WO2021083383A1 - Nitrogen-containing fused cyclic compound as sting regulator, and preparation method therefor and use thereof - Google Patents

Abstract

Disclosed are a nitrogen-containing fused cyclic compound represented by general formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, a diastereomer, a tautomer, a solvate, a polymorph, or a prodrug thereof, and a preparation method therefor and the use thereof in pharmacy, wherein the definition of each group is as described in the description.

Description

A class of nitrogen-containing fused ring STING regulator compounds, preparation method and use Technical field

The invention belongs to the field of medicinal chemistry, and specifically relates to a class of nitrogen-containing fused ring STING regulator compounds, preparation methods and uses.

Background technique

Interferon gene stimulating protein (STING) is a transmembrane protein, usually in the 152-173 region (dimerization domain) to form a dimer and is in a state of self-inhibition. STING is an important component of the human innate immune system and the body's first line of defense against the invasion of external pathogens such as bacteria and viruses. It plays an important role in maintaining the body's homeostasis, resisting external infections, and preventing tumors and autoimmune diseases.

Studies have shown that the lack of STING activity can trigger tumors or some specific viral infections. For example, the dengue virus protease NS2B3 can degrade STING to hinder the production of IFN-α/β. Therefore, STING activators can be used as vaccine adjuvants or immune activators.

Conversely, excessive activation of STING can cause a variety of serious auto-inflammatory and autoimmune diseases in humans, including rare diseases such as Aicardi-Goutières syndrome (AGS), infantile-onset STING-related vascular disease (SAVI) and systemic erythema Lupus (SLE). In the case of mitochondrial dysfunction, abnormal cGAS/STING activation can induce more common diseases, such as non-alcoholic steatohepatitis (NASH), chronic obstructive pulmonary disease (COPD), age-related macular degeneration (AMD) and Parkinson's Disease.

The important function of STING in the innate immune system and its relevance to various diseases make it a popular target for drug development. A number of pharmaceutical companies are conducting research on agonists and antagonists targeting STING. Cyclic dinucleotide (CDN) compounds are the only agonists discovered so far that can directly activate both murine and human STING protein. Direct injection of CDN compounds into B16 melanoma, CT26 rectal cancer and 4T1 breast cancer masses not only leads to obvious inhibition until the tumor disappears, but also induces persistent antigen-specific T cell immunity of the system, resulting in no injection in other parts of the animal The tumor growth of the drug is also inhibited, causing changes in the microenvironment of a variety of solid tumors, activating effective tumor-induced CD8+ T cells and long-lasting therapeutic effects.

However, the clinical application of CDN compounds for the treatment of anti-tumor, anti-viral, degenerative diseases or immune diseases is still in the very early stage of research, and is limited by the route of administration and the characteristics of druggability, so it must become an effective clinical therapy. There are undoubtedly many obstacles to overcome. Therefore, finding and searching for STING modulators with high activity and good druggability has become a hot field today.

Summary of the invention

One of the technical problems to be solved by the present invention is to provide a novel STING protein regulator for preparing therapeutic drugs for tumors, immune diseases, antiviral and degenerative diseases.

The solutions to the above technical problems are as follows:

A nitrogen-containing heterocyclic compound as shown in the general formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, Polymorphs or prodrugs,

Where:

R _{1 is} selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, 5-10 membered aryl Group or heteroaryl group, acyl group, sulfonyl group, sulfone group, sulfoxide group, etc.;

R _{2 is} selected from amino, amide, C1-C10 alkyl, 5-10 membered aryl or heteroaryl, C3-C6 cycloalkyl or heterocycloalkyl;

M ₁ , M ₂ , M ₃ , M ₄ , M ₅ , M ₆ , and M _{7 are} independently selected from N, CRa, and Ra are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, carbonyl, amide, ester Group, urea, sulfone group, sulfoxide group, sulfonyl group, sulfinyl group, sulfinimide group, C1-C6 alkyl group, C1-C6 alkoxy group, C2-C6 alkenyl group, C2-C6 Alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, 5-10 membered aryl or heteroaryl; or M3=M4, M5=M6 are independently selected from O, S and other hetero atom;

In addition, between any two adjacent Ras mentioned above, a 4- to 10-membered saturated or partially unsaturated ring system can be formed through carbon chains or heteroatoms; or any Ra on M6 and M7 can be combined with any group on R1 A 3-10 membered saturated or partially unsaturated ring system is formed through carbon chains or heteroatoms.

One or more hydrogen atoms on any of the above groups may be substituted by substituents selected from the following group: including but not limited to deuterium, halogen, C1-C8 alkyl; wherein, the heteroaryl includes 1- 3 heteroatoms selected from the group: N, O, P or S, the heterocycloalkyl group contains 1-3 heteroatoms selected from the group: N, O, P or S, the ring The system includes a saturated or partially unsaturated ring system such as a spiro ring, a bridged ring, a condensed ring, and a fused ring. The ring system may be further substituted with a C1-C6 alkyl group, a hydroxyl group, an amino group, a halogen, an alkoxy group, etc.

In some embodiments, the compound of general formula (I) is preferably selected from the compound represented by the following general formula (II), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, or mutual Tautomers, solvates, polymorphs or prodrugs:

Among them, Ra1, Ra2, Ra3, Ra4, Ra5, Ra6, Ra7 are each independently selected from hydrogen, halogen, hydroxyl, amino, cyano, carbonyl, amide, ester, urea, sulfone, sulfoxide, sulfonyl , Sulfinyl group, sulfinimide group, C1-C6 alkyl group, C1-C6 alkoxy group, C2-C6 alkenyl group, C2-C6 alkynyl group, C3-C6 cycloalkyl group, C3 -C6 heterocycloalkyl, 5-10 membered aryl or heteroaryl; and any two adjacent Ra _1-7 can form a 4-10 membered saturation or part through a carbon chain or a heteroatom An unsaturated ring system; or any one of Ra _1-7 can form a 3-10 membered saturated or partially unsaturated ring system with any group on R1 through a carbon chain or a heteroatom; R1, R2 are as defined above.

In some embodiments, it is preferably a compound represented by the following general formula (III), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, or solvent Compounds, polymorphs or prodrugs:

Wherein, R2a, R2b, R2c, Ra4, Ra6, R1' are independently selected from hydrogen, halogen, hydroxyl, amino, cyano, carbonyl, amide, ester, urea, sulfone, and sulfoxide. Group, sulfonyl group, sulfinyl group, sulfinimide group, C1-C6 alkyl group, C1-C6 alkoxy group, C2-C6 alkenyl group, C2-C6 alkynyl group, C3-C6 ring Alkyl, C3-C6 heterocycloalkyl, 5-10 membered aryl or heteroaryl; Ma8 is independently selected from NRa8, O, S(O)p, C(Ra8)q, wherein Ra8 is selected from hydrogen , Halogen, C1-C6 alkyl, p is selected from 0-2, q is selected from 1-2;

And any one or more hydrogens on the above groups can be substituted by groups selected from the following group: -ORm, -NRmRn, -NRmCORn, -CO2Rm, -OCORm, -CONRmRn, -SO2NRmRn, -NRmSO2Rn, -SRm,- SORm, -SO2Rm, -OCONRmRn, -NRpCONRmRn; Rm and Rn are each independently selected from C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyl hydroxy, C1-C6 alkyl alkoxy, C1 -C6 alkylamino or substituted amino, C1-C6 alkyl cyclic amino, C3-C6 cycloalkyl or heterocycloalkyl, 5-8 membered aryl or heteroaryl;

Or Rm and Rn, R2c and R2b can be respectively connected through carbon atoms or heteroatoms to form a 3-12 membered monocyclic or polycyclic alkyl group, a 3-12 membered monocyclic or polycyclic heterocycloalkyl group, and a 3-12 membered A spiro ring or fused ring alkyl group and a 3-12 membered spiro ring or fused ring heterocycloalkyl group;

Wherein, R ₁ , Ra ₄ , and Ra ₆ are as defined above.

The structure shown in Formula III above excludes the following two molecules:

In some preferred embodiments, the compounds of general formulae (I), (II), (III) and (IV), or pharmaceutically acceptable salts thereof, or enantiomers or diastereomers thereof A conformer, tautomer, solvate, polymorph or prodrug, characterized in that the compound includes but is not limited to the following structure:

A method for preparing the compound of general formula I, characterized in that the method comprises steps a-c:

a) reacting a compound of general formula (A) with a propenyl dihalide under alkaline conditions to obtain a compound of general formula (B); and

b) The compound of general formula B is reacted with the compound of general formula R 1'R1”NH under basic conditions to obtain compound of general formula (I); and

c) The compound (I) of the general formula is subjected to a hydrogenation reduction reaction under the catalysis of a transition metal to obtain the compound (I') of the general formula;

Wherein, X is a leaving group such as halogen and sulfonate, and the other groups are as defined above.

Preferably, the steps a), b), and c) are each performed in a solvent, and the solvent is selected from the group consisting of water, methanol, ethanol, isopropanol, butanol, ethylene glycol, and ethylene glycol. Ether, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, toluene, methylene chloride, 1,2-dichloroethane, acetonitrile, N,N-dimethylformamide, N,N-dimethyl Acetamide, dioxane, or a combination thereof.

Preferably, the inorganic base is selected from the group consisting of sodium hydride, potassium hydroxide, sodium acetate, potassium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, carbonic acid Potassium hydrogen, sodium carbonate, sodium hydrogen carbonate, or a combination thereof; the organic base is selected from the group consisting of pyridine, triethylamine, N,N-diisopropylethylamine, 1,8-diazabicyclo [5.4.0] Undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide, lutidine, or a combination thereof.

Another object of the present invention is to provide a medicine and composition for treating or preventing tumors, viral infections, autoimmune diseases and degenerative diseases. The technical solutions to achieve the above objectives are as follows:

A pharmaceutical composition for the treatment or prevention of the above-mentioned diseases, which is composed of the nitrogen-containing heterocyclic compound represented by the above-mentioned general formula (I), or a pharmaceutically acceptable salt thereof, or an enantiomer or diastereomer thereof Isomers, tautomers, solvates, polymorphs or prodrugs and a pharmaceutically acceptable carrier.

Another object of the present invention is to provide a use of the above-mentioned compound. The technical solutions to achieve the above objectives are as follows:

The nitrogen-containing heterocyclic compounds represented by the general formula (I), or pharmaceutically acceptable salts thereof, or enantiomers, diastereomers, tautomers, solvates, Polymorphs or prodrugs are used to prepare drugs for the treatment of STING protein-dependent diseases, especially for tumors, viral infections, immune diseases, and inflammatory diseases, and are a new class of therapeutic drugs with a new mechanism of action. The tumors are each independently selected from: non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, gastric cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia , Lymphoma, fibroma, sarcoma, basal cell carcinoma, glioma, kidney cancer, melanoma, bone cancer, thyroid cancer, nasopharyngeal cancer, pancreatic cancer, etc. The immune diseases and inflammatory diseases are independently selected from the rejection of transplanted organs, gout, rhinitis, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, arthritis, allergic dermatitis, shellfish Chett's disease, bullous skin disease, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease, liver cirrhosis, degenerative joint disease, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, Gastritis, nephritis, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease, irritable bowel syndrome, Kawasaki disease, meningitis, multiple sclerosis, myocarditis, myasthenia gravis, mycosis fungoides, muscle Inflammation, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, psoriasis, fibrosis, lupus erythematosus, tissue Transplant rejection, thyroiditis, type I diabetes, urethritis, uveitis, vasculitis, vitiligo, Waldenstrom's macroglobulinemia, etc.

It should be understood that, within the scope of the present invention, the above technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them one by one here.

Detailed ways

After long-term and in-depth research, the inventor prepared a class of compounds with a novel structure as shown in formula I, and found that it has better STING protein binding activity, and the compound is at a lower concentration (as low as ≤1nmol /L), it can specifically bind to the STING protein, and can regulate the release of the STING pathway or inhibit downstream cytokines such as IFN-β, IL 6, TNF, etc., so it can be used to regulate the activity of the STNG pathway and treat related diseases Such as tumor, inflammation, antiviral, etc. Based on the above findings, the inventor completed the present invention.

the term

Unless otherwise defined, all scientific and technological terms herein have the same meanings as commonly understood by those skilled in the art to which the subject of the claims belongs. Unless otherwise stated, all patents, patent applications, and publications cited in this article are incorporated herein by reference in their entirety.

It should be understood that the foregoing brief description and the following detailed description are exemplary and only used for explanation, and do not impose any limitation on the subject of the present invention. In this application, unless specifically stated otherwise, the use of the singular number also includes the plural number. It must be noted that unless the context clearly indicates otherwise, the singular form used in this specification and claims includes the plural form of the thing referred to. It should also be noted that the use of "or" and "or" means "and/or" unless stated otherwise. In addition, the term "including" and other forms such as "including", "including" and "containing" are not limiting.

Definitions of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols.A (2000) and B (2001), Plenum Press, New York). Unless otherwise specified, conventional methods within the technical scope of the art, such as mass spectrometry, NMR, IR, and UV/VIS spectroscopy and pharmacological methods, are used. Unless specific definitions are provided, the terms used in the description of analytical chemistry, synthetic organic chemistry, and pharmaceuticals and medicinal chemistry herein are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and treatment of patients. For example, the manufacturer's instructions for the use of the kit can be used, or the reaction and purification can be carried out in a manner known in the art or the instructions of the present invention. Generally, the above-mentioned techniques and methods can be implemented according to the descriptions in a number of summary and more specific documents cited and discussed in this specification according to conventional methods well-known in the art. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural parts and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes the chemically equivalent substituent obtained when the structural formula is written from right to left. For example, -CH2O- is equivalent to -OCH2-.

The chapter headings used in this article are only for the purpose of organizing the article, and should not be construed as a limitation on the subject. All documents or document parts cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and theses, are incorporated herein by reference in their entirety.

Certain chemical groups defined herein are preceded by simplified symbols to indicate the total number of carbon atoms present in the group. For example, C1-6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the simplified notation does not include the carbons that may be present in the substituents of the group.

In addition to the foregoing, when used in the specification and claims of this application, unless otherwise specified, the following terms have the following meanings.

In this application, the term "halogen" refers to fluorine, chlorine, bromine or iodine; "hydroxyl" refers to the -OH group; "hydroxyalkyl" refers to an alkane as defined below substituted by a hydroxyl (-OH) "Carbonyl" refers to the -C(=O)- group; "nitro" refers to -NO ₂ ; "cyano" refers to -CN; "amino" refers to -NH ₂ ; "substituted amino" Refers to an amino group substituted with one or two alkyl groups, alkylcarbonyl groups, aralkyl groups, and heteroaralkyl groups as defined below, for example, monoalkylamino, dialkylamino, alkylamido, aralkyl Alkylamino, heteroaralkylamino; "carboxy" refers to -COOH.

In this application, as a group or a part of other groups (for example, used in halogen-substituted alkyl groups and other groups), the term "alkyl" means only composed of carbon atoms and hydrogen atoms without unsaturation. A bond, a straight or branched hydrocarbon chain group having, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms and connected to the rest of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2 , 2-Dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl and decyl, etc.

In this application, as a group or a part of other groups, the term "alkenyl" means consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10 One, more preferably 2 to 6) carbon atoms and a straight or branched hydrocarbon chain group connected to the rest of the molecule through a single bond, such as but not limited to vinyl, propenyl, allyl, but- 1-alkenyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl, etc.

In this application, as a group or part of other groups, the term "alkynyl" means consisting only of carbon atoms and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds, having, for example, A straight or branched hydrocarbon chain group having 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and connected to the rest of the molecule by a single bond, such as but not limited to ethynyl , Prop-1-ynyl, but-1-ynyl, pent-1-en-4-ynyl, etc.

In this application, as a group or part of other groups, the term "cycloalkyl" means a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting of only carbon atoms and hydrogen atoms, which may include condensed The ring system, bridged ring system or spiro ring system has 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, and it is saturated or unsaturated and can be passed through any suitable The carbon atom of is connected to the rest of the molecule by a single bond. Unless specifically indicated otherwise in this specification, the carbon atoms in the cycloalkyl group may be optionally oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H- Indenyl, 2,3-indanyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzene And cyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl , Fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2] Octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octyl Hydro-4,7-methylene-1H-indenyl and octahydro-2,5-methylene- and cyclopentadienyl, etc.

In this application, as a group or part of another group, the term "heterocyclic group" means a group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur Stable 3- to 20-membered non-aromatic cyclic group. Unless otherwise specified in this specification, the heterocyclic group may be a monocyclic, bicyclic, tricyclic or more ring system, which may include a fused ring system, a bridged ring system or a spiro ring system; in the heterocyclic group The nitrogen, carbon, or sulfur atoms of may be optionally oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group can be connected to the rest of the molecule via a carbon atom or a heteroatom and through a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl groups as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purpose of the present invention, the heterocyclic group is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. The group is more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]non Alkyl-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptan-2-yl, aza Cyclobutanyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxopentyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, Imidazolidinyl, quinazinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indoline, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl , Phthalimide, etc.

In this application, as a group or part of another group, the term "aryl" means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms (preferably having 6 to 10 carbon atoms). For the purpose of the present invention, the aryl group can be a monocyclic, bicyclic, tricyclic or more cyclic ring system, and can also be fused with the above-defined cycloalkyl or heterocyclic group, provided that the aryl group passes through The atoms on the aromatic ring are connected to the rest of the molecule by a single bond. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthryl, phenanthryl, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-Benzoxazine-3(4H)-one-7-yl and the like.

In this application, the term "arylalkyl" refers to the above-defined alkyl group substituted by the above-defined aryl group.

In this application, as a group or part of another group, the term "heteroaryl" means having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 selected from nitrogen in the ring A 5- to 16-membered conjugated ring system group of heteroatoms of, oxygen and sulfur. Unless specifically indicated otherwise in this specification, heteroaryl groups can be monocyclic, bicyclic, tricyclic or more cyclic ring systems, and can also be fused with cycloalkyl or heterocyclic groups as defined above, provided that the hetero The aryl group is connected to the rest of the molecule via a single bond through an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group can be optionally oxidized; the nitrogen atom can be optionally quaternized. For the purpose of the present invention, the heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably contains 1 to 4 selected heteroatoms. A stable 5- to 10-membered aromatic group from heteroatoms of nitrogen, oxygen, and sulfur, or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, Benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indazinyl, isoindolyl, indazolyl, isoindazolyl , Purinyl, quinolinyl, isoquinolinyl, naphthyl, naphthyridinyl, quinoxalinyl, pterridinyl, carbazolyl, carboline, phenanthridinyl, phenanthrolinyl, acridine Group, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxtriazolyl, cinnolinyl, quinazolinyl, phenylthio, indolizinyl, phenanthroline, Isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thienyl, naphthopyridyl, [1,2,4]triazolo[4 ,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1, 2,4]Triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine Wait.

In this application, the term "heteroarylalkyl" refers to the above-defined alkyl group substituted by the above-defined heteroaryl group.

In this application, "optional" or "optionally" means that the event or condition described later may or may not occur, and the description includes both occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl group" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups.

The terms "part", "structural part", "chemical moiety", "group", and "chemical group" as used herein refer to specific fragments or functional groups in a molecule. The chemical moiety is generally considered to be a chemical entity embedded or attached to a molecule.

"Stereoisomers" refer to compounds that consist of the same atoms and are bonded by the same bonds, but have different three-dimensional structures. The present invention will cover various stereoisomers and mixtures thereof.

When the compound of the present invention contains an olefinic double bond, unless otherwise specified, the compound of the present invention is intended to include E- and Z-geometric isomers.

"Tautomer" refers to an isomer formed by transferring a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the present invention will also be included in the scope of the present invention.

The compound of the present invention or a pharmaceutically acceptable salt thereof may contain one or more chiral carbon atoms, and therefore may produce enantiomers, diastereomers and other stereoisomeric forms. Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as their racemates and optically pure forms. For the preparation of the compounds of the present invention, racemates, diastereomers or enantiomers can be selected as raw materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

Conventional techniques for preparing/separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives using, for example, chiral high performance liquid chromatography) ), for example, see Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; AMStalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3 :341-63,2010; Fumis et al.(eds.),VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED.,Longman Scientific and Technical Ltd.,Essex,1991,809-816;Heller,Acc.Chem .Res.1990,23,128.

In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic acid or an organic acid that can retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include, but are not limited to, formate, acetate, and 2,2-dichloroacetate , Trifluoroacetate, propionate, caproate, caprylate, caprate, undecylenate, glycolate, gluconate, lactate, sebacate, hexanoate Acid salt, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate , Cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate , Alginate, ascorbate, salicylate, 4-aminosalicylate, naphthalene disulfonate, etc. These salts can be prepared by methods known in the art.

"Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include but are not limited to the following salts: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins , Such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, bicyclic Hexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperazine Pyridine, N-ethylpiperidine, polyamine resin, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the art.

"Polymorphs" refer to different solid crystal phases produced by the presence of two or more different molecular arrangements in certain compounds of the present invention in a solid state. Certain compounds of the present invention may exist in more than one crystal form, and the present invention is intended to include various crystal forms and mixtures thereof.

Generally, crystallization will produce solvates of the compounds of the invention. The term "solvate" as used in the present invention refers to an aggregate comprising one or more molecules of the compound of the present invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of the present invention may exist as hydrates, including monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate, etc., and corresponding solvated forms. The compound of the present invention can form a real solvate, but in some cases, it can also retain only the indeterminate water or a mixture of water and a part of the indeterminate solvent. The compound of the present invention can be reacted in a solvent or precipitated or crystallized from the solvent. Solvates of the compounds of the present invention are also included in the scope of the present invention.

The present invention also includes prodrugs of the above-mentioned compounds. In the present application, the term "prodrug" means a compound that can be converted into the biologically active compound of the invention under physiological conditions or through solvolysis. Therefore, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of the compound of the present invention. When administered to an individual in need, the prodrug may not be active, but is converted into the active compound of the invention in the body. The prodrug is usually rapidly transformed in the body to produce the parent compound of the present invention, for example, by hydrolysis in the blood. Prodrug compounds generally provide advantages in solubility, tissue compatibility, or sustained release in mammalian organisms. Prodrugs include known amino protecting groups and carboxyl protecting groups. For specific preparation methods of prodrugs, please refer to Saulnier, M.G., et al., Bioorg. Med. Chem. Lett. 1994, 4, 1985-1990; Greenwald, R.B., et al., J. Med. Chem. 2000, 43,475.

In the present application, "pharmaceutical composition" refers to a preparation of the compound of the present invention and a medium generally accepted in the art for the delivery of a biologically active compound to a mammal (such as a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of the organism, facilitate the absorption of the active ingredients and then exert the biological activity.

As used herein, the term "pharmaceutically acceptable" refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of the present invention, and is relatively non-toxic, that is, the substance can be administered to an individual without causing undesirable biological activity. Reacts or interacts in an undesirable manner with any components included in the composition.

In this application, "pharmaceutically acceptable carriers" include, but are not limited to, any adjuvants, carriers, excipients, glidants, and sweeteners that are approved by relevant government regulatory agencies as acceptable for human or livestock use. , Diluents, preservatives, dyes/colorants, flavors, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

The "tumor" and "disorders related to abnormal cell proliferation" in the present invention include, but are not limited to, leukemia, gastrointestinal stromal tumor, histocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, lung squamous cell carcinoma, Lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, bowel cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, etc. disease.

The terms "preventive", "preventing" and "preventing" as used herein include reducing the likelihood of the occurrence or exacerbation of a disease or condition in a patient.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) Preventing the occurrence of a disease or condition in a mammal, especially when such a mammal is susceptible to the disease or condition, but has not been diagnosed as having the disease or condition;

(ii) To suppress the disease or disease, that is, to curb its development;

(iii) Alleviate the disease or condition, that is, make the state of the disease or condition subside; or

(iv) Alleviate the symptoms caused by the disease or condition.

The term "effective amount", "therapeutically effective amount" or "pharmaceutical effective amount" as used herein refers to at least one agent or compound that is sufficient to relieve one or more symptoms of the disease or condition being treated after administration的量。 The amount. The result can be a reduction and/or alleviation of signs, symptoms or causes, or any other desired changes in the biological system. For example, the "effective amount" for treatment is the amount of the composition containing the compound disclosed herein that is required to provide significant disease relief clinically. Techniques such as dose escalation tests can be used to determine the effective amount suitable for any individual case.

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

The terms "drug combination", "drug combination", "combination drug", "administration of other treatments", "administration of other therapeutic agents" and the like as used herein refer to drug treatments obtained by mixing or combining more than one active ingredient. It includes fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, combination or sequential administration of at least one compound described herein and at least one synergistic agent to a patient in the form of separate entities. These also apply to cocktail therapy, such as the administration of three or more active ingredients.

Those skilled in the art should also understand that in the methods described below, the functional group of the intermediate compound may need to be protected by an appropriate protecting group. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl (e.g. tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl) , Tetrahydropyranyl, benzyl, etc. Suitable protecting groups for amino, amidino and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable sulfhydryl protecting groups include -C(O)-R" (wherein R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable carboxy protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups can be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protective groups is detailed in Greene, T.W. and P.G.M. Wuts, Protective Groups in Organic Synthesis, (1999), 4th Ed., Wiley. The protecting group can also be a polymer resin.

The present invention will be further explained below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples are usually in accordance with conventional conditions or in accordance with the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and parts by weight.

Preparation of Intermediate A

Intermediate A1: 3-(1,3-Dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester

Step 1: Dissolve methyl 4-iodo-3-methoxybenzoate (3.2g, 11.0mmol) and 5-bromo-2-chloropyridine-4-amine (2.5g, 12.1mmol) in N, N -In dimethylformamide (DMF) (30mL), under the protection of nitrogen, add palladium acetate (747.6mg, 3.34mmol), cesium carbonate (14.5g, 44.5mmol) and 4,5-bis(diphenylphosphine) )-9,9-dimethylxanthene (Xantphos) (341.8mg, 0.59mmol), reacted overnight at 115°C. Cooled to room temperature, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain methyl 4-((5-bromo-2-chloropyridin-4-yl)amino)-3-methoxybenzoate (2.9g, white solid ). LC-MS: m/z 371.0/373.0 [M+H] ⁺ .

Step 2: Dissolve 4-((5-bromo-2-chloropyridin-4-yl)amino)-3-methoxybenzoic acid methyl ester (365mg, 0.99mmol) in DMF (10mL) under nitrogen protection , Sodium acetate (533.1 mg, 6.50 mmol) and bis(triphenylphosphine) palladium(II) chloride (Pd(PPh ₃ ) ₂ Cl ₂ ) (140.4 mg, 0.20 mmol) were added, and reacted overnight at 125°C. Cooled to room temperature, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain methyl 3-chloro-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylate (160mg, white solid ). ¹ H NMR (DMSO-d6, 400MHz): δ 12.42 (s, 1H), 9.30 (s, 1H), 8.57 (s, 1H), 7.61 (d, J = 1.2 Hz, 1H), 7.46 (s, 1H), 4.06 (s, 3H), 3.91 (s, 3H). LC-MS: m/z 291.0/293.0 [M+H] ⁺ .

The third step: Combine 3-chloro-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (500mg, 1.72mmol) and 1,3-dimethyl- 5-pyrazole pinacol borate (453.3mg, 2.04mmol) dissolved in dioxane and water (20mL/4mL), add sodium carbonate (720.8mg, 6.8mmol) and tetratriphenylphosphine under nitrogen protection Palladium (Pd(PPh ₃ ) ₄ ) (196.5 mg, 0.17 mmol) was reacted at 90 degrees overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate A1 (160 mg, white solid). ¹ H NMR (DMSO-d6, 400MHz): δ12.37 (s, 1H), 9.53 (d, J = 0.8 Hz, 1H), 8.58 (d, J = 1.2 Hz, 1H), 7.68 (d, J = 0.8Hz, 1H), 7.60 (d, J = 1.2 Hz, 1H), 6.52 (s, 1H), 4.09 (s, 3H), 4.08 (s, 3H), 3.92 (s, 3H), 2.21 (s, 3H). LC-MS: m/z 351.2 [M+H] ⁺ .

Intermediate A2: 3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxy Methyl ester

以1-乙基3-甲基-5-吡唑频哪醇硼酸酯为原料，参照中间体A1的合成方法制备得到目标中间体。 ¹H NMR(DMSO-d _6,400MHz):δ12.36(s,1H),9.53(s,1H),8.58(s,1H),7.67(s,1H),7.61(s,1H),6.49(s,1H),4.55-4.56(m,2H),4.08(s,3H),3.92(s,3H),2.22(s,3H),1.30-1.32(t,J＝7.2Hz,3H)。LC-MS:m/z 365.2[M+H] ⁺。

Using 1-ethyl 3-methyl-5-pyrazole pinacol borate as raw material, the target intermediate was prepared by referring to the synthesis method of intermediate A1. ¹ H NMR(DMSO-d _6, 400MHz): δ12.36(s,1H), 9.53(s,1H), 8.58(s,1H), 7.67(s,1H), 7.61(s,1H), 6.49 (s, 1H), 4.55-4.56 (m, 2H), 4.08 (s, 3H), 3.92 (s, 3H), 2.22 (s, 3H), 1.30-1.32 (t, J=7.2 Hz, 3H). LC-MS: m/z 365.2 [M+H] ⁺ .

Intermediate A3: 3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-(3-morpholinylpropoxy)-5H-pyrido[4,3-b]indole Methyl indole-8-carboxylate

Step 1: Dissolve methyl 3-hydroxy-4-iodobenzoate (9.3g, 33.5mmol) and 3-morpholinopropyl methanesulfonate (15.0g, 67.3mmol) in DMF (100mL), add carbonic acid Potassium (13.8g, 100.0mmol), react at room temperature overnight. After eluting with ethyl acetate, the filtrate and washing liquid were concentrated under reduced pressure, and purified by column chromatography to obtain methyl 4-iodo-3-(3-morpholinopropoxy)benzoate (12.3 g, white solid). ¹ H NMR(DMSO-d6,400MHz): δ7.94(d,J=8.0Hz,1H), 7.40(d,J=1.6Hz,1H), 7.31(dd,J=1.6,8.0Hz,1H) ,4.14(t,J=6.0Hz,2H),3.86(s,3H),3.55-3.58(m,4H),3.30-3.35(m,2H),2.48-2.51(m,4H),1.90-1.93 (m, 2H). LC-MS: m/z 406.1 [M+H] ⁺ .

Step 2: Combine methyl 4-iodo-3-(3-morpholinopropoxy)benzoate (12.3g, 30.4mmol) and 5-bromo-2-chloropyridine-4-amine (6.9g, 33.5mmol) ) Was dissolved in DMF (150 mL), and under the protection of nitrogen, palladium acetate (680.5 mg, 3.04 mmol), cesium carbonate (29.6 g, 90.8 mmol) and Xantphos (1.75 g, 3.04 mmol) were added and reacted at 100°C overnight. Cooled to room temperature, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain methyl 4-((5-bromo-2-chloropyridin-4-yl)amino)-3-(3-morpholinepropoxy)benzoate (12.0 g, pale yellow solid). LC-MS: m/z 484.4/486.3 [M+H] ⁺ .

Step 3: Dissolve methyl 4-((5-bromo-2-chloropyridin-4-yl)amino)-3-(3-morpholinopropoxy)benzoate (12.0g, 24.84mmol) in DMF (100 mL), under the protection of nitrogen, add sodium acetate (8.15 g, 99.36 mmol) and Pd(PPh ₃ ) ₂ Cl ₂ (1.74 g, 2.48 mmol), and react overnight at 120°C. Cooled to room temperature, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain 3-chloro-6-(3-morpholinepropoxy)-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl Esters (6.1 g, white solid). LC-MS: m/z 404.4/406.3 [M+H] ⁺ .

The fourth step: Combine 3-chloro-6-(3-morpholinepropoxy)-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (220mg, 0.55mmol) and 1, 3-Dimethyl-5-pyrazole pinacol borate (169.7mg, 0.76mmol) was dissolved in dioxane and water (20mL/4mL), and sodium carbonate (233.8mg, 2.2mmol) was added under nitrogen protection React with Pd(PPh ₃ ) ₄ (63.6 mg, 0.055 mmol) at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate A3 (120 mg, white solid). ¹ H NMR(DMSO-d6,400MHz): δ12.23(s,1H), 9.54(s,1H), 8.57(s,1H), 7.71(d,J=0.8Hz,1H), 7.59(d, J = 1.2Hz, 1H), 6.51 (s, 1H), 4.28-4.32 (m, 2H), 4.09 (s, 3H), 3.91 (s, 3H), 3.57-3.61 (m, 4H), 2.50-2.60 (m, 2H), 2.41-2.50 (m, 4H), 2.21 (s, 3H), 1.90-2.12 (m, 2H). LC-MS: m/z 464.3 [M+H] ⁺ .

Using commercially available reagents as raw materials, the following intermediates were prepared by referring to the synthesis method of intermediate A3.

Intermediate B preparation

Intermediate B1: 2-(1,3-Dimethyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid methyl ester

Step 1: Dissolve methyl 4-amino-3-methoxybenzoate (6.0g, 33.15mmol) and 2,4-dichloro-5-iodopyrimidine (11.8g, 43.10mmol) in 2-pentanol (100mL), add N,N-diisopropylethylamine (DIEA) (12.8g, 99.22mmol), and react overnight at 130°C. The reaction solution was cooled to room temperature, filtered, and the solid was slurried and purified with methanol to obtain methyl 4-((2-chloro-5-iodopyrimidin-4-yl)amino)-3-methoxybenzoate (9.4g, white solid) . LC-MS: m/z 420.0/422.0 [M+H] ⁺ .

Step 2: Dissolve 4-((2-chloro-5-iodopyrimidin-4-yl)amino)-3-methoxybenzoic acid methyl ester (9.4g, 22.4mmol) in DMF (100mL) and protect with nitrogen Next, sodium acetate (7.4 g, 90.2 mmol) and bis(triphenylphosphine) palladium(II) chloride (Pd(PPh ₃ ) ₂ Cl ₂ ) (1.57 g, 2.24 mmol) were added, and reacted overnight at 120°C. The reaction solution was cooled to room temperature, poured into water to precipitate solids, filtered, and the filter cake was purified by beating with methanol to obtain 2-chloro-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid methyl ester Esters (5.6 g, white solid). LC-MS: m/z 292.1/294.1 [M+H] ⁺ .

The third step: Combine 2-chloro-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid methyl ester (200mg, 0.69mmol) and 1,3-dimethyl- 5-pyrazole pinacol borate (233.3mg, 1.05mmol) dissolved in dioxane and water (20mL/4mL), add sodium carbonate (300.8mg, 2.84mmol) and Pd(PPh ₃ ) under nitrogen protection ₄ (196.5mg, 0.17mmol), react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate B1 (160 mg, white solid). ¹ H-NMR(DMSO-d6,400MHz): δ12.94(s,1H), 9.67(s,1H), 8.57(d,J=1.2Hz,1H), 7.63(d,J=1.2Hz,1H ), 6.81 (s, 1H), 4.27 (s, 3H), 4.06 (s, 3H), 3.92 (s, 3H), 2.22 (s, 3H). LC-MS: m/z 352.2 [M+H] ⁺ .

Intermediate B2: 2-(1-Ethyl 3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid Methyl ester

以1-乙基3-甲基-5-吡唑频哪醇硼酸酯为原料，参照中间体B1的合成方法制备得到目标中间体。 ¹H NMR(DMSO-d6,400MHz):δ12.92(s,1H),9.66(s,1H),8.57(d,J＝0.8Hz,1H),7.63(d,J＝1.2Hz,1H),6.81(s,1H),4.75-4.79(m,2H),4.06(s,3H),3.92(s,3H),2.23(s,3H),1.39(t,J＝7.2Hz,3H)。LC-MS:m/z 366.2[M+H] ⁺。

Using 1-ethyl 3-methyl-5-pyrazole pinacol borate as raw material, the target intermediate was prepared by referring to the synthesis method of intermediate B1. ¹ H NMR (DMSO-d6, 400MHz): δ 12.92 (s, 1H), 9.66 (s, 1H), 8.57 (d, J = 0.8 Hz, 1H), 7.63 (d, J = 1.2 Hz, 1H) , 6.81 (s, 1H), 4.75-4.79 (m, 2H), 4.06 (s, 3H), 3.92 (s, 3H), 2.23 (s, 3H), 1.39 (t, J = 7.2 Hz, 3H). LC-MS: m/z 366.2 [M+H] ⁺ .

Intermediate B3: 2-(1,3-Dimethyl-1H-pyrazol-5-yl)-8-(3-morpholinylpropoxy)-9H-pyrimido[4,5-b]indole Methyl indole-6-carboxylate

The first step: Dissolve methyl 4-amino-3-hydroxybenzoate (80mg, 0.48mmol) and 3-morpholinoprop-1-ol (73mg, 0.50mmol) in tetrahydrofuran (THF) (25mL), add Diisopropyl azodicarboxylate (DIAD) (194 mg, 0.96 mmol) and triphenylphosphine (PPh ₃ ) (180 mg, 0.70 mmol) were reacted overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain methyl 4-amino-3-(3-morpholinopropoxy)benzoate (70 mg, white solid). LC-MS: m/z 295.2 [M+H] ⁺ .

Subsequent operations refer to the synthesis method of intermediate B1 to prepare intermediate B3. ¹ H NMR(DMSO-d6,400MHz): δ12.89(s,1H), 9.65(s,1H), 8.54(s,1H), 7.60(s,1H), 6.81(s,1H), 4.28- 4.31 (m, 5H), 3.91 (s, 3H), 3.59-3.62 (m, 4H), 2.43-2.60 (m, 9H), 1.90-2.10 (m, 2H). LC-MS: m/z 465.2 [M+H] ⁺ .

Intermediate B4: Methyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxypropoxy)-9H-pyrimido[4,5- b]Indole-6-carboxylate

采用中间体B3相同的方法制备得到中间体B4，LC-MS:m/z410.2[M+H] ⁺。

Intermediate B4 was prepared by the same method as Intermediate B3, LC-MS: m/z 410.2 [M+H] ⁺ .

Intermediate B5: Methyl 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-((4-methoxybenzyl)oxo)-9H-pyrimido[ 4,5-b]indole-6-carboxylate

The first step: Dissolve methyl 3-hydroxy-4-nitrobenzoate (33.0g, 167.5mmol) in DMF (300mL), add potassium carbonate (69.4g, 502.5mmol), reduce the temperature to zero, slowly Add p-methoxybenzyl chloride (PMB-Cl) (31.5g, 201.0mmol) dropwise, react at 80°C for 1h. LC-MS showed that the reaction was complete. After filtration, spin dry, add water (300 mL) and ethyl acetate (150 mL) for extraction twice, spin dry, ethyl acetate (100 mL) to be slurried to obtain methyl 3-((4-methoxybenzyl) (Yl)oxo)-4-nitrobenzoate (yellow solid, 49.2 g). LC-MS[M+H]+: m/z 316.2.

Step 2: Dissolve the yellow solid (2.0g, 6.3mmol) of the previous step in methanol and water (20mL/2mL), add iron powder (1.76g, 31.5mmol) and ammonium chloride (1.70g, 31.5mmol), React at 75 degrees overnight. LC-MS showed that the reaction was complete. After filtering through diatomaceous earth twice, it was spin-dried to obtain a white solid, which was slurried with water (20mL) to obtain methyl 4-amino-3-((4-methoxybenzyl)oxo)benzyl Ester (white solid, 1.6 g).

Step 3: Dissolve the white solid compound of the previous step (3.0g, 10.2mmol), 2,4-dichloro-5-iodopyrimidine (5.58g, 20.4mmol) in 2-pentanol (10mL), and then add DIEA (5.22g, 40.8mmol), reacted overnight at 130°C. After filtration, the solid was slurried with methanol to obtain methyl 4-((2-chloro-5-iodopyrimidin-4-yl)amino)-3-((4-methoxybenzyl)oxo)benzoate ( 3.5g).

Step 4: Dissolve the compound (2.0g, 3.8mmol) obtained in the previous step in DMF (30mL), add sodium acetate (2.06g, 15.2mmol), and then add PdCl ₂ (PPh ₃ ) ₂ Cl ₂ (266mg, 0.38) mmol), under the protection of nitrogen, react at 120°C overnight. Add water (60mL) and ethyl acetate (60mL), column chromatography to obtain methyl 2-chloro-8-((4-methoxybenzyl)oxo)-9H-pyrimido[4,5-b ]Indole-6-carboxylate (1.3g) LC-MS[M+H]+: m/z 398.4.

Step 5: Combine the compound (500mg, 1.25mmol) obtained in the previous step with 1-ethyl-3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxide Boran-2-yl)-1H-pyrazole (450.0mg, 1.9mmol) was dissolved in dioxane and water (24mL/4mL), sodium carbonate (530.0mg, 5.0mmol) was added, and Pd(PPh ₃ ) ₄ (144.0mg, 0.125mmol), under the protection of nitrogen, react at 85°C overnight. Concentrated, and separated by column chromatography (EA/PE=1/1) to obtain Intermediate B5 (white solid, 320 mg). LC-MS[M+H]+: m/z 472.2. 1H NMR (400MHz, DMSO-d6): δ9.29 (s, 1H), 8.35 (s, 1H), 7.87-7.92 (m, 1H), 7.49-7.62(m,2H),6.73-7.16(m,4H),5.32(s,2H),4.82(s,2H),3.77(s,3H),1.99(s,3H),1.17-1.35( m,3H).

Intermediate C: 2-(1-ethyl-3-methyl-1Hpyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole-6-carboxylic acid Methyl ester

The first step: the 4-amino-3-iodo-5-methoxybenzoic acid methyl ester (1.0g, 3.3mmol) and (2,6-dichloropyridin-3-yl)boronic acid (1.26g, 6.6mmol) ) Was added to DMF (50mL), under the protection of nitrogen, added palladium acetate (Pd(OAc) ₂ ) (74mg, 0.33mmol), triphenylphosphine (PPh ₃ ) (86mg, 0.33mmol) and triethylamine (TEA) ) (1.0g, 9.9mmol), heat to 85 degrees and react for 5 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain methyl 4-amino-3-(2,6-dichloropyridin-3-yl)-5-methoxybenzoate (0.7g, pale yellow) solid). ¹ H NMR(DMSO-d6,400MHz): δ7.82(d,J=8.0Hz,1H), 7.61(d,J=8.0Hz,1H), 7.35(s,1H), 7.24(s,1H) , 5.53 (s, 2H), 3.87 (s, 3H), 3.76 (s, 3H). LC-MS: m/z 327.0/329.0 [M+H] ⁺ .

Step 2: Dissolve methyl 4-amino-3-(2,6-dichloropyridin-3-yl)-5-methoxybenzoate (420mg, 1.29mmol) in DMF (20mL), add 18- Crown-6 (680mg, 2.57mmol) and sodium hydride (NaH) (60% in mineral oil, 206mg, 5.15mmol), under the protection of nitrogen, heated to 100°C for 2h. The reaction solution was cooled to room temperature, quenched with water (50 mL), extracted three times with ethyl acetate (50 mL), combined the organic phases, dried, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain 2-chloro-8-methoxy-9H -Pyrido[2,3-b]indole-6-carboxylic acid methyl ester (142 mg, white solid). ¹ H NMR(DMSO-d6,400MHz): δ12.62(brs,1H), 8.72(d,J=8.0Hz,1H), 8.53(s,1H), 7.59(s,1H), 7.34(d, J=8.0 Hz, 1H), 4.05 (s, 3H), 3.91 (s, 3H). LC-MS: m/z 291.1/293.1 [M+H] ⁺ .

The third step: Combine 2-chloro-8-methoxy-9H-pyrido[2,3-b]indole-6-carboxylic acid methyl ester (273mg, 0.94mmol) and 1,3-dimethyl- 5-Pyrazole pinacol borate (278mg, 1.18mmol) was dissolved in dioxane and water (30mL/5mL), and sodium carbonate (398mg, 3.75mmol) and tetrakis(triphenylphosphine) were added under nitrogen protection ) Palladium (Pd(PPh ₃ ) ₄ ) (100 mg, 0.09 mmol), react at 85°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to obtain Intermediate C1 (160 mg, white solid). ¹ H NMR(DMSO-d6,400MHz): δ12.48(brs,1H), 8.73(d,J=8.4Hz,1H), 8.54(s,1H), 7.60-7.64(m,2H)6.63(s , 1H), 4.67-4.70 (m, 2H), 4.08 (s, 3H), 3.93 (s, 3H), 2.23 (s, 3H), 1.38 (t, J=7.2 Hz, 3H). LC-MS: m/z 365.4 [M+H] ⁺ .

Using commercially available reagents as raw materials and referring to the synthesis method of intermediate C1, the following intermediates were prepared.

Intermediate D: (S)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carbamoyl)-3-(3-hydroxypropoxy)-3,4-dihydro -5-oxa-1,2a-acenaphthylene-7-carboxamide

^{The intermediate D, LC-MS[M+H] +} :m/z 413.3 was prepared and synthesized by the same method as that of Example 10 in document WO2020006432A1. ¹ H NMR (400MHz, DMSO): δ 12.72 (s, 1H), 7.93 (s, 1H), 7.59 (s, 1H), 7.34 (s, 2H), 6.64 (s, 1H), 4.08-4.66 ( m, 7H), 3.41-3.43 (m, 2H), 1.83 (s, 3H), 1.34-1.59 (m, 7H).

Intermediate E: (E)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carbamoyl)-3-(4-hydroxybutyl-2-en-1-yl) -3H-imidazole[4,5-b]pyridine-6-carboxamide

Step 1: Dissolve compound (E)-2-(4-bromobut-2-en-1-yl)isoindoline-1,3-dione (17g, 60.93mmol) in DMF (200ml) Then added potassium acetate (11.9g, 121.86mmol), protected by nitrogen, added to 80℃, reacted for 16h, cooled to room temperature, poured into 600mL water, and then extracted with ethyl acetate (200mL*2), the organic layer was saturated with Washed with ammonium chloride (500mL), saturated brine (500mL), dried, and passed through the column to obtain (E)-4-(1,3-dioxoisoindolin-2-yl)but-2-ene- 1-yl acetate (white solid, 13.6 g). ¹ H-NMR (400MHz, DMSO-d ₆ ): δ8.01-7.77 (m, 4H), 5.88-5.75 (m, 1H), 5.75-5.58 (m, 1H), 4.49 (dd, 2H), 4.20 (dd, J=5.2, 2H), 2.08-1.90 (m, 3H).

Step 2: Dissolve the product from the previous step (13.6g, 52.5mmol) in methanol (1.3L), then add sodium methoxide (0.28g, 5.25mmol), stir at room temperature for 16h, then add 14mL 1N HCl to quench, spin dry After passing through the column, (E)-2-(4-hydroxybut-2-en-1-yl)isoindoline-1,3-dione (white solid, 10.6g) was obtained. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ8.29-7.61(m,4H), 5.66(q,2H), 4.71(t,1H), 4.30-4.00(m,2H), 3.89(dd ,2H).

Step 3: Dissolve the product from the previous step (10.6g, 48.85mmol) in dichloromethane (150mL), then add imidazole (6.65g, 97.7mmol) and N,N-dimethylaminopyridine (DMAP) (1.2 g,9.77mmol), stir for 5min, add tert-butyldiphenylchlorosilane (14mL, 53.37mmol) at room temperature, add 200mL of water, stir for 4h, separate the layers, dry the organic layer, spin dry, and pass the column to obtain (E)-2-(4-((tert-butyldiphenylsilyl)oxy)but-2-en-1-yl)isoindoline-1,3-dione (white solid, 21 g). ¹ H-NMR (400MHz, DMSO-d ₆ ): δ7.99-7.79 (m, 4H), 7.58 (dd, 4H), 7.45-7.28 (m, 6H), 5.78 (dt, 1H), 5.69 (dt , 1H), 4.19 (d, 2H), 4.18-4.13 (m, 2H), 0.97 (s, 9H).

Step 4: Dissolve the product from the previous step (21g, 46.15mmol) in ethanol (500mL), add hydrazine hydrate (content 80%, 5.3ml, 184.6mmol), reflux for 16h, cool, filter, and spin-dry the filtrate. Water (200 mL) and ethyl acetate (200 mL) were added, and the layers were separated. The organic layer was washed with water, dried, and spin-dried to obtain (E)-4-((tert-butyldiphenylsilyl)oxy)butan-2- En-1-amino (colorless transparent liquid, 15g). ¹ H-NMR (400MHz, DMSO-d6): δ7.64-7.60 (m, 4H), 7.50-7.34 (m, 6H), 5.78 (dt, 1H), 5.70-5.52 (m, 1H), 4.17 ( dd, 2H), 3.15 (dd, 2H), 1.54-1.30 (m, 2H), 1.00 (s, 9H).

Step 5: Dissolve the product from the previous step (3.5g, 16.4mmol) and methyl 6-chloro-5-nitronicotinic acid ester (5.9g, 18.03 mmol) in dioxane (60mL) and keep at room temperature DIEA (8.1mL, 49.2mmol) was added, stirred at room temperature for 3h, the reaction solution was poured into water (200mL), and extracted with ethyl acetate (300mL), the organic layer was washed with saturated brine, dried, spin-dried, and passed through the column to obtain Methyl (E)-6-((4-((tert-butyldiphenylsilyl)oxy)but-2-en-1-yl)amino)-5-nitronicotinate (yellow solid, 6.3 g). ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.09 (t, 1H), 8.86 (dd, 1H), 8.74 (dd, 1H), 7.59 (dd, 4H), 7.47-7.28 (m, 6H) ), 5.88-5.83 (m, 1H), 5.76-5.62 (m, 1H), 4.26 (t, 2H), 4.17 (t, 2H), 3.88 (d, 3H), 1.00 (s, 9H).

Step 6: Dissolve the product from the previous step (1.5g, 0.97mmol) in methanol/tetrahydrofuran/water (25mL/25mL/25mL), add ammonium chloride (1.6g, 29.7mmol) and add zinc powder (1.93g, 29.7mmol), stirred at room temperature for 3h, filtered, the filtrate was poured into water, extracted with ethyl acetate, and then the organic layer was washed with saturated brine, dried, and spin-dried to obtain methyl (E)-5-amino-6-(( 4-((tert-butyldiphenylsilyl)oxy)but-2-en-1-yl)amino)nicotinate (red-brown sticky substance, 1.4g). LC-MS[M+H] ⁺ : m/z 476.2. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ8.03 (t, 1H), 7.61 (dd, 4H), 7.50-7.32 (m, 6H), 7.17 (d, 1H), 6.53 (t, 1H), 5.87 (dt, 1H), 5.72 (dd, 1H), 5.02 (s, 2H), 4.19 (d, 2H), 4.14-4.01 (m , 2H), 3.74 (s, 3H), 0.99 (s, 9H).

Step 7: Dissolve the product from the previous step (200mg, 0.42mmol) in dichloromethane/anhydrous methanol (3mL/3mL), add cyanogen bromide (108mg, 1.0mmol) at room temperature, and then stir at room temperature for 16h. Saturated sodium bicarbonate solution (50 mL) was added to the reaction solution, stirred for 1 h, and extracted with dichloromethane (50 mL). The organic layer was washed with saturated brine, dried, spin-dried, and purified by TLC plate to obtain methyl (E) -2-Amino-3-(4-((tert-butyldiphenylsilyl)oxy)but-2-en-1-yl)-3H-imidazo[4,5-b]pyridine-6-carboxy Ester (white solid, 50 mg). LC-MS[M+H] ⁺ :m/z501.2. ¹ H-NMR(400MHz,DMSO-d ₆ ):δ8.51(d,1H),7.85(d,1H),7.55(dd,4H ), 7.45-7.39 (m, 2H), 7.35 (dd, 4H), 7.17 (s, 2H), 5.88 (m, 1H), 5.64 (m, 1H), 4.75 (d, 2H), 4.15 (d, 2H), 3.81 (m, 3H), 0.96 (s, 9H).

Example

Example 1: (E)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) -6-Methoxy-5H-pyrido[4,3-b]indole-8-amide

Step 1: Dissolve intermediate A1 (190mg, 0.54mmol) and (E)-1,4-dibromobut-2-ene (58.3mg, 0.27mmol) in DMF (10mL), add cesium carbonate (531.5mg) , 1.62mmol) and sodium iodide (162mg, 1.08mmol), react overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 5,5'-(butyl-2-ene-1,4-diyl)(E)-bis(3-(1,3-dimethyl-1H- Pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester) (75 mg, white solid). LC-MS: m/z 753.4 [M+H] ⁺ .

Step 2: Add 5,5'-(butyl-2-ene-1,4-diyl)(E)-bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) )-6-Methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester) (75mg, 0.10mmol) was dissolved in methanol (10mL), and 2N sodium hydroxide (NaOH) was added (5mL), react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, adjusted to pH=3 by 1M dilute hydrochloric acid, solids separated out, filtered and dried to obtain 5,5'-(butyl-2-ene-1,4-diyl) (E)-Bis(3-(1,3-Dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxy Acid) (70 mg, white solid). LC-MS: m/z 725.3 [M+H] ⁺ .

The third step: 5,5'-(butyl-2-ene-1,4-diyl)(E)-bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) )-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid) (30mg, 0.04mmol) was dissolved in DMF (15mL), and ammonium chloride (NH ₄ Cl) was added ( 23mg, 0.4mmol), DIEA (51.6mg, 0.4mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) (30.4mg, 0.08mmol). React overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by preparative chromatography to obtain (E)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyridine) (Azol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide (0.81mg, white solid). ¹ H NMR (CD ₃ OD, 400MHz): δ9 .39(s,2H),8.43(d,J=1.2Hz,2H),7.78(s,2H),7.60(s,2H),6.22(s,2H),5.92(br s,2H),5.33 -5.37 (m, 4H), 3.84 (s, 6H), 3.83 (s, 6H), 2.26 (s, 6H). LC-MS: m/z 723.2 [M+H] ⁺ .

Example 2: (E)-5-(4-(8-carbamoyl-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyridine And [4,3-b]indol-5-yl)butyl-2-en-1-yl)-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6- Methoxy-N,N-dimethyl-5H-pyrido[4,3-b]indole-8-amide

在制备实施例1过程中，因副反应同时分离得到目标化合物。 ¹H NMR(CD ₃OD,400MHz):δ9.49(s,1H),9.45(s,1H),8.45(d,J＝0.8Hz,1H),8.01(s,1H),7.89,7.90(ss,2H),7.61(s,1H),7.19(s,1H),6.33,6.34(ss,2H),5.92(br s,2H),5.38(br s,4H),3.78-3.86(m,12H),3.18(br s,3H),3.09(br s,3H),2.28,2,29(ss,6H)。LC-MS:m/z 751.2[M+H] ⁺。

During the preparation of Example 1, the target compound was simultaneously separated due to side reactions. ¹ H NMR (CD ₃ OD, 400MHz): δ9.49 (s, 1H), 9.45 (s, 1H), 8.45 (d, J = 0.8 Hz, 1H), 8.01 (s, 1H), 7.89, 7.90 ( ss, 2H), 7.61 (s, 1H), 7.19 (s, 1H), 6.33, 6.34 (ss, 2H), 5.92 (br s, 2H), 5.38 (br s, 4H), 3.78-3.86 (m, 12H), 3.18 (br s, 3H), 3.09 (br s, 3H), 2.28, 2, 29 (ss, 6H). LC-MS: m/z 751.2 [M+H] ⁺ .

Example 3: (E)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) -6-Methoxy-N,N-dimethyl-5H-pyrido[4,3-b]indole-8-amide

在制备实施例1过程中，因副反应同时分离得到目标化合物。 ¹H NMR(CD ₃OD,400MHz):δ9.50(s,2H),8.01(d,J＝1.2Hz,2H),7.91(s,2H),7.20(s,2H),6.35(s,2H),5.93(br s,2H),5.39(br s,4H),3.83(s,6H),3.80(s,6H),3.18(br s,6H),3.09(br s,6H),2.29(s,6H)。LC-MS:m/z 779.2[M+H] ⁺。

During the preparation of Example 1, the target compound was simultaneously separated due to side reactions. ¹ H NMR (CD ₃ OD, 400MHz): δ9.50 (s, 2H), 8.01 (d, J = 1.2Hz, 2H), 7.91 (s, 2H), 7.20 (s, 2H), 6.35 (s, 2H), 5.93 (br s, 2H), 5.39 (br s, 4H), 3.83 (s, 6H), 3.80 (s, 6H), 3.18 (br s, 6H), 3.09 (br s, 6H), 2.29 (s,6H). LC-MS: m/z 779.2 [M+H] ⁺ .

Example 4: (E)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1-ethyl-3-methyl-1H-pyrazole-5- Yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide

Step 1: Dissolve Intermediate A2 (500mg, 1.37mmol) in methanol (20mL), add 2N NaOH (5mL), and react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, 1M dilute hydrochloric acid was adjusted to pH=3, solids precipitated, filtered and dried to obtain 3-(1-ethyl-3-methyl-1H-pyrazole-5- Yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid (420 mg, white solid). LC-MS: m/z 351.1 [M+H] ⁺ .

Step 2: Add 3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8- Carboxylic acid (288mg, 0.82mmol) was dissolved in DMF (6mL), ammonium chloride (NH ₄ Cl) (440mg, 8.2mmol), DIEA (529mg, 4.1mmol) and HATU (935mg, 2.46mmol) were added and stirred at room temperature for 2 hour. The reaction solution was added with water (30mL), and extracted with ethyl acetate (15mL) for 6 times. The organic phases were combined and concentrated to about 3mL. A solid precipitated out. Filtered and dried to obtain 3-(1-ethyl-3-methyl-1H-pyridine). (Azol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide (159 mg, white solid). LC-MS: m/z 350.2 [M+H] ⁺ .

The third step: Add 3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8- Amide (50mg, 0.14mmol) and (E)-1,4-dibromobut-2-ene (15mg, 0.071mmol) were dissolved in DMF (8mL), cesium carbonate (93mg, 0.29mmol) and sodium iodide ( 24mg, 0.16mmol), stirred at room temperature for 3 hours. Concentrated under reduced pressure and purified by preparative chromatography to obtain (E)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1-ethyl-3-methyl-1H-pyridine) (Azol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide (4.5mg, white solid). ¹ H NMR (DMSO-d6,400MHz): δ9 .32(s,2H),8.45(s,2H),8.02(brs,2H),7.76(s,2H),7.57(s,2H),7.36(brs,2H),6.10(s,2H), 5.91 (br s, 2H), 5.25 (brs, 4H), 4.45-4.50 (m, 4H), 3.82 (s, 6H), 2.14 (s, 6H), 1.23 (t, J=7.2 Hz, 6H). MS (ESI): m/z=751.2 [M+H].

Using Intermediate A1 as a raw material, referring to the synthesis methods of Example 1 and Example 4, the following example compounds were prepared.

Example 7: (Z)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) -6-Methoxy-5H-pyrido[4,3-b]indole-8-amide

Step 1: Dissolve intermediate A1 (190mg, 0.54mmol) and (Z)-1,4-dichlorobut-2-ene (58.3mg, 0.27mmol) in DMF (10mL), add cesium carbonate (531.5mg) , 1.62mmol) and sodium iodide (162mg, 1.08mmol), react overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 5,5'-(butyl-2-ene-1,4-diyl)(Z)-bis(3-(1,3-dimethyl-1H- Pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester) (75 mg, white solid). LC-MS: m/z 753.2 [M+H] ⁺ .

Step 2: Add 5,5'-(butyl-2-ene-1,4-diyl)(Z)-bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) )-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester) (75mg, 0.10mmol) was dissolved in methanol (10mL), 2N NaOH (5mL) was added, 100 Reaction overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, adjusted to pH=3 by 1M dilute hydrochloric acid, solids separated out, filtered and dried to obtain 5,5'-(butyl-2-ene-1,4-diyl) (Z)-Bis(3-(1,3-Dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxy Acid) (70 mg, white solid). LC-MS: m/z 725.2 [M+H] ⁺ .

The third step: 5,5'-(butyl-2-ene-1,4-diyl)(Z)-bis(3-(1,3-dimethyl-1H-pyrazol-5-yl) )-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid) (30mg, 0.04mmol) was dissolved in DMF (15mL), and NH ₄ Cl (23mg, 0.4mmol) was added , DIEA (51.6 mg, 0.4 mmol) and HATU (30.4 mg, 0.08 mmol). React overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by preparative chromatography to obtain (Z)-5,5'-(butyl-2-ene-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyridine) (Azol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide (8.5 mg, white solid). ¹ H NMR (DMSO-d6, 400MHz): δ9 .44(s,2H),8.54(s,2H),8.12(brs,2H),8.01(s,2H),7.69(s,2H),7.40(brs,2H),6.48(s,2H), 5.69 (brs, 6H), 4.10 (s, 6H), 4.04 (s, 6H), 2.15 (s, 6H). LC-MS: m/z 723.2 [M+H] ⁺ .

Example 8: 5,5'-(butyl-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H -Pyrido[4,3-b]indole-8-amide

Step 1: Dissolve Intermediate A1 (500mg, 1.43mmol) in methanol (20mL), add 2N NaOH (5mL), and react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, adjusted to pH=3 by 1M dilute hydrochloric acid, solids separated out, filtered and dried to obtain 3-(1,3-dimethyl-1H-pyrazol-5-yl) -6-Methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid (430 mg, white solid). LC-MS: m/z 337.1 [M+H] ⁺ .

Step 2: Add 3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid (430 mg, 1.28 mmol) was dissolved in DMF (25 mL), and NH ₄ Cl (728 mg, 13.7 mmol), DIEA (1.6 g, 12.4 mmol) and HATU (728.4 mg, 1.92 mmol) were added. React overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole Dole-8-amide (330 mg, white solid). LC-MS: m/z 336.2 [M+H] ⁺ .

The third step: the 3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-amide ( 40mg, 0.12mmol) and 1,4-dibromobutane (12.8mg, 0.06mmol) were dissolved in DMF (10mL), cesium carbonate (118mg, 0.36mmol) and sodium iodide (60mg, 0.40mmol) were added and reacted at room temperature overnight. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 5,5'-(butyl-1,4-diyl)bis(3-(1,3-dimethyl-1H-pyrazol-5-yl)- 6-Methoxy-5H-pyrido[4,3-b]indole-8-amide (28.5mg, white solid). ¹ H NMR (CD ₃ OD, 400MHz): δ 9.26 (s, 2H) , 8.27(s, 2H), 7.94(s, 2H), 7.47(s, 2H), 6.56(s, 2H), 4.69(brs, 4H), 3.95(s, 6H), 3.82(s, 6H), 2.33 (s, 6H), 1.97 (brs, 4H). LC-MS: m/z 725.6 [M+H] ⁺ .

Example 9: (E)-9,9'-(butyl-2-ene-1,4-diyl)bis(2-(1,3-dimethyl-1H-pyrazol-5-yl) -8-Methoxy-9H-pyrimido[4,5-b]indole-6-amide

Step 1: Dissolve Intermediate B1 (160 mg, 0.46 mmol) in methanol (10 mL), add 2N NaOH (5 mL), and react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, adjusted to pH=3 by 1M dilute hydrochloric acid, solids separated out, filtered and dried to obtain 2-(1,3-dimethyl-1H-pyrazol-5-yl) -8-Methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid (120 mg, white solid). LC-MS: m/z 338.3 [M+H] ⁺ .

The second step: the 2-(1,3-dimethyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimido[4,5-b]indole-6-carboxylic acid (120 mg, 0.36 mmol) was dissolved in DMF (15 mL), and NH ₄ Cl (199 mg, 3.8 mmol), DIEA (140 mg, 1.08 mmol) and HATU (410.4 mg, 1.08 mmol) were added. React at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and purified by column chromatography to obtain 2-(1,3-dimethyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimido[4,5-b]indole Dole-6-amide (60 mg, white solid). ¹ H NMR (DMSO-d6, 400MHz): δ12.77 (s, 1H), 9.53 (s, 1H), 8.44 (s, 1H), 8.15 (brs, 1H), 7.66 (s, 1H), 7.33 ( brs, 1H), 6.81 (s, 1H), 4.28 (s, 3H), 4.05 (s, 3H), 2.22 (s, 3H). LC-MS: m/z 337.4 [M+H] ⁺ .

The third step: the 2-(1,3-dimethyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimidino[4,5-b]indole-6-amide ( 60mg, 0.18mmol) and (E)-1,4-dibromobut-2-ene (19.3mg, 0.09mmol) were dissolved in DMF (10mL), cesium carbonate (120mg, 037mmol) and sodium iodide (60mg, 0.40mmol), react at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain (E)-9,9'-(butyl-2-ene-1,4-diyl)bis(2-(1,3-dimethyl-1H- Pyrazol-5-yl)-8-methoxy-9H-pyrimido[4,5-b]indole-6-amide (1 mg, white solid). ¹ H NMR (TFA-d, 400MHz): δ11 .67(s,2H),10.60(d,J=0.8Hz,2H),9.80(s,2H),9.37(s,2H),8.11(brs,2H),7.53(brs,4H),6.46( s, 6H), 5.88 (s, 6H), 4.55 (s, 6H). LC-MS: m/z 725.0 [M+H] ⁺ .

Using Intermediate B as a raw material, referring to the synthesis method of Example 9 to prepare the following example compounds.

Example 12: (E)-5-(4-(8-carbamoyl-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-(3-morpholine propoxy) Yl)-5H-pyridine[4,3-b]indol-5-yl)butyl-2-en-1-yl)-3-(1,3-dimethyl-1H-pyrazole-5- Yl)-6-methoxy-5H-pyridine[4,3-b]indole-8-amide

The first step: Intermediate A1 (500mg, 1.43mmol) and (E)-1,4-dibromobut-2-ene (606mg, 2.86mmol) were dissolved in DMF (20mL), and cesium carbonate (1.4g, 4.29mmol) and sodium iodide (429mg, 2.86mmol), react overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain (E)-5-(4-bromobutyl-2-en-1-yl)-3-(1,3-dimethyl-1H-pyrazole-5) -Yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (275 mg, white solid). ¹ H NMR(DMSO-d6,400MHz): δ9.56(s,1H), 8.60(s,1H), 8.03(s,1H), 7.63(s,1H), 6.66(s,1H), 5.95 6.23 (m, 2H), 5.37-5.38 (m, 2H), 4.05-4.16 (m, 8H), 3.93 (s, 3H), 2.22 (s, 3H). LC-MS: m/z 483.1 [M+H] ⁺ .

The second step: (E)-5-(4-bromobutyl-2-en-1-yl)-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6- Methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (275mg, 0.57mmol) and intermediate A3 (264mg, 0.57mmol) were dissolved in DMF (15mL), and cesium carbonate was added (372mg, 1.14mmol) and sodium iodide (86mg, 0.57mmol) at room temperature overnight. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain (E)-3-(1,3-dimethyl-1H-pyrazol-5-yl)-5-(4-(3-(1,3-di Methyl-1H-pyrazol-5-yl)-6-methoxy-8-(methoxycarbonyl)-5H-pyrido[4,3-b]indol-5-yl)butyl-2- En-1-yl)-6-(3-morpholinpropoxy)-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (120 mg, white solid). LC-MS: m/z 866.4 [M+H] ⁺ .

The third step: (E)-3-(1,3-dimethyl-1H-pyrazol-5-yl)-5-(4-(3-(1,3-dimethyl-1H-pyridine) (Azol-5-yl)-6-methoxy-8-(methoxycarbonyl)-5H-pyrido[4,3-b]indol-5-yl)butyl-2-en-1-yl) -6-(3-Morpholinepropoxy)-5H-pyrido[4,3-b]indole-8-carboxylic acid methyl ester (120mg, 0.14mmol) was dissolved in methanol (10mL), and 2N NaOH( 5mL), react at 100°C overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove most of the organic solvents, 1M dilute hydrochloric acid was adjusted to pH=3, solids precipitated, filtered and dried to obtain (E)-5-(4-(8-carboxy-3-(1,3) -Dimethyl-1H-pyrazol-5-yl)-6-(3-morpholinopropoxy)-5H-pyrido[4,3-b]indol-5-yl)butyl-2- En-1-yl)-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8- Carboxylic acid (90 mg, white solid). LC-MS: m/z 838.3 [M+H] ⁺ .

The fourth step: (E)-5-(4-(8-carboxy-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-(3-morpholinepropoxy )-5H-pyrido[4,3-b]indol-5-yl)butyl-2-en-1-yl)-3-(1,3-dimethyl-1H-pyrazole-5- Yl)-6-methoxy-5H-pyrido[4,3-b]indole-8-carboxylic acid (90mg, 0.11mmol) dissolved in DMF (15mL), add NH ₄ Cl (58mg, 1.1mmol) , DIEA (206mg, 1.6mmol) and HATU (84mg, 0.22mmol). React overnight at room temperature. The reaction solution was concentrated under reduced pressure and purified by preparative chromatography to obtain (E)-5-(4-(8-carbamoyl-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-( 3-morpholinopropoxy)-5H-pyridine[4,3-b]indol-5-yl)butyl-2-en-1-yl)-3-(1,3-dimethyl-1H -Pyrazol-5-yl)-6-methoxy-5H-pyridine[4,3-b]indole-8-amide (17 mg, white solid). ¹ H NMR (CD ₃ OD, 400MHz): δ9.58 (s, 1H), 9.57 (s, 1H), 8.56 (d, J = 1.2 Hz, 1H), 8.53 (d, J = 1.2 Hz, 1H) ,8.06(s,1H),8.05(s,1H),7.74(s,1H),7.69(d,J=1.2Hz,1H),6.58(s,1H),6.54(s,1H),5.73- 5.85(m,2H),5.39-5.44(m,4H),4.05-4.20(m,4H),3.71-3.81(m,11H),3.24-3.44(m,6H),2.34(s,3H), 2.33 (s, 3H), 2.10-2.15 (m, 2H). LC-MS: m/z 836.0 [M+H] ⁺ .

Example 13: (E)-5-(4-(8-carbamoyl-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyridine [4,3-b]Indol-5-yl)butyl-2-en-1-yl)-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6 -Methoxy-5H-pyridine[4,3-b]indole-8-amide

The first step: the 3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-methoxy-5H-pyrido[4,3-b]indole-8- Amide (100mg, 0.29mmol) and (E)-1,4-dibromobut-2-ene (318mg, 1.5mmol) were dissolved in DMF (20mL), cesium carbonate (391mg, 1.2mmol) and sodium iodide ( 50mg, 0.3mmol), stirred at room temperature for 1 hour. The reaction solution was separated by adding water (40 mL) and ethyl acetate (40 mL). The aqueous phase was extracted once with ethyl acetate (40 mL), the organic phases were combined, and concentrated under reduced pressure. The residue was slurried with ethyl acetate (20 mL), filtered, and dried to obtain (E)-9-(4-bromobutyl-2) -En-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole -6-amide (100 mg, yellow solid). LC-MS: m/z 482.3 [M+H] ⁺ .

Step 2: Add (E)-9-(4-bromobutyl-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)- 8-Methoxy-9H-pyrido[2,3-b]indole-6-amide (50mg, 0.1mmol) and 3-(1,3-dimethyl-1H-pyrazol-5-yl) -6-Methoxy-5H-pyrido[4,3-b]indole-8-amide (34mg, 0.1mmol) was dissolved in DMF (20mL), cesium carbonate (131mg, 0.4mmol) and sodium iodide were added (16mg, 0.1mmol), stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and purified by preparative chromatography to obtain (E)-5-(4-(8-carbamoyl-3-(1,3-dimethyl-1H-pyrazol-5-yl)-6-methan Oxy-5H-pyridine[4,3-b]indol-5-yl)butyl-2-en-1-yl)-3-(1-ethyl-3-methyl-1H-pyrazole- 5-yl)-6-methoxy-5H-pyridine[4,3-b]indole-8-amide (5.6 mg, white solid). ¹ H NMR (CD ₃ OD, 400MHz): δ9.45 (s, 1H), 9.43 (s, 1H), 8.46 (d, J = 1.2 Hz, 1H), 8.44 (d, J = 1.2 Hz, 1H) , 7.87(s, 1H), 7.81(s, 1H), 7.63(s, 1H), 7.61(s, 1H), 6.30(s, 1H), 6.24(s, 1H), 5.93(br s, 2H) ,5.38(brs,4H),4.24-4.26(m,2H),3.81-3.83(m,9H),2.29(s,3H),2.28(s,3H),1.23(t,J=7.2Hz,3H ). LC-MS: m/z 737.8 [M+H] ⁺ .

Using intermediates A and B as raw materials, referring to the synthesis methods of Example 12 and Example 13, the following example compounds were prepared.

Example 19: (S)-3-(3-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H -Pyrimido[4,5-b]indol-9-yl)propyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-3,4-bis Hydrogen-5-oxa-1,2a-acenaphthylene-7-amide

Prepared by the same method as in Example 12 to obtain the target compound (white solid, 3.1 mg), MS (ESI): m/z=745.4 [M+H]. ¹ H NMR (400MHz, MeOD-d ₄ ): δ9.13 (s, 1H), 8.21 (s, 1H), 7.44 (s, 1H), 7.36 (s, 1H), 7.23 (s, 1H), 6.78 (s, 1H), 6.31 (s, 1H), 5.34 (s, 2H), 4.54-4.80 (m, 6H), 4.09-4.11 (m, 1H), 3.79 (s, 3H), 2.27 (s, 3H) ), 2.12 (s, 3H), 1.20-1.59 (m, 10H).

Example 20: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-3H-imidazole [4, 5-b)pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy- 9H-pyrimido[4,5-b]indole-6-amide

Example 20 (off-white solid, 2.3 mg) was prepared using the same method as Example 12, MS (ESI): m/z=716.1 [M+H].

Example 21: (S)-3-(3-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholine Propoxy)-9H-pyrimido[4,5-b]indol-9-yl)propyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide) -3,4-Dihydro-5-oxa-1,2a-acenaphthylene-7-amide

采用实施例12相同的方法制备得到实施例21(白色固体，5.2mg),MS(ESI)：m/z＝857.9[M+H]。

Example 21 (white solid, 5.2 mg) was prepared using the same method as Example 12, MS (ESI): m/z=857.9 [M+H].

Example 22: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-3H-imidazole [4, 5-b)pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morphine (Alpinoxy)-9H-pyrimido[4,5-b]indole-6-amide

采用实施例12相同的方法制备得到实施例22(白色固体，2.5mg),MS(ESI)：m/z＝829.1[M+H]。

Example 22 (white solid, 2.5 mg) was prepared using the same method as Example 12, MS (ESI): m/z=829.1 [M+H].

Example 23: (S)-3-(3-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxypropyl) Oxy)-9H-pyrimido[4,5-b]indol-9-yl)propyl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)- 3,4-Dihydro-5-oxa-1,2a-acenaphthylene-7-amide

采用实施例12相同的方法制备得到实施例23(白色固体，8mg)，MS(ESI)：m/z＝789.2[M+H]。

Example 23 (white solid, 8 mg) was prepared using the same method as Example 12, MS (ESI): m/z=789.2 [M+H].

Example 24: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamide)-3H-imidazole [4, 5-b)pyridin-3-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxy Propoxy)-9H-pyrimido[4,5-b]indole-6-amide

采用实施例12相同的方法制备得到实施例(白色固体，3.2mg)，MS(ESI)：m/z＝760.2[M+H]。

The Example (white solid, 3.2 mg) was prepared using the same method as Example 12. MS (ESI): m/z=760.2 [M+H].

Example 25: (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-8-(3-(difluoromethoxy)propoxy)-2- (1-Ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-carboxamide

The first step: The compound 3-((4-methoxybenzyl)oxo)propyl-1-ol (1.0g, 5.10mmol) was dissolved in acetonitrile (40mL), and CuI (193.8mg, 1.02 mmol), the temperature was raised to 70°C and 2,2-difluoro-2-(fluorosulfonyl fluoride)acetic acid (1.81g, 10.2mmol) was slowly added, and the reaction was carried out at room temperature for 2 hours. Wash 3 times with ethyl acetate/water (40/40mL), spin dry and mix the sample, use petroleum ether/ethyl acetate (volume ratio PE/EA=10/1) to purify the oily crude product 1-((3 -(Difluoromethoxy)propoxy)methyl)-4-methoxybenzene (300mg). ¹ H-NMR (400MHz, MeOD-d4): δ 7.24 (d, 2H), 6.88 (d, 2H), 6.12 (t, 1H), 4.42 (s, 2H), 3.92 (d, 2H), 3.78 (s, 3H), 3.31 (t, 2H), 1.86-1.90 (m, 2H).

Step 2: Dissolve the crude product of the previous step (10 mg, 0.04 mmol) in methanol (MeOH) (5 mL), add palladium on carbon (2 mg), and react under hydrogen at room temperature overnight. The oily product 3-(difluoromethoxy)propyl-1-ol (4.0 mg) was obtained by filtration and separation.

Step 3: Dissolve the crude product of the previous step (4mg, 0.03mmol) in DCM (10mL), then add TEA (9.1mg, 0.09mmol), cool to 0℃ and slowly add methylsulfonyl chloride (5.2mg, 0.045mmol) , React at room temperature for 2 hours. It was washed 3 times with saturated sodium carbonate (10 mL) and spin-dried to obtain crude oily 3-(difluoromethoxy)propyl methanesulfonate (4 mg), which was directly used in the next reaction.

The fourth step: The crude product of the previous step (5mg, 0.006mmol) and (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-(3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl- 3-Methyl-1H-pyrazol-5-yl)-8-hydroxy-pyrimidine[4,5-b]indole-6-carboxamide (2.5mg, 0.012mmol) dissolved in DMF (5mL), add carbonic acid Cesium (5.9 mg, 0.018 mmol) was reacted at room temperature overnight, extracted with dichloromethane, washed with saturated sodium bicarbonate solution and water, and concentrated to prepare and isolate Example Compound 25 (white solid, 0.6 mg). LC-MS [M+H] ⁺ : m/z 960.4.

Example 26: (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-8-(3-(4,4-difluoropiperidin-1-yl) Propoxy)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-carboxamide

Example 26 (white solid, 6.3 mg) was prepared in the same manner as in Example 25. LC-MS [M+H]+: m/z 1013.4. ¹ H-NMR (400MHz, MeOD-d4): δ9.29 (s, 2H), 8.27 (s, 2H), 7.32 (s, 2H), 6.82 (s, 2H), 5.63 (s, 2H), 5.17 (s, 4H), 4.60-4.66 (m, 6H), 3.68-3.70 (m, 4H), 3.54-3.57 (m, 4H), 2.27-2.32 (m, 4H), 2.25 (s, 6H), 2.20 -2.23 (m, 6H), 1.83-1.87 (m, 4H), 1.46-1.49 (m, 4H), 1.37 (t, 6H).

Example 27: (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-8-(3-(3,3-difluoroazetidine-1 -Yl)propoxy)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-carboxamide

Example 27 (white solid, 5.5 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 985.4. ¹ H-NMR (400MHz, MeOD-d4): δ9.28 (s, 2H), 8.27 (s, 2H), 7.36 (d, 2H), 6.79 (d, 2H), 5.64 (s, 2H), 5.19 (s,4H),4.60-4.64(m,6H),3.74-3.78(m,4H),3.56-3.59(m,4H),4.41-4.47(m,4H),2.33-2.47(m,2H) , 2.26-2.27 (m, 6H), 2.22-2.24 (m, 4H), 1.34-1.38 (m, 4H), 1.30-1.34 (m, 6H).

Example 28: (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrido[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole- 5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indole-6-carboxamide

The first step: the compound 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole-6 -Formamide (100mg, 0.3mmol) and (E)-1,4-dibromobutyl-2-ene (190.0mg, 0.9mmol) were dissolved in DMF (25mL), cesium carbonate (200mg, 0.6mmol) was added, React at room temperature for 1h. Add water (40mL) and ethyl acetate (40mL) to extract once, then beaten with ethyl acetate and petroleum ether (5mL, volume ratio 1/1), filter, and dry to obtain (E)-9-(4-bromobutyl-2) -En-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole -6-Carboxamide (75mg). LC-MS[M+H]+: m/z 484.4.

Step 2: Combine the crude product from the previous step (75mg, 0.16mmol) and 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy)- 9H-pyrimidine[4,5-b]indole-6-carboxamide (74 mg, 0.16 mmol) was dissolved in DMF (5 mL), cesium carbonate (104.3 mg, 0.32 mmol) was added, and the reaction was carried out at room temperature overnight. Column chromatography (volume ratio DCM/MeOH=10/1) was separated to obtain (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazole-5- Yl)-8-methoxy-9H-pyrido[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl -1H-pyrazol-5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indole-6-carboxamide (white solid, 50 mg). LC-MS[M+H]+: m/z 865.6.

Step 3: Dissolve the crude product of the previous step (20mg, 0.02mmol) in DCM (10mL), add boron tribromide (BBr ₃ ) (5mL), and react at 50°C overnight. Quench with methanol and spin dry to obtain crude product (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-hydroxy -9H-pyrido[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5- Yl)-8-(3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indole-6-carboxamide (20 mg). LC-MS [M+H] ⁺ : m/z 851.4.

Step 4: Dissolve the compound from the previous step (20mg, 0.023mmol) and 3-morpholinopropyl-1-ol (13.29mg, 0.046mmol) in DMF (5mL), add cesium carbonate (22.49mg, 0.069mmol), React overnight at room temperature. Dichloromethane extraction, saturated sodium bicarbonate solution and water washing, concentration, preparation and isolation to obtain Example 28 (white solid, 3.7mg). LC-MS [M+H] ⁺ : m/z 978.5. ¹ H-NMR (400MHz, MeOD-d4): δ9.32 (s, 1H), 8.40 (d, 1H), 8.30 (d, 2H), 7.57 (d, 1H), 7.39 (d, 2H), 6.84 (s, 1H), 6.59 (s, 1H), 5.60-5.61 (m, 1H), 5.45-5.49 (m, 1H), 5.21-4.26 (m, 2H), 5.27-5.38 (m, 2H), 4.40 -4.62 (m, 2H), 4.38-4.40 (m, 2H), 3.78-3.79 (m, 12H), 3.08-3.28 (m, 12H), 2.31 (d, 6H), 1.94-1.96 (m, 2H) , 1.81-1.84 (m, 2H), 1.34-1.8. (m, 3H), 1.26-1.36 (m, 3H).

Example 29: (E)-9-(4-(6-amide-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyridine And [2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8 -(3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indole-6-carboxamide

The first step: the 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole-6- Formamide (50mg, 0.14mmol) and (E)-1,4-dibromobutyl-2-ene (148.4mg, 0.7mmol) were dissolved in DMF (20mL), cesium carbonate (136.9mg, 0.42mmol) was added, React at room temperature for 1h. Dilute with water (40mL) and EA (40mL), separate the organic phase, spin dry, beaten with ethyl acetate (20mL), filter, and dry to obtain (E)-9-(4-bromobutyl-2-ene-1-) Yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrido[2,3-b]indole-6-carboxamide (Gray solid, 20 mg). LC-MS [M+H] ⁺ : m/z 484.2.

The second step: the compound (20mg, 0.045mmol) obtained in the previous step and 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy )-9H-pyrimidine[4,5-b]indole-6-carboxamide (19 mg, 0.041 mmol) was dissolved in DMF (20 mL), cesium carbonate (58.7 mg, 0.18 mmol) was added, and the reaction was carried out at room temperature for 1 h. Extracted with dichloromethane, washed with saturated sodium bicarbonate solution and water, concentrated, prepared and isolated the compound of Example 29 (white solid, 35.3 mg). LC-MS [M+H] ⁺ : m/z 865.5. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.48 (s, 1H), 8.51-8.53 (m, 1H), 8.41 (s, 2H), 8.01-8.05 (m, 2H), 7.35-7.60 (m,5H),6.78(s,1H),6.58(s,1H),5.72-5.77(m,2H),5.22(s,4H),4.57-4.59(m,2H),4.22-4.25(m , 2H), 3.93 (s, 2H), 3.77 (s, 3H), 3.45 (s, 4H), 2.11-2.21 (m, 12H), 1.54 (s, 2H), 1.02-1.23 (m, 6H).

Example 30: (E)-9-(4-(8-amide-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-(3-morpholinopropoxy) Yl)-5H-pyrido[4,3-b]indol-5-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole- 5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

The first step: the 2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-methyl Amide (50 mg, 0.14 mmol) and A (148.4 mg, 0.7 mmol) were dissolved in DMF (20 mL), cesium carbonate (136.9 mg, 0.42 mmol) was added, and the reaction was carried out at room temperature for 1 h. Water (40 mL) and ethyl acetate (EA) (40 mL) were added, extracted with ethyl acetate (40 mL), spin-dried, slurried with ethyl acetate (20 mL), filtered, and dried to obtain a gray solid (40 mg). LC-MS [M+H] ⁺ : m/z 485.1.

Step 2: Combine the gray solid of the previous step (40mg, 0.09mmol) with 3-(1-ethyl-3-methyl-1H-pyrazole-5-)-6-(3-morpholinopropoxy)- 5H-pyrido[4,3-b]indole-8-carboxamide (38 mg, 0.81 mmol) was dissolved in DMF (20 mL), cesium carbonate (117.4 mg, 0.36 mmol) was added, and the reaction was carried out at room temperature for 1 h. Extracted with dichloromethane, washed with saturated sodium bicarbonate solution and water, concentrated, prepared and isolated Example 30 (white solid, 35.3 mg). LC-MS [M+H] ⁺ : m/z 865.5. ¹ H-NMR (400MHz, CD ₃ OD): δ9.42 (s, 1H), 9.31 (s, 1H), 8.44 (s, 1H), 8.33 (s, 1H), 7.87 (s, 1H), 7.59 (s, 1H), 7.54(s, 1H), 6.73(s, 1H), 6.29(s, 1H), 5.79(s, 2H), 5.35(s, 2H), 5.28(s, 2H), 4.49- 4.51 (m, 2H), 4.02-4.20 (m, 5H), 3.76 (s, 4H), 3.21-3.30 (m, 8H), 2.05-2.30 (m, 8H), 1.19-1.25 (m, 6H).

Example 31: (E)-9,9'-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-((4-methoxybenzyl)oxo)-9H-pyrimido[4,5-b]indole-6-carboxamide

Intermediate B5 compound (250 mg, 0.54 mmol) and 1,4-dibromobutene (58.0 mg, 0.3 mmol) were dissolved in DMF (25 mL), cesium carbonate (540 mg, 3.0 mmol) was added, and the reaction was carried out at room temperature for 1 h. Water (40 mL) was added to the precipitated solid to be slurried with methanol (10 mL) and DMF (10 mL), filtered, and dried to obtain Example 31 (gray solid, 72 mg). LC-MS[M+H]+: m/z 965.5. 1H-NMR(400MHz,TFA-d6):δ11.58(s,2H),10.47(s,2H),9.72(s,2H),9.38 (s, 2H), 8.14 (s, 2H), 7.51 (s, 4H), 6.93 (s, 4H), 4.55 (s, 6H), 3.63 (s, 6H).

Example 32: (E)-9,9'-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-hydroxy-9H-pyrimido[4,5-b]indole-6-carboxamide)

Example 31 (72 mg, 0.08 mmol) was dissolved in DCM (10 mL), trifluoroacetic acid (TFA) (5 mL) was added, and the reaction was heated at 50°C overnight. It was spin-dried, slurried with methanol (10 mL) and filtered to obtain Example 32 (white solid, 42 mg). 1H-NMR (400MHz, TFA-d6): δ 11.15 (s, 2H), 10.03 (s, 2H), 9.28 (s, 2H), 8.94 (s, 2H), 7.70 (s, 2H), 7.07 ( s, 4H), 6.50 (s, 4H), 4.11 (s, 6H), 3.19 (s, 6H).

Example 33: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-((4-methyl (Oxybenzyl)oxo)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl- 1H-pyrazol-5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

Compound 3a (520.0 mg, 0.88 mmol) and compound 2d (309.5 mg, 0.88 mmol) were dissolved in DMF (20 mL), cesium carbonate (860.6 mg, 2.64 mmol) was added, and the reaction was carried out at room temperature for 2 hours. Water (40 mL) and ethyl acetate (40 mL) were added for extraction once, and then slurried with ethyl acetate and petroleum ether (5 mL, v:v=1:1), filtered, and dried to obtain Example 33 (white solid, 580.0 mg). LC-MS[M+H] ⁺ : m/z 859.6. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.48(s, 2H), 8.45(s, 2H), 8.05(s, 2H), 7.69(s,1H),7.57(s,1H),7.40(s,1H),6.41-7.57(m,2H),7.08-7.11(m,2H),6.75-6.99(m,2H),6.53- 6.63 (m, 2H), 5.69-5.74 (m, 2H), 5.55-5.60 (m, 4H), 5.10 (s, 2H), 4.47-4.57 (m, 4H), 3.59-3.80 (m, 6H), 2.20 (s, 6H), 1.30 (t, 3H), 1.20 (t, J=6.4 Hz, 3H).

Example 34: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-hydroxy-9H-pyrimidine [4,5-b]Indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8- Methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

Example compound 33 (60.0 mg, 0.07 mmol) was dissolved in DCM (10 mL), TFA (5 mL) was added, and the reaction was carried out at room temperature overnight. It was slurried with methanol and dichloromethane (5 mL)=1/1, filtered, and dried to obtain Example 34 (white solid, 50.0 mg). LC-MS[M+H] ⁺ : m/z 739.5. ¹ H NMR (400MHz, TFA-d): δ 11.64 (s, 2H), 10.57 (s, 2H), 9.78 (s, 2H), 9.40 (s, 2H), 8.14 (s, 2H), 7.54 (s, 4H), 6.92-6.95 (m, 4H), 5.89 (s, 3H), 4.57 (s, 6H), 3.60-3.67 (m, 6H) ).

Example 35: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-methoxy Propyloxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-ene-1-ene)-2-(1-ethyl-3-methyl-1H-pyridine (Azol-5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

Example compound 34 (25.0mg, 0.03mmol) and brominated material (9.3mg, 0.06mmol) were dissolved in DMF (10mL), cesium carbonate (29.1mg, 0.09mmol) was added, and the reaction was carried out at room temperature overnight. Preparative chromatographic separation was achieved. Example 35 (white solid, 2.4 mg). LC-MS[M+H] ⁺ : m/z 811.6. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.47(s, 1H), 9.46(s, 1H), 8.43(s, 1H), 8.42 (s, 1H), 8.06 (s, 2H), 7.59 (s, 1H), 7.55 (s, 1H), 7.38-7.41 (m, 2H), 6.75 (s, 1H), 6.72 (s, 1H) , 5.79(s, 2H), 5.20-5.23(m, 4H), 4.49-4.58(m, 4H), 4.00-4.04(m, 2H), 3.77(s, 3H), 3.25-3.28(m, 2H) , 3.11 (s, 3H), 2.19 (s, 3H), 2.16 (s, 3H), 1.71-1.74 (m, 2H), 1.20 (t, 3H), 1.15 (t, 3H).

Example 36: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-((4-( (Trifluoromethyl)benzyl)oxo)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3- Methyl-1H-pyrazol-5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

Example compound 34 (25.0mg, 0.03mmol) and p-trifluoromethylbenzyl chloride (11.6mg, 0.06mmol) were dissolved in DMF (10mL), cesium carbonate (29.3mg, 0.09mmol) was added, and reacted overnight at room temperature to prepare Chromatographic separation gave Example Compound 36 (white solid, 17.1 mg). LC-MS [M+H] ⁺ : m/z 897.5. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.50 (s, 1H), 9.44 (s, 1H), 8.46 (s, 1H), 8.43 (s, 1H), 8.06 (s, 2H), 7.69(s,1H),7.55(s,1H),7.40-7.49(m,6H),6.77(s,1H),6.69(s,1H),5.84-5.90(m,1H),5.50-5.63( m, 1H), 5.21-5.23 (m, 2H), 5.12-5.15 (m, 4H), 4.57-4.59 (m, 2H), 4.48-4.49 (m, 2H), 3.64 (s, 3H), 2.19 ( s, 3H), 2.16 (s, 3H), 1.21 (t, 3H), 1.11 (t, 3H).

Example 37: (E)-9,9'-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-((4-(trifluoromethyl)benzyl)oxo)-9H-pyrimidine[4,5-b]indole-6-carboxamide)

Example compound 32 (30.0mg, 0.04mmol) and p-trifluoromethylbenzyl chloride (23.3mg, 0.12mmol) were dissolved in DMF (10mL), cesium carbonate (39.1mg, 0.12mmol) was added and reacted overnight at room temperature to prepare Chromatographic separation gave Example 37 (white solid, 19.8 mg). LC-MS [M+H] ⁺ : m/z 1041.2. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.48 (s, 2H), 8.51 (s, 2H), 8.09 (s, 2H), 7.67 (s, 2H), 7.47 (s, 2H), 7.34-7.37(m,4H), 7.29-7.31(m,4H), 6.75(s,2H), 5.51(s,2H), 5.14(s,4H), 5.01(s,4H), 4.54-4.56( m, 4H), 2.19 (s, 6H), 1.19 (t, 6H).

Example 38: (E)-9,9'-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-(3-methoxypropoxy)-9H-pyrimidine[4,5-b]indole-6-carboxamide)

Example compound 32 (30.0mg, 0.04mmol) and brominated material (18.2mg, 0.12mmol) were dissolved in DMF (10mL), cesium carbonate (39.1mg, 0.12mmol) was added, and the reaction was carried out at room temperature overnight. Preparative chromatographic separation was carried out. Example 38 (white solid, 20.2mg). LC-MS[M+H] ⁺ : m/z 869.6. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.47(s, 2H), 8.41(s, 2H), 8.05(s, 2H), 7.54(s, 2H), 7.38(s, 2H), 6.75(s, 2H), 5.76(s, 2H), 5.21(s, 4H), 4.53-4.58(m ,4H),3.96-3.99(m,4H),3.21-3.24(m,4H),3.10(s,6H),2.20(s, 6H),1.64-1.68(m,4H),1.20(t,6H) ).

Example 39: (E)-9,9'-(but-2-ene-1,4-diyl)bis(8-(2-(benzyloxy)ethoxy)-2-(1-ethyl 3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-amide)

以2-苄氧基溴乙烷和实施例32为原料，参照实施例38的方法合成得到实施例39(白色固体，12.0mg)。LC-MS[M+H] ⁺:m/z 993.7。 ¹H-NMR(400MHz,DMSO-d ₆):δ9.48(s,2H),8.44(s,2H),8.05(s,2H),7.59(s,2H),7.41(s,2H),7.09-7.17(m,10H),6.75(s,2H),5.65(s,2H),5.10(s,4H),4.52-4.55(m,4H),4.23(s,4H),4.10(s,4H),3.41-3.45(m,4H),2.19(s,6H),1.18(t,6H)。

Using 2-benzyloxy bromoethane and Example 32 as raw materials, referring to the method of Example 38, Example 39 (white solid, 12.0 mg) was synthesized. LC-MS [M+H] ⁺ : m/z 993.7. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.48 (s, 2H), 8.44 (s, 2H), 8.05 (s, 2H), 7.59 (s, 2H), 7.41 (s, 2H), 7.09-7.17 (m, 10H), 6.75 (s, 2H), 5.65 (s, 2H), 5.10 (s, 4H), 4.52-4.55 (m, 4H), 4.23 (s, 4H), 4.10 (s, 4H), 3.41-3.45 (m, 4H), 2.19 (s, 6H), 1.18 (t, 6H).

Example 40: (E)-8-(2-(benzyloxy)ethoxy)-9-(4-(6-acyl-2-(1-ethyl-3-methyl-1H-pyrazole) -5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3- Methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-amide

以实施例34和2-苄氧基溴乙烷为原料，参照实施例38的方法合成得到实施例40(白色固体，13.3mg)。LC-MS[M+H] ⁺:m/z 873.5。 ¹H-NMR(400MHz,DMSO-d ₆):δ9.46(s,2H),8.43(s,2H),8.04-8.07(m,2H),7.58-7.60(m,2H),7.40-7.41(m,2H),7.15-7.21(m,5H),6.72(s,2H),5.70-5.78(m,2H),5.12-5.18(m,4H),4.46-4.58(m,4H),4.30(s,2H),4.17(s,2H),3.74(s,3H),3.56-3.66(m,2H),2.17(d,6H),1.16(t,6H)。

Using Example 34 and 2-benzyloxy bromoethane as raw materials, referring to the method of Example 38, Example 40 (white solid, 13.3 mg) was synthesized. LC-MS [M+H] ⁺ : m/z 873.5. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.46 (s, 2H), 8.43 (s, 2H), 8.04-8.07 (m, 2H), 7.58-7.60 (m, 2H), 7.40-7.41 (m,2H),7.15-7.21(m,5H),6.72(s,2H),5.70-5.78(m,2H),5.12-5.18(m,4H),4.46-4.58(m,4H),4.30 (s, 2H), 4.17 (s, 2H), 3.74 (s, 3H), 3.56-3.66 (m, 2H), 2.17 (d, 6H), 1.16 (t, 6H).

Example 41: (E)-9,9'-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazol-5-yl) )-8-(2-hydroxyethoxy)-9H-pyrimidine[4,5-b]indole-6-amide)

The first step: the compound of Example 39 (10.0 mg, 0.01 mmol) was dissolved in TFA (10 mL), and reacted at 90 degrees overnight. LC-MS detected that the reaction was complete, and the crude product (10.0 mg) was obtained by rotary drying and proceed to the next step. LC-MS [M+H] ⁺ : m/z 1005.5.

Step 2: Dissolve the crude compound (10.0 mg, 0.01 mmol) of the previous step in MeOH/H ₂ O (10 mL/1 mL), add potassium carbonate (4.2 mg, 0.03 mmol), and react at room temperature for 4 hours. LCMS detected that the reaction was complete, and preparative chromatography was separated and purified to obtain Example 41 (white solid, 1.5 mg). LC-MS [M+H] ⁺ : m/z 813.5. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.48 (s, 2H), 8.42 (s, 2H), 8.06 (s, 2H), 7.57 (s, 2H), 7.41 (s, 2H), 6.72 (s, 2H), 5.84 (s, 2H), 5.23-5.25 (m, 4H), 4.83-4.88 (m, 1H), 4.49-4.54 (m, 4H), 3.98-4.01 (m, 4H), 3.37-3.54 (m, 5H), 2.18 (s, 6H), 1.18 (t, 6H).

Example 42: (E)-9-(4-(6-acyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(2-hydroxyethoxy) )-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5- Yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-amide

以实施例40为原料，参照实施例41的方法合成得到实施例42(白色固体，2.5mg)。LC-MS[M+H] ⁺:m/z 783.5。 ¹H-NMR(400MHz,DMSO-d ₆):δ9.46(s,2H),8.41(s,2H),8.05(s,2H),7.59(s,2H),7.41(s,2H),6.72(s,2H),5.84(s,2H),5.18-5.28(m,4H),4.47-4.53(m,4H),4.03-4.05(m,2H),3.78(s,3H),3.50-3.57(m,3H),2.18(s,6H),1.16(t,6H)。

Example 40 was used as a raw material, and Example 42 (white solid, 2.5 mg) was synthesized by referring to the method of Example 41. LC-MS [M+H] ⁺ : m/z 783.5. ¹ H-NMR (400MHz, DMSO-d ₆ ): δ9.46 (s, 2H), 8.41 (s, 2H), 8.05 (s, 2H), 7.59 (s, 2H), 7.41 (s, 2H), 6.72 (s, 2H), 5.84 (s, 2H), 5.18-5.28 (m, 4H), 4.47-4.53 (m, 4H), 4.03-4.05 (m, 2H), 3.78 (s, 3H), 3.50- 3.57 (m, 3H), 2.18 (s, 6H), 1.16 (t, 6H).

Example 43: (E)-9-(4-(6-acyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-8-(3-(diethylamino)propoxy)-2-( 1-Ethyl-3-methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-amide

Example 43 (white solid, 6.1 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 965.6. ¹ H-NMR (400MHz, MeOD-d ₄ ): δ9.28 (s, 2H), 8.27 (s, 1H), 8.26 (s, 1H), 7.48 (s, 1H), 7.39 (s, 1H), 6.84(s, 1H), 6.78(s, 1H), 5.64(s, 2H), 5.16(s, 4H), 4.61-4.66(m, 6H), 3.68-3.73(m, 4H), 3.54-3.57( m,4H),2.36-2.38(m,4H),2.26-2.27(m,6H),2.18-2.22(m,6H),1.47-1.49(m,4H),1.32-1.38(m,6H), 0.92(t,6H).

Example 44: (E)-8-butoxy-9-(4-(6-acyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-( 3-morpholinopropoxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl- 1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-amide

采用实施例25相同的方法制备得到实施例44(白色固体，2.2mg)。LC-MS[M+H] ⁺:m/z 908.6。 ¹H-NMR(400MHz,MeOD-d ₄):δ9.28-9.31(s,2H),8.35(s,1H),8.27(s,1H),7.48(s,1H),7.39(s,1H),6.84(s,1H),6.78(s,1H),5.57-5.71(m,2H),5.05-5.25(m,4H),4.58-4.74(m,4H),3.74-3.95(m,8H),3.14-3.29(m,4H),2.26-2.28(m,6H),1.92-1.96(m,2H),1.22-1.39(m,10H),0.80(t,3H)。

Example 44 (white solid, 2.2 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 908.6. ¹ H-NMR (400MHz, MeOD-d ₄ ): δ9.28-9.31 (s, 2H), 8.35 (s, 1H), 8.27 (s, 1H), 7.48 (s, 1H), 7.39 (s, 1H) ), 6.84 (s, 1H), 6.78 (s, 1H), 5.57-5.71 (m, 2H), 5.05-5.25 (m, 4H), 4.58-4.74 (m, 4H), 3.74-3.95 (m, 8H) ), 3.14-3.29 (m, 4H), 2.26-2.28 (m, 6H), 1.92-1.96 (m, 2H), 1.22-1.39 (m, 10H), 0.80 (t, 3H).

Example 45: (E)-9-(4-(6-acyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholinopropoxy Yl)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-8-(cyclopropylmethoxy)-2-(1-ethyl- 3-Methyl-1H-pyrazol-5-yl)-9H-pyrimidine[4,5-b]indole-6-amide

采用实施例25相同的方法制备得到实施例45(白色固体，12.0mg)。LC-MS[M+H] ⁺:m/z 906.6。 ¹H-NMR(400MHz,MeOD-d ₆):δ9.26(s,1H),8.31(s,1H),8.24(s,1H),7.47(s,1H),7.37(s,1H),6.79(s,1H),6.74(s,1H),5.73-5.79(m,2H),5.25-5.67(m,5H),4.50-4.70(m,3H),3.98-4.02(m,4H),3.65-3.67(m,4H),3.20-3.23(m,4H),2.25-2.29(m,6H),1.98-2.01(m,2H),1.26-1.33(m,6H),0.84-0.95(m,1H),0.42-0.45(m,2H),0.38-040(m,2H)。

Example 45 (white solid, 12.0 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 906.6. ¹ H-NMR (400MHz, MeOD-d ₆ ): δ9.26 (s, 1H), 8.31 (s, 1H), 8.24 (s, 1H), 7.47 (s, 1H), 7.37 (s, 1H), 6.79(s,1H),6.74(s,1H),5.73-5.79(m,2H),5.25-5.67(m,5H),4.50-4.70(m,3H),3.98-4.02(m,4H), 3.65-3.67(m,4H),3.20-3.23(m,4H),2.25-2.29(m,6H),1.98-2.01(m,2H),1.26-1.33(m,6H),0.84-0.95(m , 1H), 0.42-0.45 (m, 2H), 0.38-040 (m, 2H).

Example 46: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxypropyl) Oxy)-9H-pyridyl[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole -5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-amide

采用实施例25相同的方法制备得到实施例46(白色固体，19.1mg)。LC-MS[M+H] ⁺:m/z 909.4. ¹H NMR(400MHz,DMSO-d ₆):δ9.83(s,1H),9.72(s,1H),8.50-8.55(d,1H),8.46(s,1H),8.40(s,1H),8.04-8.08(m,2H),7.61-7.63(d,1H),7.53-7.57(m,2H),7.38-7.46(m,2H),6.76(s,1H),6.61(s,1H),5.66(s,2H),5.18-5.23(m,4H),4.52-4.53(m,2H),4.25-4.29(m,2H),3.98-4.07(m,8H),3.58-3.61(m,2H),3.40-3.43(m,2H),2.94-3.17(m,4H),2.51-2.86(m,2H),2.20(s,6H),1.76-1.78(m,2H),1.60-1.64(m,2H),1.19(t,3H),1.08(t,J＝5.6Hz,3H).

Example 46 (white solid, 19.1 mg) was prepared in the same manner as in Example 25. LC-MS[M+H] ⁺ : m/z 909.4. ¹ H NMR (400MHz, DMSO-d ₆ ): δ 9.83 (s, 1H), 9.72 (s, 1H), 8.50-8.55 (d, 1H ), 8.46 (s, 1H), 8.40 (s, 1H), 8.04-8.08 (m, 2H), 7.61-7.63 (d, 1H), 7.53-7.57 (m, 2H), 7.38-7.46 (m, 2H) ), 6.76 (s, 1H), 6.61 (s, 1H), 5.66 (s, 2H), 5.18-5.23 (m, 4H), 4.52-4.53 (m, 2H), 4.25-4.29 (m, 2H), 3.98-4.07 (m, 8H), 3.58-3.61 (m, 2H), 3.40-3.43 (m, 2H), 2.94-3.17 (m, 4H), 2.51-2.86 (m, 2H), 2.20 (s, 6H) ),1.76-1.78(m,2H),1.60-1.64(m,2H),1.19(t,3H),1.08(t,J=5.6Hz,3H).

Example 47: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxypropyl) Oxy)-9H-pyridyl[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole -5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-amide

采用实施例25相同的方法制备得到实施例47(白色固体，17.5mg)LC-MS[M+H] ⁺:m/z 796.4. ¹H NMR(400MHz,DMSO-d ₆):δ9.47(s,1H),8.51(d,1H),8.42(s,1H),8.37(s,1H),8.01-8.08(m,2H),7.53-7.59(m,3H),7.33-7.42(m,2H),6.73(s,1H),6.58(s,1H),5.68-5.87(m,2H),5.18-5.27(m,4H),4.43-4.48(m,2H),4.24-4.29(m,2H),4.07-4.10(m,2H),3.98(s,3H),3.45-3.48(m,2H),2.18(s,6H),1.64-1.77(m,2H),1.04-1.12(m,6H).

The same method as in Example 25 was used to prepare Example 47 (white solid, 17.5 mg) LC-MS [M+H] ⁺ : m/z 796.4. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.47( s,1H),8.51(d,1H),8.42(s,1H),8.37(s,1H),8.01-8.08(m,2H),7.53-7.59(m,3H),7.33-7.42(m, 2H), 6.73 (s, 1H), 6.58 (s, 1H), 5.68-5.87 (m, 2H), 5.18-5.27 (m, 4H), 4.43-4.48 (m, 2H), 4.24-4.29 (m, 2H),4.07-4.10(m,2H),3.98(s,3H),3.45-3.48(m,2H),2.18(s,6H),1.64-1.77(m,2H),1.04-1.12(m, 6H).

Example 48: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-methoxy Propoxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyridine (Azol-5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例48白色固体(44.1mg).LC-MS[M+H] ⁺:m/z 924.8. ¹H NMR(400MHz,DMSO-d ₆):δ9.50(d,2H),8.47(d,2H),8.44(d,2H),8.08(d,2H),7.56(d,2H),6.76(d,2H),5.62-5.69(m,2H),5.18-5.19(m,4H),4.52-4.61(m,4H),3.87-4.01(m,8H),3.38-3.41(m,4H),3.17-3.22(m,5H),2.89-2.92(m,2H),2.20(d,6H),1.59-1.61(m,2H),1.22-1.25(m,2H),1.19-1.21(m,6H)。

The white solid (44.1mg) of Example 48 was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 924.8. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.50 ( d, 2H), 8.47 (d, 2H), 8.44 (d, 2H), 8.08 (d, 2H), 7.56 (d, 2H), 6.76 (d, 2H), 5.62-5.69 (m, 2H), 5.18 -5.19 (m, 4H), 4.52-4.61 (m, 4H), 3.87-4.01 (m, 8H), 3.38-3.41 (m, 4H), 3.17-3.22 (m, 5H), 2.89-2.92 (m, 2H), 2.20 (d, 6H), 1.59-1.61 (m, 2H), 1.22-1.25 (m, 2H), 1.19-1.21 (m, 6H).

采用实施例25相同的方法制备得到实施例49白色固体(30.1mg).LC-MS[M+H] ⁺:m/z 896.3. ¹H NMR(400MHz,DMSO-d ₆):δ9.48(d,2H),8.45(d,2H),8.08(d,2H),7.58(d,2H),7.44(d,2H),6.73(d,2H),5.62-5.89(m,2H),5.19-5.25(m,4H),4.50-4.55(m,4H),3.89-3.99(m,4H),4.00-4.02(m,2H),3.54-3.61(m,5H),3.28-3.30(m,2H),3.13-3.19(m,2H),2.91-2.94(m,2H),2.17(d,6H),1.15-1.22(m,2H),1.10-1.15(m,6H)。 Example 49: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(2-hydroxyethyl) Oxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole- 5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-carboxamide

The white solid (30.1mg) of Example 49 was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 896.3. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.48 ( d, 2H), 8.45 (d, 2H), 8.08 (d, 2H), 7.58 (d, 2H), 7.44 (d, 2H), 6.73 (d, 2H), 5.62-5.89 (m, 2H), 5.19 -5.25 (m, 4H), 4.50-4.55 (m, 4H), 3.89-3.99 (m, 4H), 4.00-4.02 (m, 2H), 3.54-3.61 (m, 5H), 3.28-3.30 (m, 2H), 3.13-3.19 (m, 2H), 2.91-2.94 (m, 2H), 2.17 (d, 6H), 1.15-1.22 (m, 2H), 1.10-1.15 (m, 6H).

Example 50: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholine) Propoxy)-9H-pyrido[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyridine (Azol-5-yl)-8-methoxy-9H-pyrimid[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例50(白色固体，6.2mg)。LC-MS[M+H] ⁺:m/z 865.2。

Example 50 (white solid, 6.2 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 865.2.

Example 51: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-morpholine) Propoxy)-9H-pyrido[2,3-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyridine (Azol-5-yl)-8-(3-hydroxypropoxy)-9H-pyrimido[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例51(白色固体，5.6mg)。 LC-MS[M+H] ⁺:m/z 909.3。

Example 51 (white solid, 5.6 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 909.3.

Example 52: (E)-9-(4-(8-carbamoyl-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-(3-hydroxypropyl) Oxy)-5H-pyridyl[4,3-b]indol-5-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole -5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例52(白色固体，18.0mg)。LC-MS[M+H] ⁺:m/z 909.5。

Example 52 (white solid, 18.0 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 909.5.

Example 53: (E)-9-(4-(8-carbamoyl-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-(3-hydroxypropyl) Oxy)-5H-pyridyl[4,3-b]indol-5-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole -5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例53(白色固体，18.0mg)。LC-MS[M+H] ⁺:m/z 769.3。

Example 53 (white solid, 18.0 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 769.3.

Example 54: (E)-9-(4-(8-carbamoyl-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-(pyrimidine-2- Methoxy)-5H-pyridyl[4,3-b]indol-5-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H- Pyrazol-5-yl)-8-methoxy-9H-pyrimidine[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例54(白色固体，5.1mg)。LC-MS[M+H] ⁺:m/z 830.2。

Example 54 (white solid, 5.1 mg) was prepared in the same manner as in Example 25. LC-MS [M+H] ⁺ : m/z 830.2.

Example 55: (E)-9-(4-(8-carbamoyl-3-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-6-(3-morpholine) Propoxy)-5H-pyridyl[4,3-b]indol-5-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H- Pyrazol-5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例55淡黄固体(5.0mg).LC-MS[M+H] ⁺:m/z 978.4。 ¹H NMR(400MHz,MeOD-d ₄):δ9.50(s,1H),9.36(s,1H),8.48(s,1H),8.37(s,1H),7.94(s,1H),7.50-7.62(m,2H),6.85(s,1H),6.32(s,1H), 5.72-5.42(m,2H),5.15-5.25(m,4H),4.50-4.71(m,2H),4.17-4.21(m,2H),3.99-4.13(m,8H),3.74-3.81(m,4H),3.40-3.42(m,4H),3.01-3.21(m,8H),2.09-2.28(m,6H),1.85-1.91(m,2H),1.80-1.95(m,2H),1.29-1.30(m,6H)。

The light yellow solid (5.0 mg) of Example 55 was prepared by the same method as that of Example 25. LC-MS [M+H] ⁺ : m/z 978.4. ¹ H NMR (400MHz, MeOD-d ₄ ): δ9.50 (s, 1H), 9.36 (s, 1H), 8.48 (s, 1H), 8.37 (s, 1H), 7.94 (s, 1H), 7.50 -7.62(m,2H),6.85(s,1H),6.32(s,1H), 5.72-5.42(m,2H),5.15-5.25(m,4H),4.50-4.71(m,2H),4.17 -4.21(m,2H),3.99-4.13(m,8H),3.74-3.81(m,4H), 3.40-3.42(m,4H),3.01-3.21(m,8H),2.09-2.28(m, 6H), 1.85-1.91 (m, 2H), 1.80-1.95 (m, 2H), 1.29-1.30 (m, 6H).

Example 56: (E)-9-(4-(6-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazol-5-yl)-8-(3-hydroxypropyl) Oxy)-9H-pyrimidine[4,5-b]indol-9-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole- 5-yl)-8-(3-morpholinopropoxy)-9H-pyrimidinyl[4,5-b]indole-6-carboxamide

采用实施例25相同的方法制备得到实施例56白色固体(9.0mg)。LC-MS[M+H] ⁺:m/z 910.3. ¹H NMR(400MHz,MeOD-d ₄):δ10.13-12.02(m,2H),8.16-8.19(m,2H),7.38-7.41(m,2H),6.61-6.60(m,2H),5.59-5.64(m,2H),5.12-5.33(m,4H),4.44-4.48(m,4H),4.02-4.12(m,2H),3.95-4.01(m,4H),3.93-3.95(m,2H),3.76-3.78(m,2H),3.57-3.58(m,2H),3.30-3.34(m,2H),3.05-3.19(m,2H),1.98-2.20(m,6H),1.73-1.98(m,2H),1.70-1.72(m,2H),1.19-1.28(m,6H)。

The white solid (9.0 mg) of Example 56 was prepared by the same method as that of Example 25. LC-MS[M+H] ⁺ : m/z 910.3. ¹ H NMR (400MHz, MeOD-d ₄ ): δ 10.13-12.02 (m, 2H), 8.16-8.19 (m, 2H), 7.38-7.41 (m,2H),6.61-6.60(m,2H),5.59-5.64(m,2H),5.12-5.33(m,4H),4.44-4.48(m,4H),4.02-4.12(m,2H) ,3.95-4.01(m,4H),3.93-3.95(m,2H),3.76-3.78(m,2H),3.57-3.58(m,2H),3.30-3.34(m,2H),3.05-3.19( m, 2H), 1.98-2.20 (m, 6H), 1.73-1.98 (m, 2H), 1.70 to 1.72 (m, 2H), 1.19-1.28 (m, 6H).

参照专利WO2019069275A第85页实施例4的合成方法制备得到对比化合物1。LC-MS[M+H] ⁺:850.5m/z. ¹H NMR(400MHz,CD ₃OD):δ7.59(s,2H),7.27(s,1H),7.25(s,1H),6.61(s,1H),6.59(s,1H),5.80(br s,2H),5.01(br s,4H),4.57-4.61(m,4H),3.97(br s,4H),3.71(br s,5H),3.12-3.31(m,6H),2.18(s,6H),1.95-1.98(m,2H),1.31-1.39(m,6H)。 Comparative compound 1:

The comparative compound 1 was prepared by referring to the synthesis method of Example 4 on page 85 of patent WO2019069275A. LC-MS[M+H] ⁺ :850.5m/z. ¹ H NMR(400MHz,CD ₃ OD): δ7.59(s,2H),7.27(s,1H),7.25(s,1H),6.61 (s, 1H), 6.59 (s, 1H), 5.80 (br s, 2H), 5.01 (br s, 4H), 4.57-4.61 (m, 4H), 3.97 (br s, 4H), 3.71 (br s) , 5H), 3.12-3.31 (m, 6H), 2.18 (s, 6H), 1.95-1.98 (m, 2H), 1.31-1.39 (m, 6H).

参照专利WO2020028565A1中实施例1的合成方法制备得到对比化合物2。 ¹H NMR(TFA-d,400MHz):δ11.66(s,2H),10.58(d,J＝0.8Hz,2H),9.78(s,2H),9.36(s,2H),8.09(brs,2H),7.51(brs,4H),6.87-6.89(m,4H),5.84(s,6H),4.54(s,6H),3.58(t,J＝7.2Hz,6H)。MS(ESI)：m/z＝753.0[M+H]。 Comparative compound 2:

The comparative compound 2 was prepared by referring to the synthetic method of Example 1 in patent WO2020028565A1. ¹ H NMR (TFA-d, 400MHz): δ11.66(s, 2H), 10.58(d, J = 0.8Hz, 2H), 9.78(s, 2H), 9.36(s, 2H), 8.09(brs, 2H), 7.51 (brs, 4H), 6.87-6.89 (m, 4H), 5.84 (s, 6H), 4.54 (s, 6H), 3.58 (t, J=7.2 Hz, 6H). MS (ESI): m/z=753.0 [M+H].

参照专利WO2020028565A1中实施例2的合成 方法制备得到对比化合物3。LC-MS[M+H] ⁺:m/z 841.4. ¹H NMR(400MHz,DMSO-d ₆):δ9.46(s,2H),8.40(s,2H),8.06(s,2H),7.56(s,2H),7.39(s,2H),6.73(s,2H),5.77(s,4H),5.21(s,4H),4.50-4.54(m,6H),4.01-4.05(m,4H),3.37-3.39(m,4H),2.17(s,6H),1.59-1.63(m,4H),1.14-1.18(m,6H)。 Comparative compound 3:

The comparative compound 3 was prepared by referring to the synthesis method of Example 2 in patent WO2020028565A1. LC-MS[M+H] ⁺ : m/z 841.4. ¹ H NMR (400MHz, DMSO-d ₆ ): δ9.46(s, 2H), 8.40(s, 2H), 8.06(s, 2H), 7.56(s,2H),7.39(s,2H),6.73(s,2H),5.77(s,4H),5.21(s,4H),4.50-4.54(m,6H),4.01-4.05(m, 4H), 3.37-3.39 (m, 4H), 2.17 (s, 6H), 1.59-1.63 (m, 4H), 1.14-1.18 (m, 6H).

Test Example 1: Test for the binding activity of the compound of the example and WT hSTING protein

Follow the test method of the WT hSTING kit provided by Cisbio: In a 96-well screening plate, the STING protein is reacted with the test compound (initial concentration 10μM, ten-fold three-fold dilution) and buffer at 37°C for 30 minutes, then Incubate with 20μM fluorescent substrate at 37°C for 30 minutes. Use a Thermo Scientific Verioskan Flash multi-function reader to use 358nm as excitation light to read the light intensity at 455nm. The measured fluorescence value is compared with the value of the blank well to calculate the competitive binding activity of the sample to the protein. The EC ₅₀ value of the compound was calculated by the Prism software of Graphpad.

The results show that most of the example compounds of the present invention can better bind to WT hSTING protein, and their EC ₅₀ values are less than 100 nM, most of the examples have EC ₅₀ values less than 10 nM, and some of the example compounds have EC 50 values even less than 1 nM. Reach pM level activity. (The specific data is shown in Table 1, A<0.3nM, 0.3nM≤B<1nM, 1nM≤C<10nM, 10nM≤D<100nM, E≥100nM).

Table 1. The binding activity of the example compounds to WT hSTING protein

Numbering EC ₅₀ Numbering EC ₅₀ Numbering EC ₅₀ 1 E 2 E 3 E 4 D 5 C 6 B 7 E 8 E 9 E 10 A 11 A 12 E 13 C 14 E 15 C 16 A 17 C 18 A 19 A 20 B twenty one B twenty two B twenty three B twenty four B 25 C 26 C 27 C 28 C 29 A 30 A 31 B 32 B 33 B 34 C 35 B 36 A 37 A 38 B 39 A 40 A 41 A 42 B 43 C 44 C 45 C 46 A 47 A 48 A 49 A 50 A 51 A 52 A 53 A 54 A 55 A 56 A To To Comparative compound 1 B Comparative compound 2 B Comparative compound 3 C

Test Example 2: Elisa method to detect the release activity test of the compounds of the examples stimulated THP1 cells to IFNβ

Test steps: 1. THP-1 cells are cultured with RPMI1640+10% FBS+0.05mM β-ME, maintaining the density at 2×10 ⁵ ～1×10 ⁶ viable cells/mL; collect the cells, discard the old medium, and use Wash the cells once with serum-free medium, centrifuge to remove the supernatant, and resuspend the cells in serum-free medium. Count after staining with trypan blue. When the cell viability is greater than 95%, perform subsequent experiments; use serum-free medium to adjust the cell density to 1.1×10 ⁶ viable cells/mL; add 180 μL of cell suspension to the wells of the 96-well cell culture plate. The density is 2×10 ⁵ viable cells/well. 2: Dilute the positive control and the test compound with a solvent to form a stock solution and perform a gradient dilution to obtain a 10-fold concentration solution; add 20 μL of a 10-fold concentration compound solution to each well, with two replicate wells for each concentration. The highest concentration of the positive control compound cGAMP is 100 μM, diluted 3 times, and there are 5 concentrations in total. The highest concentration of the compound to be tested is 10μM, 5 times dilution, and a total of 5 concentrations; 3. The cell culture plate is placed in the incubator and incubated for 24hrs; the cell culture plate is centrifuged at 2000g for 5 minutes, and the supernatant is transferred for direct ELISA detection; ELISA test sample The concentration of IFN-β was calculated according to the standard curve of IFN-β. The IFN-β concentration in the cell supernatant is equal to the IFN-β concentration in the ELISA sample multiplied by the dilution factor; 4. Use GraphPad Prism 5.0 software to analyze the data, use the nonlinear S-curve regression to fit the data to obtain the dose-response curve, and calculate the EC ₅₀ value.

Results: Most of the compounds of the examples of the present invention stimulate the release activity of THP1 cells to IFNβ with EC ₅₀ values less than 1 uM, the EC ₅₀ values of some examples are even less than 100 nM, and the EC ₅₀ values of some examples are even less than 1 nM, as in Example 16. , 29, 39, 56 etc. (The specific EC ₅₀ data is shown in Table 2, A<1nM, 1nM≤B<10nM, 10nM≤C<100nM, D≥100nM).

Table 2. Example compounds stimulate the release activity of THP1 cells on IFNβ

Test Example 3: Elisa method to detect the inhibitory ability of the compounds of the examples on the 2'3'-cGAMP-stimulated THP-1 secretion of IFN-β

Test method: 1. THP-1 cells are cultured with RPMI1640+10% FBS+0.05mM β-ME, maintaining the density at 2×10 ⁵ ～1×10 ⁶ viable cells/mL; collect the cells and discard the old medium before use Wash the cells once with serum-free medium, centrifuge to remove the supernatant, and resuspend the cells in serum-free medium. Count after staining with trypan blue, and perform follow-up experiments when the cell viability is greater than 95%. Use serum-free medium to adjust the cell density to 1.1×10 ⁶ viable cells/mL; add 160 μL of cell suspension to the wells of a 96-well cell culture plate, and the cell density is 2×10 ⁵ viable cells/well. 2. Dilute the positive control and the test compound with a solvent to form a stock solution and perform gradient dilution to obtain a 10-fold concentration solution. Add 20 μL of 10-fold concentration compound solution to each well, and incubate in an incubator for 2 hrs. Continue to add 20μL cGAMP to each well, the final concentration of cGAMP is 30μM. The highest concentration of the test compound is 30 μM, diluted 5 times, and there are 5 concentrations in total. Set 1 replicate hole for each concentration. 3. Place the cell culture plate in an incubator for 24hrs, centrifuge at 2000g for 5 minutes, and transfer the supernatant directly for ELISA detection. 4. Refer to the instructions of R&D Systems (Cat#DY814-05) for the ELISA detection process. The concentration of IFN-β in the ELISA test sample was calculated according to the standard curve of IFN-β. The concentration of IFN-β in the cell supernatant is equal to the concentration of IFN-β in the ELISA sample multiplied by the dilution factor. Use GraphPad Prism 5.0 software to analyze the data, use nonlinear S-curve regression to fit the data to obtain a dose-response curve, and calculate the EC ₅₀ value.

Results: The inventors of the present invention in the course of long-term studies, the compound was first discovered Example Compound formula (I) according to the present invention a good antagonist STING portion having functional activity, EC ₅₀ of less than 20 uM, as described in Example 11, 18 , 30, 36, 55; Some of the example compounds have an EC _{50 of} less than 1 uM for 2'3'-cGAMP-stimulated THP-1 secretion of IFN-β, as shown in Examples 11 and 16.

Test Example Four: Inhibitory activity of the compounds of the examples on different enzymes

(1) Preparation of 1×Kinase buffer; (2) Preparation of compound concentration gradient: the test compound test concentration is 10uM starting, 3 times diluted 10 concentrations, and the test is repeated in a 96-well plate. The final concentration is 100 times. 10 different concentrations of solutions. Then use 1×Kinase buffer to further dilute the compounds of each concentration into an intermediate dilution solution of 5 times the final concentration; (3) Add 5 μL of the prepared compound solution to the compound wells of the 384-well plate, and each concentration is tested in a single well. ; Add 5μL of 5% DMSO to negative control wells and positive control wells respectively; (4) Use 1×Kinase buffer to prepare 2.5 times the final concentration of kinase solution; (5) Add 2.5 times of 10μL to compound wells and positive control wells respectively Final concentration of kinase solution; add 10μL of 1×Kinase buffer to the negative control well; (6) Centrifuge at 1000rpm for 30 seconds, shake and mix, and incubate at room temperature for 10 minutes; (7) Use 1×Kinase buffer to prepare 2.5 times the final concentration The mixed solution of ATP and Kinase substrate 22; (8) Add 10 μL of 2.5 times the final concentration of ATP and substrate mixed solution to start the reaction; (9) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake and mix at 28°C, respectively Incubate for the corresponding time; (10) Add 30 μL to stop the detection solution to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, shake and mix; (11) Read the conversion rate with Caliper EZ Reader II, and take the log value of the concentration as the X axis, and the percentage inhibition rate Y-axis, using analysis software of GraphPad Prism log 5 (inhibitor) vs.response-Variable slope fitting dose-response curve to arrive at individual compounds activity IC ₅₀ value pairs. Using the above-mentioned test method, the compounds of the examples were respectively interacted with kinases such as GSK3, TBK, CDK, IKK, AMPK, ULK1, etc., to test the inhibitory activity of the compounds on the above-mentioned enzymes.

Results: Compounds 11, 16, 18, 19, 39, and 56 of the examples of the present invention have weak inhibitory activity against GSK3, TBK, CDK, IKK, AMPK, ULK1 and other kinases, with IC ₅₀ >500 nM, showing a higher binding capacity to STING protein. High selectivity.

All documents mentioned in the present invention are cited as references in this application, as if each document was individually cited as a reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (11)

A nitrogen-containing fused ring compound as shown in general formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, poly Crystal form or prodrug,

Where: R _{1 is} selected from the following group: hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3-6 membered heterocyclic group, 5-10 membered The aryl group, 5-10 membered heteroaryl group, C2-C6 acyl group, C6-C10 sulfonyl group, -S(O) ₂ R ^e , -S(O)R ^e ; wherein the R ₁ group can be Is substituted by one or more substituents selected from R ^c or R ^d , and The R ^{c is} selected from the following group: deuterium, halogen, hydroxyl, amino, 3-8 membered saturated N heterocyclic ring, cyano, nitro, -S(O) ₂ R ^e or -S(O)R ^e , C1-C8 alkyl, 3-15 membered cycloalkyl, 3-15 membered heterocyclic group, C1-C8 alkoxy, C1-C8 alkylamino, alkenyl, alkynyl, C2-C6 acyl or C6-C10 The sulfonyl group, -NHC(O)NHR ^e , or -S(O) ₂ -NHC(O)NHR ^e , C6-C10 sulfinyl group, 5-15 membered aryl group or heteroaryl group; R ^d is selected from the group: deuterium, halo, hydroxy, amino, cyano, nitro, C1-C8 alkyl; or R ^d and R ^a and any one of the atoms are bound, 3- 10-membered ring; R _{2 is} selected from the group consisting of amino, C2-C6 amide, C1-C10 alkyl, 6-10 membered aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, Or a 3-6 membered heterocyclic group; wherein, said R ₂ may be optionally substituted by one or more substituents; _{M 1, M 2, M 3} , M 4, M 5, M 6, M 7 each independently selected from the group consisting of: N, CR ^a; wherein said R ^a is independently selected from the group consisting of: hydrogen, Halogen, hydroxyl, amino, cyano, -C(O)R ^e , -C(O)NHR ^e , -C(O)OR ^e , -NHC(O)NHR ^e , -S(O) ₂ R ^e , -S(O)R ^e , C6-C10 sulfinyl group, C6-C10 sulfinyl group, C6-C10 sulfinimide group, C1-C6 alkyl group, C1-C6 alkoxy group, C2 -C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocyclyl, 5-10 membered aryl or heteroaryl; or any two adjacent R groups above ^a and its connected atoms together form a 4- to 10-membered ring (saturated or partially unsaturated); or M ₃ =M ₄ and M ₅ =M ₆ are each independently selected from O, S or N; Said R ^{e is} selected from the following group: hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkylhydroxyl, C1-C6 alkylalkoxy, C1-C6 alkylamino or substituted amino, C1-C6 alkyl ring amino, C3-C6 cycloalkyl, 3-8 membered heterocyclic group, 6-10 membered aryl group, 5-10 membered heteroaryl group; Wherein, the heteroaryl group contains 1-3 heteroatoms selected from the group: N, O, P or S, and the heterocycloalkyl group contains 1-3 heteroatoms selected from the group: N , O, P or S; Unless otherwise specified, the rings (including the rings in cycloalkyl, heterocyclic, aryl and heteroaryl) include monocyclic, spiro, bridged, fused, and fused rings, and may be saturated or partially unsubstituted. saturation; Unless otherwise specified, each of the above groups can be substituted by one or more (for example, 2, 3, 4, etc.) substituents selected from the following group: halogen, C1-C6 alkoxy, halogenated C1- C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, methyl sulfone, oxo (=O), -CN, hydroxyl, -NH ₂ , C1-C6 amine, carboxyl , C1-C6 amide group (-C(=O)-N(Rf) ₂ or -NH-C(=O)(Rf), Rf is H or C1-C5 alkyl), -OPMB, or substituted or Unsubstituted groups selected from the following group: C1-C6 alkyl, C6-C10 aryl, 5-10 membered heteroaryl with 1-3 heteroatoms selected from N, S and O, -(CH ₂ ) _r -C6-C10 aryl, -(CH ₂ ) _r- (5-10 membered heteroaryl with 1-3 heteroatoms selected from N, S and O),-(having 1-3 5-10 membered heteroarylene group selected from heteroatoms of N, S and O)-(C1-C6 alkyl), 5-12 having 1-3 heteroatoms selected from the group of N, S and O membered heterocyclic group (including monocyclic, spiro, or bridged ring and _{ring), - O (CH 2)} r -3-10 membered cycloalkyl, -O (CH _{2) r} -3-15 membered heterocyclic group , -O(CH ₂ ) _r -C6-C10 aryl, -O(CH ₂ ) _r -5-10-membered heteroaryl; and the substituent is selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 alkylene-OH, C1-C6 alkoxy, oxo, -S(O) ₂ CH ₃ , -CN, -OH, C6-C10 aryl, with 1-3 selected from N, S 3-10 membered heteroaryl group with heteroatom of O, -C(O)CHNH ₂ , -C(O)CHOH; r is selected from 1, 2, 3, 4, 5 or 6; And the compound does not include molecules selected from the following group:

The compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, It is characterized in that the compound of formula (I) is selected from the following group:

_{Wherein, R a1, R a2, R} a3, R a4, R a5, R a6, R a7 each independently selected from the group consisting of: hydrogen, halo, hydroxy, amino, ^{cyano, -C (O) R e,} - ^{C (O) NHR e, -C} (O) OR e, -NHC (O) NHR e ,, - S (O) 2 R e, -S (O) R e, C6-C10 sulfonyl group, C6- C10 sulfinyl group, C6-C10 sulfinimide group, C1-C6 alkyl group, C1-C6 alkoxy group, C2-C6 alkenyl group, C2-C6 alkynyl group, C3-C6 Cycloalkyl, C3-C6 heterocyclyl, 5-10 membered aryl or heteroaryl; or any adjacent of _{R a1} , R _a2 , R _a3 , R _a4 , R _a5 , R _a6 , and R _a7 Both and their connected atoms together form a 4- to 10-membered ring (saturated or partially unsaturated); Or any one of R _a1 , R _a2 , R _a3 , R _a4 , R _a5 , R _a6 , R _a7 and R ^d and the atoms connected between the two together form a 3-10 membered saturated or partially unsaturated ring. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, It is characterized in that the compound of formula (I) is selected from the group

Wherein, R ^2a , R ^2b , and R ^2c are each independently selected from the following group: hydrogen, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl , C3-C6 heterocyclic group; Ra ₄ , Ra ₆ , and R ₁ ′ are each independently selected from the following group: hydrogen, halogen, hydroxyl, amino, cyano, -C(O)R ^e , -C(O)NHR ^e , -C(O)OR ^{e, -NHC (O) NHR e} , -S (O) 2 R e, -S (O) R e, C6-C10 sulfonyl group, C6-C10 C6-C10 sulfinyl group, a sulfinyl alkylene Amino, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocyclic group, 5- 10-membered aryl or heteroaryl; Ma _{8 is} independently selected from NRa ₈ , O, S(O) _p , C(Ra ₈ ) _q , wherein Ra _{8 is} selected from hydrogen, halogen, C1-C6 alkyl, p Is selected from 0-2, q is selected from 1-2; And any one or more hydrogens on the above groups may be substituted by groups selected from the following group: -OR _m , -NR _m R _n , -NR _m COR _n , -CO ₂ R _m , -OCOR _m , -CONR _m R _n , -SO ₂ NR _m R _n , -NR _m SO ₂ R _n , -SR _m , -SOR _m , -SO ₂ R _m , -OCONR _m R _n , -NR _p CONR _m R _n ; R _m , R _n are independently selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkylhydroxyl, C1-C6 alkylalkoxy, C1-C6 alkylamino or substituted amino, C1- C6 alkyl ring amino, C3-C6 cycloalkyl, 3-8 membered heterocyclic group, 6-10 membered aryl group, 5-10 membered heteroaryl group; Or R _m and R _n , R2 _c and R2 _b are each independently connected to the atom to which they are connected, thereby forming a group selected from the following group: 3-12-membered monocyclic or polycyclic alkyl, 3-12-membered monocyclic Cyclic or polycyclic heterocycloalkyl, 3-12 membered spiro or fused ring alkyl, and 3-12 membered spiro or fused ring heterocycloalkyl. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, It is characterized in that the compound of formula (I) has the structure shown in the following formula (II):

Wherein, R _{1 is} selected from the following group: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl; wherein, the R ₁ group may be substituted by one or more R ^d substituents, and R ^d is selected from the group: deuterium, halo, hydroxy, amino, cyano, nitro, C1-C8 alkyl; or R ^d and R ^a and any one of the atoms are bound, 3- 10-membered ring; Ring A is selected from the following group: 9-15 membered heterocyclic group, wherein the heterocyclic group has a condensed polycyclic structure; R ₃ is one or more groups selected from the following group: C1-C6 alkyl, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, 5 -12-membered heterocyclic group; wherein, the heterocyclic group may be substituted by one or more substituents selected from the group consisting of halogen, C1-C6 alkyl; R ₄ is one or more groups selected from the following group: -C(=O)-N(R ^f ) ₂ , -O(CH ₂ ) _r -R ^g , -OPMB; R ^f is H or C1- C5 alkyl; R ^{g is} selected from the following group: H, C6-C10 aryl, 5-10 membered heteroaryl, 3-10 membered cycloalkyl, 3-15 membered heterocyclyl, C1-C6 alkyl, C1-C6 alkoxy, benzyloxy, hydroxyl, C1-C6 amine; and the R ^g may be substituted by one or more halogens. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, It is characterized in that the compound of formula (I) has the structure shown in the following formula (III):

Wherein, the B ring and the C ring are each independently a benzene ring or a 5-7 membered heteroaromatic ring. The compound according to claim 1, 2, 3, which is preferably a compound represented by the following general formula (IV), or a pharmaceutically acceptable salt thereof, or an enantiomer or diastereomer thereof , Tautomers, solvates, polymorphs or prodrugs:

Wherein M ₁ , M ₂ , M ₃ , M ₄ , M ₅ , M ₆ , M ₇ , R _{2 are} as defined in claim 1; R2' is selected from amino, amide, C1-C10 alkyl, 5-10 membered aryl or heteroaryl, C3-C6 cycloalkyl or heterocycloalkyl; M ₁ ', M ₂ ', M ₃ ', M ₄ ', M ₅ ', M ₆ ', M ₇ 'are independently selected from N, CRa, and Ra is independently selected from hydrogen, halogen, hydroxyl, amino, cyano , Carbonyl, amide, ester, urea, sulfone, sulfoxide, sulfonyl, sulfinyl, sulfinimide, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 Alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, 5-10 membered aryl or heteroaryl; or M ₃ '=M ₄ ', M ₅ '=M ₆ 'is independently selected from O, S and other heteroatoms; M is selected from O, NRb, S(O) _t , CRcRd; Rb is independently preferably selected from hydrogen, C1-C10 alkyl, 3-10 membered cycloalkyl or heterocycloalkyl, acyl or sulfonyl; Rc and Rd are independently Ground is selected from hydrogen, deuterium, halogen, C1-C10 alkyl, 3-10 membered cycloalkyl or heterocycloalkyl, or Rc and Rd are connected through a carbon atom or a heteroatom to form a 3-12 membered monocyclic or polycyclic ring Alkyl group, 3-12 membered monocyclic or polycyclic heterocycloalkyl group, 3-12 membered spiro ring or fused ring alkyl group and 3-12 membered spiro ring or fused ring heterocycloalkyl group; m and n are selected from an integer of 0-3; t is selected from an integer of 0-2; The condition is that the structure shown in general formula (IV) does not include the following two molecules:

The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, or polymorph thereof Or a prodrug, characterized in that the compound has the following structure:

The compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof , Characterized in that the compound is selected from the following group:

The compound of formula I according to claims 1-8 or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or The use of the prodrug is characterized in that it is used to prepare drugs for the prevention or treatment of STING protein-dependent diseases. The use according to claim 9, wherein the disease is selected from the following group: tumors, autoimmune diseases, viral infections, and degenerative diseases; preferably, the disease is selected from the following group: non Small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, stomach cancer, bowel cancer, cholangiocarcinoma, brain cancer, leukemia, lymphoma, fibroma, sarcoma, basal cell Cancer, glioma, kidney cancer, melanoma, bone cancer, thyroid cancer, nasopharyngeal cancer, pancreatic cancer, rejection of transplanted organs, gout, rhinitis, alopecia, Alzheimer's disease, appendicitis, atherosclerosis , Asthma, arthritis, allergic dermatitis, Behcet's disease, bullous skin disease, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease, liver cirrhosis, degenerative joint disease, dermatitis, Dermatomyositis, eczema, enteritis, encephalitis, gastritis, nephritis, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease, irritable bowel syndrome, Kawasaki disease, meningitis, multiple sclerosis, myocarditis , Myasthenia gravis, mycosis fungoides, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, Psoriasis, fibrosis, lupus erythematosus, tissue transplant rejection, thyroiditis, type I diabetes, urethritis, uveitis, vasculitis, vitiligo, Waldenstrom's macroglobulinemia, etc. Comprising the compound of formula I according to any one of claims 1-8 or a pharmaceutically acceptable salt thereof, or its enantiomers, diastereomers, tautomers, solvates, A pharmaceutical composition composed of polymorphs or prodrugs, characterized in that the pharmaceutical composition comprises: (i) An effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof ;with (ii) A pharmaceutically acceptable carrier.