored according to any suitable method known in the art (e.g., NMR, UV, HPLC, LC-MS, and TLC). The compounds can be purified by a variety of methods including HPLC and normal phase silica gel chromatography.

[0106] Chiral analytical HPLC was used for retention time analysis of different chiral examples, and the conditions were divided into the following methods according to the column, mobile phase and solvent ratio used.

[0107] Scheme I

[0108]

[0109] For example, a compound having formula (I) can be formed as shown in Scheme I. Compound (ii) can be alkylated with compound (i) to give compound (iii) [ie, formula (I)], wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 As defined herein.

[0110] Solution II

[0111]

[0112] For example, compounds having formula (II) can be formed as shown in Scheme II, wherein R 5As defined herein. Compound (i) can be reduced and then acetylated to give compound (ii); compound (ii) can be reacted with malononitrile and then treated with alkaline methanol to give compound (iii); compound (iii) can be condensed with aldehyde (iv) to give compound (v); compound (v) can be reacted with POCl3 to give compound (vi); compound (vi) can be hydrolyzed to give compound (vii); compound (vii) can be dechlorinated to give compound (viii). Compound (ix) may be substituted with compound (x) to give compound (xi); compound (xi) may be reduced to give compound (xii); compound (xii) may be acylated to give compound (xiii); compound (xiii) may be saponified to give compound (xiv); compound (xiv) may be coupled to give compound (xv); compound (xv) may be deprotected to give compound (xvi); compound (xvi) may be deaminated to give compound (xvii); the hydroxyl group on compound (xvii) may be converted to a leaving group to give compound (xviii); compound (xviii) may be reacted with compound (viii) to provide compound (xiv) [i.e., formula (II)].

[0113] Scheme III

[0114]

[0115] For example, compounds having formula (II) can be formed as shown in Scheme III, wherein R 5 As defined herein. Compound (i) can be coupled with compound (ii) using a transition metal catalyzed reaction to give compound (iii); compound (iii) can be coupled with compound (iv) using a transition metal catalyzed reaction to give compound (v); compound (v) can be intramolecularly arylated by a Cadogan reaction to give compound (viii). Compound (xviii) can be obtained by the method shown in Scheme II and then reacted with compound (viii) to give compound (xiv) [i.e., formula (II)].

[0116] abbreviation

[0117] NMR Nuclear Magnetic Resonance

[0118] DMSO Dimethyl sulfoxide

[0119] LC-MS Liquid Chromatography Mass Spectrometry

[0120] HPLC High Performance Liquid Chromatography

[0121] UV

[0122] PE Petroleum Ether

[0123] EA Ethyl acetate

[0124] Boc tert-Butyloxycarbonyl

[0125] DCM Dichloromethane

[0126] HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-Oxidized hexafluorophosphate

[0127] DIPEA, DIEA N,N-Diisopropylethylamine

[0128] DMF N,N-Dimethylformamide

[0129] Et Ethyl

[0130] Ac Acetyl

[0131] THF Tetrahydrofuran

[0132] Bu Butyl

[0133] MTBE Methyl tert-butyl ether

[0134] dppf 1,1'-ferrocenyldiyl-bis(diphenylphosphine)

[0135] IPA Isopropyl Alcohol

[0136] TFA Trifluoroacetic acid

[0137] Example 1-AA

[0138] Tert-butyl (E)-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)(methyl)carbamate

[0139]

[0140] Step 1: 1-Ethyl-3-methyl-1H-pyrazole-5-isothiocyanate

[0141]

[0142] Under nitrogen, oxalyl chloride (10.5g, 82.5mmol) was added dropwise to a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carboxylic acid (12.7g, 82.5mmol) and N,N-dimethylformamide (0.1mL) in dichloromethane (150mL) and then the mixture was stirred at 0°C for 1h. The solvent was evaporated under vacuum and the crude product was co-evaporated twice with dichloromethane (50mL x 2). The crude material was dissolved in anhydrous acetone (20mL) and then added dropwise to a solution of KSCN (8.0g, 82.5mmol) in acetone (100mL) at 0°C. The mixture was stirred at 0°C for 10min. Additional KSCN (1.6g) was added and the mixture was stirred for 20min. Hexane (100mL) was added to the reaction mixture and the biphasic mixture was concentrated to one third of the volume in vacuum. The process of hexane addition (200 mL) and concentration was repeated twice, then hexane (200 mL) was added and the solid was removed by filtration. The solid was rinsed with hexane (50 mL). The combined filtrate was concentrated and the residue was purified by column chromatography (PE / EtOAc=6 / 1) to give the title compound (10.0 g, 62%).

[0143] Step 2: Methyl (E)-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-nitronicotinate

[0144]

[0145] A solution of methyl 6-chloro-5-nitronicotinate (2.4 g, 11 mmol), tert-butyl (E)-(4-aminobut-2-ene-1-yl)carbamate (2.06 g, 11 mmol) and N,N-diisopropylethylamine (2.84 g, 22 mmol) in ethanol (60 mL) was stirred at 55 ° C for 2 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (150 mL), washed with water (50 mL) and brine (50 mL) in sequence, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography (PE / EtOAc=5 / 1) to give the title compound (3.3 g, 82%). 1 H NMR(400MHz,DMSO-d6)δ9.06(s,1H),8.90(d,J=1.8Hz,1H),8.74(d,J=2.0Hz,1H),6 .96(s,1H),5.72-5.51(m,2H),4.22(s,2H),3.85(s,3H),3.51(s,2H),1.35(s,9H). LC-MS(M+H) + =367.1.

[0146] Step 3: Methyl (E)-5-amino-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)nicotinate

[0147]

[0148] To a solution of methyl (E)-6-((4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)amino)-5-nitronicotinate (3.3 g, 9.0 mmol) in methanol (60 mL) was added aqueous ammonia (25%, 1 mL) and a solution of Na2S2O4 (15.7 g, 90 mmol) in water (60 mL) at 0°C and the mixture was stirred at room temperature for 4 h. The mixture was diluted with water (200 mL) and extracted with ethyl acetate (150 mL x 4). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated to give the title compound (1.8 g, 59%). 1 H NMR(400MHz,DMSO-d6)δ8.02(s,1H),7.15(s,1H),6.97(s,1H),6.49(s,1H),5.6 3-5.57(m,2H),4.99(s,2H),4.02(s,2H),3.74(s,3H),3.53(s,2H),1.36(s,9H). LC-MS(M+H) + =337.2.

[0149] Step 4: Methyl (E)-3-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxylate

[0150]

[0151] Over a period of 20 min, to a solution of methyl (E)-5-amino-6-((4-((tert-butoxycarbonyl)amino)but-2-ene-1-yl)amino)nicotinate (1.8 g, 5.34 mmol) in N,N-dimethylformamide (20 mL) was slowly added dropwise a solution of 1-ethyl-3-methyl-1H-pyrazole-5-isothiocyanate carbonyl (937 mg, 4.81 mmol) in 1,4-dioxane (10 mL). The mixture was stirred at room temperature for 15 min, then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.54 g, 8.01 mmol) and triethylamine (1.35 g, 13.35 mmol) were added. The mixture was stirred at room temperature overnight. The mixture was poured into water and extracted with ethyl acetate (100 mL x 4). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography (DCM / MeOH = 150 / 1 to 80 / 1, containing 0.1% aqueous ammonia) to give the title compound (940 mg, 35%). 1 H NMR(400MHz,DMSO-d6)δ12.93(s,1H),8.81(s,1H),8.17(s,1H),6.94(s,1H),6.71(s,1H),5.76-5.64( m,2H),4.79(s,2H),4.60(d,J=6.9Hz,2H),3.90(s,3H),3.50(s,2H),2.19(s,3H),1.43-1.28(m,12H). LC-MS(M+H) + =498.2.

[0152] Step 5: (E)-3-(4-((tert-Butoxycarbonyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxylic acid

[0153]

[0154] To a solution of methyl (E) -3- (4- ((tert-butoxycarbonyl) amino) but-2-ene-1-yl) -2- (1-ethyl -3-methyl -1H- pyrazole -5- formamido) -3H- imidazo [4,5-b] pyridine -6- carboxylate (760 mg, 1.53 mmol) in methanol (30 mL) was added a solution of NaOH (122 mg, 3.07 mmol) in water (3 mL) and the mixture was stirred at 70 ° C overnight. The mixture was neutralized to pH 7 with HCl (1N) and then concentrated. The residue was dissolved in methanol (10 mL) and the solid was filtered out. The filtrate was concentrated to give the title compound (700 mg, 94%). 1 H NMR(400MHz,DMSO-d6)δ13.25(s,1H),12.92(s,1H),8.79(s,1H),8.17(s,1H),6.95(s,1H),6.71(s,1H ),5.73-5.64(m,2H),4.79(s,2H),4.60(d,J=7.0Hz,2H),3.50(s,2H),2.19(s,3H),1.43-1.10(m,12H). LC-MS(M+H) + =484.2.

[0155] Step 6: tert-Butyl (E)-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)carbamate

[0156]

[0157] A mixture of (E)-3-(4-((tert-butoxycarbonyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxylic acid (700 mg, 1.45 mmol), HATU (581 mg, 1.53 mmol), NH4Cl (409 mg, 7.65 mmol) and DIPEA (395 mg, 3.06 mmol) in N,N-dimethylformamide (8 mL) was stirred at room temperature overnight. The mixture was poured into water (80 mL) and extracted with ethyl acetate (80 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated to give the title compound (560 mg, 76%). 1H NMR (400MHz, DMSO-d6) δ12.90(s,1H),8.76(s,1H),8.16(d,J=12.6Hz,2H),7.56(s,1H),6.94(s,1H),6.70( s,1H),5.77-5.64(m,2H),4.79(s,2H),4.61(d,J=6.0Hz,2H),3.51(s,2H),2.19(s,3H),1.44-1.18(m,12H). LC-MS(M+H) + =483.2.

[0158] Step 7: (E)-3-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide yl)-3H-imidazo[4,5-b]pyridine-6-carboxamide hydrochloride

[0159]

[0160] To a solution of tert-butyl (E)-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)carbamate (560 mg, 1.16 mmol) in dichloromethane (10 mL) was added a solution of HCl in 1,4-dioxane (3 mL, 4 M) and the mixture was stirred at room temperature for 2 h. The precipitate was collected by filtration and dried under vacuum to give the title compound (440 mg, 91%). 1 H NMR(400MHz,DMSO-d6)δ8.77(s,1H),8.21 -8.16(m,2H),7.90(s,3H),7.58(s,1H),6.73(s,1H),6.05(s,1H),5.68( s,1H),4.86(s,2H),4.60(d,J=7.4Hz,2H),3.43(s,2H),2.19(s,3H),1.38 -1.36(m,3H). LC-MS(M+H) + =383.2.

[0161] Step 8: (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3-(4-hydroxybut-2-ene-1- yl)-3H-imidazo[4,5-b]pyridine-6-carboxamide

[0162]

[0163] To a suspension of (E)-3-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide hydrochloride (440 mg, 1.05 mmol) in water (5 mL) was added KBr (276 mg, 2.32 mmol) and NaNO2 (160 mg, 2.32 mmol) and the mixture was stirred at 70° C. for 2 h. After cooling to room temperature, the precipitate was collected by filtration and dried to give the title compound (400 mg, 99%). 1 H NMR(400MHz,DMSO-d6)δ8.75(s,1H),8.16(s,2H),7.52(s,1H),6.69(s,1H),5.82 -5.75(m,2H),4.81(s,2H),4.61(s,2H),3.90(s,1H),3.39(s,2H),2.18(s,3H),1.35(s,3H). LC-MS(M+H) + =384.2.

[0164] Step 9: (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)- 3H-Imidazolo[4,5-b]pyridine-6-carboxamide

[0165]

[0166] To a solution of (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-formamido)-3-(4-hydroxybut-2-ene-1-yl)-3H-imidazo[4,5-b]pyridine-6-carboxamide (400mg, 1.04mmol) in THF (30mL), PBr3 solution in dichloromethane (4mL, 1M) was added and the mixture was stirred at room temperature overnight. The mixture was concentrated and the residue was treated with NaHCO3 aqueous solution (1g in 50mL water). The solid was collected by filtration and the filtrate was extracted with ethyl acetate (50mL). The organic layer was merged with the solid, concentrated under vacuum and purified by column chromatography (DCM / MeOH=50 / 1 to 20 / 1) to give the title compound (190mg, 41%). 1 H NMR (400MHz, DMSO-d6) δ12.92(s,1H),8.75(s,1H),8.16(d,J=14.9Hz,2H),7.55(s,1H),6.73(s,1H),6.05(s,1 H), 5.90 (d, J = 6.8Hz, 1H), 4.85 (s, 2H), 4.60 (d, J = 6.5Hz, 2H), 4.13 (d, J = 7.2Hz, 2H), 2.18 (s, 3H), 1.36 (s, 3H). LC-MS(M+H) +=446.1,448.1.

[0167] Step 10: 1-ethyl-3-methyl-1H-pyrazole and 1-ethyl-5-methyl-1H-pyrazole

[0168]

[0169] To a solution of 3-methyl-1H-pyrazole (50 g, 609 mmol) in DMF (300 mL) was added NaH (60%, 26.8 g, 670 mmol) in portions. The mixture was stirred at 0 °C for 30 min. EtI (105 g, 673 mmol) was added to the mixture. The mixture was stirred at room temperature overnight. The mixture was diluted with water (1000 mL) and extracted with EtOAc (800 mL x 3). The combined organic layers were washed with brine (1000 mL x 3), dried over Na2SO4, filtered and concentrated to give the title compound (62 g, 93%) as a mixture. LC-MS (M+H) + =111.1.

[0170] Step 11: (1-ethyl-3-methyl-1H-pyrazol-5-yl)boronic acid

[0171]

[0172] To a solution of a mixture of 1-ethyl-3-methyl-1H-pyrazole and 1-ethyl-5-methyl-1H-pyrazole (3.5 g, 31.5 mmol) in THF (50 mL) was added n-BuLi (2.5 M, 4.0 mL, 10 mmol) dropwise at -78 °C. The mixture was stirred at -78 °C for 1 h. Triisopropyl borate (1.9 g, 10 mmol) was added to the mixture. The mixture was stirred at -78 °C for 1 h, then warmed to room temperature, diluted with saturated NH4Cl solution (10 mL), and then extracted with THF (30 mL x 2). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was triturated with MTBE (50 mL). The solid was collected by filtration and dried under vacuum to give the title compound (0.7 g, 14%). LC-MS (M+H) + =155.1.

[0173] Step 12: 3-Bromo-5-fluoro-4-nitrobenzamide

[0174]

[0175] A mixture of methyl 3-bromo-5-fluoro-4-nitrobenzoate (5.0 g, 18 mmol) in ammonium hydroxide (25%, 100 mL) was stirred at room temperature overnight. The solid was collected by filtration, washed with water (50 mL) and dried under vacuum to give the title compound (4.0 g, 84%). LC-MS (M+H) + =262.9,264.9.

[0176] Step 13: tert-Butyl (3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)(methyl)carbamate

[0177]

[0178] To a solution of tert-butyl (3-hydroxypropyl) (methyl) carbamate (7.0 g, 37 mmol) in THF (150 mL) was added NaH (60%, 2.8 g, 70 mmol) in batches. The mixture was stirred at room temperature for 30 min. 3-bromo-5-fluoro-4-nitrobenzamide (8.0 g, 30.4 mmol) was added to the mixture. The mixture was stirred at 60 ° C for 1 h. The mixture was cooled to room temperature, then diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE / EtOAc=1 / 1) to give the title compound (12 g, 92%). LC-MS (M+H) + =432.1.

[0179] Step 14: tert-Butyl (3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl) (methyl 4-(2-amino-4-yl)carbamate

[0180]

[0181] To a mixture of tert-butyl (3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)(methyl)carbamate (12 g, 27.8 mmol) and (2-chloropyrimidin-5-yl)boronic acid (4.8 g, 30.3 mmol) in dioxane (200 mL) and water (40 mL) was added K2CO3 (7.7 g, 55.7 mmol) and Pd(dppf)Cl2 (700 mg, 0.96 mmol). The mixture was stirred at 80°C under N2 for 2 h. The mixture was cooled to room temperature and partitioned between EtOAc (300 mL) and water (200 mL). The organic layer was separated, washed with brine (100 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (PE / EtOAc=1 / 1) to give the title compound (4.5 g, 34%). LC-MS (M+H) + =466.1.

[0182] Step 15: tert-butyl(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-(2-nitrophenoxy)propyl)(methyl)carbamate

[0183]

[0184] To a mixture of tert-butyl(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)(methyl)carbamate (4.5 g, 9.7 mmol) and (1-ethyl-3-methyl-1H-pyrazol-5-yl)boronic acid (1.5 g, 9.7 mmol) in dioxane (50 mL) and water (10 mL) was added KCO (4.0 g, 29 mmol) and Pd(dppf)Cl (350 mg, 0.5 mmol). The mixture was stirred at 90 °C under N for 2 h. The mixture was cooled to room temperature, diluted with water (30 mL), and then extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over NaSO, filtered and concentrated. The residue was purified by silica gel column chromatography (EtOAc) to give the title compound (4.5 g, 86%). LC-MS (M+H) + =540.2.

[0185] Step 16: tert-Butyl (3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine 2-(4,5-b)indol-8-yl)oxy)propyl)(methyl)carbamate

[0186]

[0187] To a mixture of tert-butyl(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)propyl)(methyl)carbamate (4.5 g, 8.3 mmol) in 1,2-dichlorobenzene (40 mL) was added 1,2-bis(diphenylphosphino)ethane (6.3 g, 16 mmol). The mixture was stirred at 160° C. under N2 for 8 h. The mixture was cooled to room temperature and poured into petroleum ether (250 mL), the solid was collected by filtration and purified by silica gel column chromatography (DCM / MeOH=15 / 1) to give the title compound (700 mg, 16%). LC-MS (M+H) + =508.3.

[0188] Step 17: tert-butyl (E)-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl) -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- (1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)(methyl)carbamate

[0189]

[0190] A mixture of tert-butyl (3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)(methyl)carbamate (11 mg, 0.02 mmol), (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide (12 mg, 0.024 mmol) and K2CO3 (6 mg, 0.04 mmol) in N,N-dimethylformamide (0.5 mL) was stirred at 70°C for 2 h. The solvent was evaporated and the residue was purified by preparative TLC (DCM / MeOH=10 / 1, containing 0.1% ammonia water) to give Example 1-AA (6 mg, 34%). 1 H NMR(400MHz,CD3OD)δ9.32(s,1H),8.67(s,1H),8.32(s,1H),8.13(s,1H) ,7.58(s,1H),6.84(s,1H),6.36(s,1H),6.06(s,1H),5.75(s,1H),5.41(s ,2H),4.81(s,2H),4.70(s,2H),4.47(s,2H),4.08(s,2H),2.79(s,3H),2. 27 (s, 3H), 2.09 (s, 3H), 1.92 (s, 2H), 1.40 (s, 9H), 1.27 (d, J = 14.4Hz, 6H). LC-MS(M+H) + =873.4.

[0191] Example 1-AB

[0192] (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo [4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-(methylamino) (propyloxy)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0193]

[0194] To a solution of tert-butyl (E)-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)(methyl)carbamate (6 mg, 0.007 mmol) in dichloromethane (1 mL) was added HCl solution (4 M, 0.5 mL) in 1,4-dioxane and the mixture was stirred for 2 h. The mixture was concentrated and the residue was purified by preparative HPLC to give Example 1-AB (1.0 mg, 18%). 1 H NMR(400MHz,CD3OD)δ9.32(s,1H),8.57(s,1H),8.31(s,1H),8.11(s,1H),7.52(s,1H) ,6.84(s,1H),6.36(s,1H),6.01-5.89(m,1H),5.81-5.72(m,1H),5.40-5.25(m,2H),4 .83-4.69(m,4H),4.52-4.42(m,2H),4.10-4.01(m,2H),2.89-2.79(m,2H),2.58(s,3H ),2.27(s,3H),2.12(s,3H),2.02-1.89(m,2H),1.46-1.36(m,3H),1.31-1.20(m,3H). LC-MS(M+H) + =773.7.

[0195] Example 2-AA

[0196] Tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H- pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazine-1-carboxylate

[0197]

[0198] Step 1: tert-Butyl 4-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)piperazine-1-carboxylate

[0199]

[0200] The title compound (10.0 g, 60%) was prepared in a similar manner to Example 1-AA Step 13 from tert-butyl 4-(3-hydroxypropyl)piperazine-1-carboxylate and 3-bromo-5-fluoro-4-nitrobenzamide. LC-MS (M+H) + =487.1.

[0201] Step 2: tert-Butyl 4-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)piperazine-1-carboxylate

[0202]

[0203] The title compound (3.0 g, 28%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 4-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)piperazine-1-carboxylate and (2-chloropyrimidin-5-yl)boronic acid. LC-MS (M+H) + =521.2.

[0204] Step 3: tert-butyl 4-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-(2-nitrophenoxy)propyl)piperazine-1-carboxylate

[0205]

[0206] The title compound (2.3 g, 65%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 4-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)piperazine-1-carboxylate and (1-ethyl-3-methyl-1H-pyrazol-5-yl)boronic acid. LC-MS (M+H) + =595.3.

[0207] Step 4: tert-butyl 4-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine Idio[4,5-b]indol-8-yl)oxy)propyl)piperazine-1-carboxylate

[0208]

[0209] The title compound (500 mg, 23%) was prepared in a similar manner to Example 1-AA Step 16 from tert-butyl 4-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)propyl)piperazine-1-carboxylate and 1,2-bis(diphenylphosphino)ethane. LC-MS (M+H) + =563.3.

[0210] Step 5: tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl) -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- 9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazine-1-carboxylate

[0211]

[0212] Example 2-AA (280 mg, 34%) was prepared in a manner similar to that in Example 1-AA Step 17 from tert-butyl 4-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazine-1-carboxylate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. 1 H NMR(400MHz,CD3OD)δ9.21(s,1H),8.65(s,1H),8.23(s,1H),8.10(s,1H),7.51(s,1H), 6.75(s,1H),6.31(s,1H),6.04(d,J=14.5Hz,1H),5.68(d,J=15.0Hz,1H),5.31(s,2H), 4.75(s,2H),4.66-4.64(m,2H),4.46-4.44(m,2H),4.09(s,2H),3.49(s,4H),2.69-2.6 1(m,6H),2.22(s,3H),2.06(s,3H),1.97(s,2H),1.47(s,9H),1.35(s,3H),1.24(s,3H). LC-MS(M+H) + =928.5.

[0213] Example 2 - AB-HCl

[0214] (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo [4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-(piperazine-1- (4-(2-(2-(4-(2-((4-((2-( ...

[0215]

[0216] Example 2-AB-HCl (200 mg, 77%) was prepared in a similar manner to Example 1-AB from tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazine-1-carboxylate. 1H NMR(400MHz,CD3OD)δ9.59(s,1H),8.76(s,1H),8.45(s,1H),8.21(s,1H),7.74(s,1H),7.06(s ,1H),6.64(s,1H),6.21(d,J=15.1Hz,1H),5.69(d,J=15.1Hz,1H),5.52-5.45(m,2H),4.90(2H under water peak),4.72-4.62(m,4H),4.36-4.30(m,2H),4.00-3.49(m,10H),2.45-2.34(m,5H),2.27(s,3H),1.48-1.35(m,6H). LC-MS(M+H) + =828.5.

[0217] Example 1-AC

[0218] (E)-8-(3-(2-(aminooxy)-N-methylacetamido)propoxy)-9-(4-(6-carbamoyl-2- (1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2- (1-Ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0219]

[0220] Step 1: tert-butyl (E)-(2-((3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3- methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl) (methyl)amino)-2-oxoethoxy 4-(2-amino-4-yl)carbamate

[0221]

[0222] To a mixture of (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-(methylamino)propoxy)-9H-pyrimido[4,5-b]indole-6-carboxamide (90 mg, 0.116 mmol), 2-(((tert-butoxycarbonyl)amino)oxy)acetic acid (33 mg, 0.175 mmol) and DIEA (30 mg, 0.233 mmol) in DMF (5 mL) was added HATU (66 mg, 0.175 mmol). The mixture was stirred at room temperature for 5 h. The mixture was diluted with water, extracted with DCM / IPA (3 / 1, 60 mL x 2), dried over Na2SO4, filtered and concentrated. The residue was triturated with EtOAc to give the title compound (60 mg, 55%). LC-MS (M+H) + =946.

[0223] Step 2: (E)-8-(3-(2-(aminooxy)-N-methylacetamido)propoxy)-9-(4-(6-carbamoyl)- 2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-ene-1- 1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0224]

[0225] To a mixture of tert-butyl (E)-(2-((3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)(methyl)amino)-2-oxoethoxy)carbamate (60 mg, 0.0635 mmol) in DCM / MeOH (8 mL / 4 mL) was added 4M HCl in dioxane (2.0 mL). The mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give Example 1-AC (18 mg, 33%). 1 HNMR(400MHz,CD3OD)δ9.26(s,1H),8.65(s,1H),8.30-8.23(m,1H),8.11(s,1H),7.55-7.47(m,1 H),6.83-6.76(m,1H),6.40-6.31(m,1H),6.06(d,J=15.6Hz,1H),5.78-5.55(m,1H),5.41-5.35( m,2H),4.82-4.58(m,4H),4.52-4.39(m,2H),4.36-4.30(m,2H),4.16-3.96(m,2H),3.45-3.32(m ,2H),2.88(s,3H),2.25(s,3H),2.15-1.84(m,5H),1.39(t,J=6.8Hz,3H),1.23(t,J=6.8Hz,3H). LC-MS(M+H) + =846.5.

[0226] Example 2-AC

[0227] (E)-8-(3-(4-(2-(aminooxy)acetyl)piperazin-1-yl)propoxy)-9-(4-(6-carbamoyl- 2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)- 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0228]

[0229] Step 1: tert-butyl (E)-(2-(4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl- 3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazin-1-yl)-2-oxoethyl oxy) carbamate

[0230]

[0231] To a mixture of (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-(piperazin-1-yl)propoxy)-9H-pyrimido[4,5-b]indole-6-carboxamide hydrochloride (80 mg, 0.1 mmol), 2-(((tert-butoxycarbonyl)amino)oxy)acetic acid (29 mg, 0.15 mmol) and DIEA (52 mg, 0.4 mmol) in DMF (5 mL) was added HATU (57 mg, 0.15 mmol). The mixture was stirred at room temperature for 15 h. The mixture was diluted with water, extracted with DCM / IPA (3 / 1, 60 mL x 2), dried over Na2SO4, filtered and concentrated. The residue was triturated with EA / PE to give the title compound (90 mg, 90%). LC-MS (M+H) + =1001.

[0232] Step 2: (E)-8-(3-(4-(2-(aminooxy)acetyl)piperazin-1-yl)propoxy)-9-(4-(6-amino Formyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)butane-2- 1-Ethyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0233]

[0234] To a mixture of tert-butyl (E)-(2-(4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperazin-1-yl)-2-oxoethoxy)carbamate (90 mg, 0.09 mmol) in DCM / MeOH (8 mL / 4 mL) was added 4M HCl in dioxane (2.0 mL). The mixture was stirred at room temperature overnight, concentrated to dryness and the residue was purified by preparative HPLC to give Example 2-AC (20 mg, 25%). 1H NMR(400MHz,CD3OD)δ9.31(s,1H),8.68(s,1H),8.29(s,1H),8.14(s,1H),7.55(s,1H), 6.83(s,1H),6.34(s,1H),6.10-5.95(m,1H),5.80-5.65(m,1H),5.40-5.32(m,2H),4.8 2-4.65(m,4H),4.62-4.35(m,4H),4.16-4.04(m,2H),3.65-3.52(m,2H),3.48-3.37(m, 2H),2.55-2.32(m,6H),2.26(s,3H),2.08(s,3H),1.95-1.80(m,2H),1.45-1.20(m,6H). LC-MS(M+H) + =901.5.

[0235] Example 3-AA

[0236] Tert-butyl (E)-(4-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrrolidone) oxadiazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyridin-5-yl) oxazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)butyl)(methyl)carbamate

[0237]

[0238] Step 1: tert-Butyl (4-(3-bromo-5-carbamoyl-2-nitrophenoxy)butyl)(methyl)carbamate

[0239]

[0240] The title compound (4.56 g, 68%) was prepared in a similar manner to Example 1-AA Step 13 from tert-butyl N-(4-hydroxybutyl)-N-methylcarbamate and 3-bromo-5-fluoro-4-nitrobenzamide. LC-MS (M-Boc+H) + =348.0.

[0241] Step 2: tert-butyl(4-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)butyl)(methyl 4-(2-amino-4-yl)carbamate

[0242]

[0243] The title compound (388 mg, 28%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 4-(3-bromo-5-carbamoyl-2-nitrophenoxy)butyl(methyl)carbamate and (2-chloropyrimidin-5-yl)boronic acid. LC-MS (M-Boc+H) + =380.1.

[0244] Step 3: tert-butyl (4-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine-5-yl) 2-nitrophenoxy)butyl)(methyl)carbamate

[0245]

[0246] The title compound (194 mg, 43%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 4-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)butyl(methyl)carbamate and (1-ethyl-3-methyl-1H-pyrazol-5-yl)boronic acid. LC-MS (M+H) + =554.4.

[0247] Step 4: tert-butyl (4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidin [4,5-b]indol-8-yloxy)butyl)(methyl)carbamate

[0248]

[0249] The title compound (100 mg, 6%) was prepared in a manner similar to that in Example 1-AA Step 16 from tert-butyl(4-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)butyl)(methyl)carbamate and 1,2-bis(diphenylphosphino)ethane. 1 H NMR(400MHz,DMSO-d6)12.69(s,1H),9.51(s,1H),8.42(s,1H),8.04(s,1H),7.64(s,1H),7.35(s,1H),6.80(s,1H),4.8 5-4.75(m,2H),4.31-4.24(m,2H),3.32-3.25(m,2H),2.80(s,3H),2.23(s,3H),1.88-1.71(m,4H),1.43-1.31(m,12H). LC-MS(M+H) + =522.4.

[0250] Step 5: tert-butyl (E)-(4-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl) -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- 9H-pyrimido[4,5-b]indol-8-yl)oxy)butyl)(methyl)carbamate

[0251]

[0252] Example 3-AA (120 mg, 68%) was prepared in a manner similar to that in Example 1-AA Step 17 from tert-butyl (4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yloxy)butyl)(methyl)carbamate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. 1H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.46(s,1H),8.66(d,J=1.9Hz,1H),8.41(d,J=1.3Hz,1H),8.13(s,1H),8.09(d,J=1.8Hz ,1H),8.03(s,1H),7.59(s,1H),7.53(s,1H),7.38(s,1H),6.77(d,J=0.4Hz,1H),6.40(s,1H),5.99-5.93(m,1H),5.74(s,1H),5 .25(s,2H),4.70(d,J=5.5Hz,2H),4.60(q,J=7.1Hz,2H),4.45(q,J=7.3Hz,2H),4.09(t,J=6.1Hz,2H),3.07(t,J=6.6Hz,2H),2 .69(s,3H),2.21-2.14(m,3H),2.05(s,3H),1.60-1.50(m,4H),1.32-1.29(m,9H),1.25(t,J=7.1Hz,4H),1.20(t,J=7.1Hz,3H). LC-MS(M+H) + =887.6.

[0253] Example 3-AB-HCl

[0254] (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo [4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(4-(methylamino) (butyloxy)-9H-pyrimido[4,5-b]indole-6-carboxamide hydrochloride

[0255]

[0256] Example 3-AB-HCl (80 mg, 78%) was prepared in a similar manner to Example 1-AB from tert-butyl (E)-(4-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)butyl)(methyl)carbamate. 1H NMR(400MHz,DMSO-d6)δ9.48(s,1H),8.71(s,1H),8.54(s,2H),8.45(s,1H),8.21-8.01(m,3H) ,7.64-7.56(m,2H),7.42(s,1H),6.79(s,1H),6.40(s,1H),5.04-5.94(m,1H),5.75-5.65(m,1 H),5.31-5.22(m,2H),4.77-4.70(m,2H),4.65-4.55(m,2H),4.49-4.39(m,2H),4.18-4.08(m, 2H),2.89-2.79(m,2H),2.19(s,3H),2.10-1.98(m,5H),1.76-1.67(m,5H),1.28-1.16(m,6H). LC-MS(M+H) + =787.5.

[0257] Example 4-AA

[0258] Tert-butyl (E)-3-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H- Pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H- pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)azetidine-1-carboxylate

[0259]

[0260] Step 1: tert-Butyl 3-(2-(3-bromo-5-carbamoyl-2-nitrophenoxy)ethyl)azetidine-1-yl Acid esters

[0261]

[0262] The title compound (6.88 g, 91%) was prepared in a similar manner to Example 1-AA Step 13 from 3-bromo-5-fluoro-4-nitrobenzamide and tert-butyl 3-(2-hydroxyethyl)azetidine-1-carboxylate. LC-MS (M-Boc+H) + =343.9.

[0263] Step 2: tert-Butyl 3-(2-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)ethyl) Heterocyclobutane-1-carboxylate

[0264]

[0265] The title compound (556 mg, 28%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 3-(2-(3-bromo-5-carbamoyl-2-nitrophenoxy)ethyl)azetidine-1-carboxylate and 2-chloropyrimidin-5-ylboronic acid. LC-MS (M-Boc+H) + =378.1.

[0266] Step 3: Tert-butyl 3-(2-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-yl)-2-nitrophenoxy)ethyl)azetidine-1-carboxylate

[0267]

[0268] The title compound (230 mg, 36%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 3-(2-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)ethyl)azetidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazol-5-ylboronic acid. LC-MS (M+H) + =552.3.

[0269] Step 4: Tert-butyl 3-(2-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine Imido[4,5-b]indol-8-yl)oxy)ethyl)azetidine-1-carboxylate

[0270]

[0271] The title compound (101 mg, 6%) was prepared in a similar manner to that in Example 1-AA Step 16 from tert-butyl 3-(2-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)ethyl)azetidine-1-carboxylate. 1 H NMR(300MHz,DMSO-d6)δ12.67(s,1H),9.52(s,1H),8.43(s,1H),8.04(s,1H),7.64(s,1H),7.36(s,1H),6.82(s,1H),4.88-4.75(m,2 H),4.31-4.21(m,2H),4.04-3.93(m,2H),3.72-3.61(m,2H),3.00-2.85(m,1H),2.24(s,3H),2.20-2.05(m,2H),1.44-1.33(m,12H). LC-MS(M+H) + =520.3.

[0272] Step 5: tert-butyl (E)-3-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- (1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)azetidine-1-carboxylate

[0273]

[0274] Example 4-AA (100 mg, 56%) was prepared in a similar manner to Example 1-AA Step 17 from tert-butyl 3-(2-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)azetidine-1-carboxylate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.47(s,1H),8.66(d,J=1.9Hz,1H),8.41(d,J=1.3Hz,1H),8.12(s,1H),8.09(d,J= 1.9Hz,1H),8.02(s,1H),7.59(d,J=1.2Hz,1H),7.53(s,1H),7.39(s,1H),6.77(d,J=0.5Hz,1H),6.41(s,1H),6.03-5.92 (m,1H),5.73-5.68(m,1H),5.24(d,J=4.7Hz,2H),4.72(d,J=5.3Hz,2H),4.60(q,J=6.8Hz,2H),4.51-4.43(m,2H),4.04( t,J=6.3Hz,2H),3.85(s,2H),3.45(s,2H),2.18(s,3H),2.05(s,3H),1.91-1.89(m,2H),1.34(s,9H),1.27-1.19(m,7H). LC-MS(M+H) + =885.6.

[0275] Example 4-AB: (E)-8-(2-(azetidin-3-yl)ethoxy)-9-(4-(6-carbamoyl-2-(1-ethoxy) -3-yl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl 3-Methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0276]

[0277] In a similar manner to Example 1-AB, Example 4-AB (40 mg, 46%) was prepared from tert-butyl (E)-3-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)azetidine-1-carboxylate. 1H NMR(400MHz,DMSO-d6)δ9.49(s,1H),8.57(s,1H),8.43(s,1H),8.30(s,1H),8.08-8.03(m, 3H),7.56(s,1H),7.48-4.35(m,2H),6.84(s,1H),6.35(s,1H),5.90-5.80(m,2H),5.28-5.1 7(m,2H),4.81-4.63(m,4H),4.50-4.40(m,2H),4.05-3.91(m,4H),3.61-3.50(m,2H),2.81 -2.74(m,1H),2.22(s,3H),2.06(s,3H),1.93(s,2H),1.36-1.29(m,3H),1.22-1.15(m,3H). LC-MS(M+H) + =785.7.

[0278] Example 5-AA: Tert-butyl (E)-3-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl- 3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)azetidine-1-carboxylate

[0279]

[0280] Step 1: tert-Butyl 3-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)azetidine-1-yl Acid esters

[0281]

[0282] The title compound (2.82 g, 96%) was prepared in a similar manner to Example 1-AA Step 13 from 3-bromo-5-fluoro-4-nitrobenzamide and tert-butyl 3-(3-hydroxypropyl)azetidine-1-carboxylate. LC-MS (M-Boc+H) + =358.1.

[0283] Step 2: tert-Butyl 3-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)nitrogen Heterocyclobutane-1-carboxylate

[0284]

[0285] The title compound (174 mg, 92%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 3-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)azetidine-1-carboxylate and 2-chloropyrimidin-5-ylboronic acid. LC-MS (M-Boc+H) + =392.0.

[0286] Step 3: tert-butyl 3-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-yl)-2-nitrophenoxy)propyl)azetidine-1-carboxylate

[0287]

[0288] The title compound (171 mg, 86%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 3-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)azetidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazol-5-ylboronic acid. LC-MS (M+H) + =566.3.

[0289] Step 4: tert-butyl 3-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine Idio[4,5-b]indol-8-yl)oxy)propyl)azetidine-1-carboxylate

[0290]

[0291] The title compound was prepared in a similar manner to that in Example 1-AA Step 16 from tert-butyl 3-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)propyl)azetidine-1-carboxylate (99 mg, 7%). 1 H NMR(400MHz,DMSO-d6)δ12.71(s,1H),9.53(s,1H),8.42(s,1H),8.04(s,1H),7.63(s,1H),7.35(s,1H),6.81(s,1H),4.88-4.75(m,2 H),4.29-4.19(m,2H),4.01-3.90(m,2H),3.59-3.49(m,2H),2.66-2.56(m,1H),2.24(s,3H),1.88-1.80(m,4H),1.44-1.33(m,12H). LC-MS(M+H) + =534.3.

[0292] Step 5: tert-butyl (E)-3-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- (1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)azetidine-1-carboxylate

[0293]

[0294] Example 5-AA (140 mg, 77%) was prepared in a manner similar to that in Example AA, Step 17, from tert-butyl 3-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)azetidine-1-carboxylate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.46(s,1H),8.66(s,1H),8.40(s,1H),8.13(s,1H),8. 09(s,1H),8.03(s,1H),7.58(s,1H),7.54(s,1H),7.38(s,1H),6.77(s,1H),6.40(s,1H),5.9 3(s,1H),5.70(s,1H),5.25(s,2H),4.70(s,2H),4.61(s,2H),4.45(s,2H),4.03(s,2H),3.82 (s,2H),2.31(s,2H),2.18(s,3H),2.05(s,3H),1.55(s,4H),1.32(s,9H),1.28-1.18(m,7H). LC-MS(M+H) + =899.7.

[0295] Example 5-AB: (E)-8-(3-(azetidin-3-yl)propoxy)-9-(4-(6-carbamoyl-2-(1-ethyl) -3-yl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl 3-Methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0296]

[0297] In a similar manner to Example 1-AB, Example 5-AB (45 mg, 36%) was prepared from tert-butyl (E)-3-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)azetidine-1-carboxylate. 1H NMR(400MHz,DMSO-d6)δ9.50(s,1H),8.65(s,1H),8.44(s,1H),8.33(s,1H),8.17(s,1H),8.10(s,1H),8.05(s ,1H),7.59(s,1H),7.50(s,1H),7.40(s,1H),6.82(s,1H),6.36(s,1H),5.87(d,J=13.5Hz,1H),5.69(d,J=15. 9Hz,1H),5.25(s,2H),4.76-4.62(m,4H),4.50-4.40(m,2H),4.05-3.96(m,2H),3.93-3.89(m,2H),3.53-3.45 (m,2H),2.68-2.61(m,1H),2.21(s,3H),2.06(s,3H),1.59-1.55(m,4H),1.35-1.29(m,3H),1.21-1.15(m,3H). LC-MS(M+H) + =799.7.

[0298] Example 6-AA: Tert-butyl (E)-4-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl- 3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)piperidine-1-carboxylate

[0299]

[0300] Step 1: tert-Butyl 4-(2-(3-bromo-5-carbamoyl-2-nitrophenoxy)ethyl)piperidine-1-carboxylate

[0301]

[0302] The title compound (5.61 g, 63%) was prepared in a similar manner to Example 1-AA Step 13 from 3-bromo-5-fluoro-4-nitrobenzamide and tert-butyl 4-(2-hydroxyethyl)piperidine-1-carboxylate. LC-MS (M-Boc+H) + =372.0.

[0303] Step 2: tert-Butyl 4-(2-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)ethyl)piperidin Pyridine-1-carboxylate

[0304]

[0305] The title compound (331 mg, 90%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 4-(2-(3-bromo-5-carbamoyl-2-nitrophenoxy)ethyl)piperidine-1-carboxylate and 2-chloropyrimidin-5-ylboronic acid. LC-MS (M-Boc+H) + =406.2.

[0306] Step 3: Tert-butyl 4-(2-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-(2-nitrophenoxy)ethyl)piperidine-1-carboxylate

[0307]

[0308] The title compound (196 mg, 52%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 4-(2-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)ethyl)piperidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazol-5-ylboronic acid. LC-MS (M+H) + =580.3.

[0309] Step 4: Tert-butyl 4-(2-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine Idio[4,5-b]indol-8-yl)oxy)ethyl)piperidine-1-carboxylate

[0310]

[0311] The title compound was prepared in a similar manner to that in Example 1-AA Step 16 from tert-butyl 4-(2-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)ethyl)piperidine-1-carboxylate (110 mg, 6%). 1 H NMR(300MHz,DMSO-d6)δ12.67(s,1H),9.50(s,1H),8.40(s,1H),8.02(s,1H),7.63(s,1H),7.34(s,1H),6.79(s,1H),4.85-4.72(m,2 H),4.33-4.23(m,2H),4.00-3.89(m,2H),2.75-2.69(m,2H),2.22(s,3H),1.91-1.67(m,5H),1.47-1.31(m,12H),1.18-1.00(m,2H). LC-MS(M+H) + =548.3.

[0312] Step 5: Tert-butyl (E)-4-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- 9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)piperidine-1-carboxylate

[0313]

[0314] Example 6-AA (140 mg, 71%) was prepared in a similar manner to Example 1-AA Step 17 from tert-butyl 4-(2-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)piperidine-1-carboxylate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. NMR (400MHz, DMSO-d6) δ12.85(s,1H),9.48(s,1H),8.65(d,J=1.8Hz,1H),8.41(d,J=1.2Hz,1H),8.13(s,1H),8 .09(d,J=1.8Hz,1H),8.02(s,1H),7.59(s,1H),7.53(s,1H),7.39(s,1H),6.78(d,J=0.5Hz,1H),6.40(s,1H),6. 00-5.96(m,1H),5.73-5.70(m,1H),5.25(s,2H),4.72(s,2H),4.62(d,J=7.2Hz,2H),4.45(d,J=6.9Hz,2H),4.11 -4.04(m,2H),3.86(s,2H),2.65(s,2H),2.18(s,3H),2.05(s,3H),1.52(s,4H),1.37(s,9H),1.27-1.17(m,9H). LC-MS(M+H) + =913.6.

[0315] Example 6-AB: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)- 3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(2- (Piperidin-4-yl)ethoxy)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0316]

[0317] In a similar manner to Example 1-AB, Example 6-AB (90 mg, 79%) was prepared from tert-butyl (E)-4-(2-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)ethyl)piperidine-1-carboxylate. 1H NMR(400MHz,DMSO-d6)δ9.49(s,1H),8.61(s,1H),8.42(s,1H),8.09-8.04(m,3H),7.6 0(s,1H),7.47(s,1H),7.40(s,1H),6.82(s,1H),6.37(s,1H),6.00-5.75(m,2H),5.32- 5.22(m,2H),4.78-4.62(m,4H),4.53-4.42(m,2H),4.17-4.07(m,2H),3.11-2.99(m,2 H),2.65-2.55(m,2H),2.21(s,3H),2.06(s,3H),1.70-1.50(m,4H),1.37-1.01(m,9H). LC-MS(M+H) + =813.5.

[0318] Example 7-AA: Tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl- 3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl- 3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperidine-1-carboxylate

[0319]

[0320] Step 1: tert-Butyl 4-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)piperidine-1-carboxylate

[0321]

[0322] The title compound (7.56 g, 83%) was prepared in a similar manner to Example 1-AA Step 13 from 3-bromo-5-fluoro-4-nitrobenzamide and tert-butyl 4-(3-hydroxypropyl)piperidine-1-carboxylate. LC-MS (M-Boc+H) + =385.9.

[0323] Step 2: tert-Butyl 4-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)piperidin Pyridine-1-carboxylate

[0324]

[0325] The title compound (358 mg, 37%) was prepared in a similar manner to Example 1-AA Step 14 from tert-butyl 4-(3-(3-bromo-5-carbamoyl-2-nitrophenoxy)propyl)piperidine-1-carboxylate and 2-chloropyrimidin-5-ylboronic acid. LC-MS (M-Boc+H) + =420.1.

[0326] Step 3: Tert-butyl 4-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidine- 5-(2-nitrophenoxy)propyl)piperidine-1-carboxylate

[0327]

[0328] The title compound (195 mg, 48%) was prepared in a similar manner to Example 1-AA Step 15 from tert-butyl 4-(3-(5-carbamoyl-3-(2-chloropyrimidin-5-yl)-2-nitrophenoxy)propyl)piperidine-1-carboxylate and 1-ethyl-3-methyl-1H-pyrazol-5-ylboronic acid. LC-MS (M+H) + =594.4.

[0329] Step 4: tert-butyl 4-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine Idio[4,5-b]indol-8-yl)oxy)propyl)piperidine-1-carboxylate

[0330]

[0331] The title compound was prepared in a manner similar to that in Example 1-AA Step 17 from tert-butyl 4-(3-(5-carbamoyl-3-(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)pyrimidin-5-yl)-2-nitrophenoxy)propyl)piperidine-1-carboxylate (97 mg, 5%). 1 H NMR(300MHz,DMSO-d6)δ12.68(s,1H),9.52(s,1H),8.42(s,1H),8.04(s,1H ),7.63(s,1H),7.35(s,1H),6.81(s,1H),4.87-4.74(m,2H),4.30-4.20(m,2 H),4.00-3.90(m,2H),2.83-2.63(m,2H),2.24(s,3H),1.91-1.83(m,2H),1 .77-1.65(m,2H),1.57-1.47(m,3H),1.45-1.34(m,12H),1.09-0.99(m,2H). LC-MS(M+H) + =562.3.

[0332] Step 5: tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl) -1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3- 9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperidine-1-carboxylate

[0333]

[0334] Example 7-AA (110 mg, 59%) was prepared in a similar manner to Example 1-AA Step 17 from tert-butyl 4-(3-((6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperidine-1-carboxylate and (E)-3-(4-bromobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-carboxamido)-3H-imidazo[4,5-b]pyridine-6-carboxamide. 1 H NMR(400MHz,DMSO-d6)δ12.86(s,1H),9.47(s,1H),8.66(s,1H),8.40(s,1H),8.14(s,1H),8.10(s ,1H),8.02(s,1H),7.55(s,2H),7.38(s,1H),6.79(s,1H),6.39(s,1H),5.94(s,1H),5.72(s,1H),5 .24(s,2H),4.70(s,2H),4.65(s,2H),4.45(s,2H),3.98(s,2H),3.83(s,2H),2.53(S,2H),2.18(s, 3H), 2.04 (s, 3H), 1.60-1.44 (m, 4H), 1.36 (S, 9H), 1.30-1.21 (dd, J = 30.9, 4.6Hz, 8H), 0.86 (s, 3H). LC-MS(M+H) + =927.7.

[0335] Example 7-AB: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)- 3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3- (Piperidin-4-yl)propoxy)-9H-pyrimido[4,5-b]indole-6-carboxamide

[0336]

[0337] Example 7-AB (45 mg, 49%) was prepared in a similar manner to Example 1-AB from tert-butyl (E)-4-(3-((6-carbamoyl-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-3H-imidazo[4,5-b]pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimido[4,5-b]indol-8-yl)oxy)propyl)piperidine-1-carboxylate. 1H NMR(400MHz,DMSO-d6)δ9.50(s,1H),8.66(s,1H),8.43(s,1H),8.33(s,1H),8.17(s,1H),8.11(s,1H),8.05(s,1H),7.58(s,1H),7.51(s,1H ),7.39(s,1H),6.83(s,1H),6.39(s,1H),5.93(d,J=15.3Hz,1H),5.77(d,J=15.5Hz,1H),5.29-5.22(m,2H),4.75-4.65(m,4H),4.50-4.42(m 2H),4.06-4.00(m,2H),3.20-3.12(m,2H),2.75-2.64(m,2H),2.22(s,3H),2.07(s,3H),1.62-1.59(m,4H),1.47-1.01(m,11H). LC-MS(M+H) + =827.7.

[0338] Biological activity

[0339] In THP1-Dual TM STING cell assay in cells

[0340] Material

[0341] THP1-Dual TM The cells are derived from the human THP-1 monocytic cell line by stable integration of two inducible reporter constructs. TM The cells are characterized by the Lucia luciferase gene (a novel secreted luciferase reporter gene) under the control of the ISG54 (interferon stimulated gene) minimal promoter in combination with five interferon (IFN) stimulated response elements. TM The cells also expressed a secreted embryonic alkaline phosphatase (SEAP) reporter gene driven by the IFN-β minimal promoter fused to five copies of the NF-κB consensus transcriptional response element and three copies of the c-Rel binding site. TM The cells allow for the simultaneous study of the NF-κB pathway by monitoring SEAP activity and the interferon regulatory factor (IRF) pathway by assessing Lucia luciferase activity. TM (SEAP detection reagent) and QUANTI-Luc TM Both reporter proteins are easily measured in cell culture supernatants when using the Lucia luciferase assay reagent.

[0342] Different variants of human STING (hSTING) have been identified that affect CDN recognition and signaling:

[0343] R232 (R71-G230-R232-R293): The most common in the human population (about 60%). It is called "wild type" or 232R-RGR allele 2.

[0344] HAQ (H71-A230-R232-Q293): contains three non-synonymous single nucleotide substitutions; R71H, G230A and R293Q. This allele (found in about 20% of the population) is less sensitive to CDN than the "wild type" allele 2.

[0345] Cell maintenance

[0346] Growth medium: RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum (30 min at 56°C), 100 μg / mL Normocin TM ,Pen-Strep(100U / mL-100μg / mL)

[0347] 1. After the cells are recovered (after at least one subculture), they are maintained in growth medium and subcultured. (To maintain the selection pressure, 10 μg / mL blasticidin and 100 μg / mL Zeocin are added every other subculture. TM Add to growth medium.)

[0348] 2. Every 3 days, inoculate 7x10 5 Cells were passaged at 10 cells / ml. The cell concentration was not allowed to exceed 2x10 6 cells / mL.

[0349] Experimental Procedure

[0350] 1. Add 180 μL of cell suspension (approximately 100,000 cells) to each well of a flat-bottom 96-well plate (Costar 3599).

[0351] 2. Compounds were then added at final concentrations ranging from 1 nM to 10 μM in 0.1% DMSO / growth medium via 10 point serial dilutions.

[0352] 3. Incubate the plate at 37°C, 5% CO2 for 24 h.

[0353] 4. Set up the BMG PHERAstar FSX instrument with the following parameters: injection of 50 μL, endpoint measurement, start time of 4 seconds, and read time of 0.1 seconds.

[0354] 5. Add 10 μL THP1-Dual TM The cell culture medium was pipetted into a 96-well white opaque plate (Corning 3903).

[0355] 6. Add 50 μL of QUANTI-Luc assay solution to each well and gently tap the plate several times to mix. Take measurements immediately.

[0356] Table 1. Cellular activity EC of compounds disclosed herein 50 (nM)

[0357]

[0358] It will be appreciated that, if any prior art publication is referred to herein; such reference does not constitute an admission that the publication forms part of the common general knowledge in the art in any country.

[0359] The disclosures of all publications, patents, patent applications, and published patent applications cited herein by acknowledgement are hereby incorporated by reference in their entirety.

[0360] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to one of ordinary skill in the art that certain minor modifications or alterations may be implemented. Therefore, the description and examples should not be construed as limiting the scope of the invention.

Claims

1. A compound having formula (I), or a pharmaceutically acceptable salt thereof, in: R 1 , R 2 , R 6 and R 7 Each independently is -C 1-8 alkyl; R 3 , R 4 and R 8 is hydrogen; R 5 Selected from -R 5a , or -OR 5a ; R 5a Independently selected from -(CH2) n1 -NRR ’ 、-(OCH2CH2) n1 -NRR ’ or R and R' are each independently selected from hydrogen, -C 1-8 Alkyl, -C(=O)OR 9 or -C(=O)-CH2-OR 9 ; R 9 Selected from hydrogen, -C 1-8 Alkyl or -NR 9a R 9b ; R 9a and R 9b Each is hydrogen; Xa and Xb are independently selected from N and CH; n1 is 1, 2, 3, 4, 5, or 6; m1 and m2 are each independently 0 or 1.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 9 Selected from hydrogen, tert-butyl or -NR 9a R 9b .

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, R 5 Selected from -O-(CH2) n1 -NRR ’ or -(OCH2CH2) n1 -NRR ’ .

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 6 and R 7 Each is independently selected from methyl or ethyl.

5. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 6 are each ethyl; and / or R 2 and R 7 Each is a methyl group.

6. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein R 5 Selected from -O-(CH2) n1 -NHR, or -(OCH2CH2) n1 -NHR; n1 is 2, 3 or 4; R is selected from hydrogen, CH3, -C(=O)OR 9 , or -C(=O)-CH2-OR 9 ; R 9 Selected from hydrogen, NH2, or -C(CH3)3.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein R is selected from hydrogen, -CH3, Or -C(=O)OC(CH3)3.

8. A compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein formula (I) is 9. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, wherein formula (II) is Wherein R is selected from CH3, 10. A compound, wherein the compound is or a pharmaceutically acceptable salt thereof.

11. A pharmaceutical composition comprising the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

12. Use of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 11 in the preparation of a medicament for treating a disease affected by STING regulation.

13. The use according to claim 12, wherein the disease is inflammation, allergic and autoimmune diseases, infectious diseases, cancer, or precancerous syndromes.

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