DHX9 inhibitor as well as preparation method and application thereof

Abstract

The invention relates to a compound as shown in a formula (1) and a preparation method thereof, a composition containing the compound as shown in the formula (1) and / or pharmaceutically acceptable salt of the composition, a preparation method of the composition and / or the pharmaceutically acceptable salt, and application of the composition as a DHX9 inhibitor in preparation of drugs for treating DHX9-mediated diseases.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a class of DHX9 inhibitors and their preparation methods, as well as the use of such compounds in preparing pharmaceutical compositions for treating, regulating, and / or preventing DHX9-mediated diseases. Background Technology

[0002] DEAH-box helicase 9 (DHX9) is a member of the DexD / H helicase family. It was initially isolated and identified from human and bovine cells. DHX9 is also known as RNA helicase A (RHA) or nuclear DNA helicase II (NDHII). DHX9 possesses NTP-dependent helicase activity. Its active site contains a relatively conserved helicase domain. The N-terminus of the DHX9 protein contains two domains that can bind to double-stranded RNA, and the C-terminus contains a nuclear transport domain and a domain that can bind to single-stranded DNA [Oncotarget, 2016, 7(27): 42716-42739]. DHX9 can promote the unwinding of DNA or RNA complexes and even more complex nucleic acid structures, playing a crucial role in various cellular life activities. DHX9 can regulate DNA replication, RNA transcription, mRNA translation, miRNA generation, RNA processing and transport, and genome stability. Although DHX9 is mainly distributed in the cell nucleus, it can still shuttle into the cytoplasm to regulate the translation of mRNA and the generation of miRNA [J BiolChem.,1995,270(27):16422-16427].

[0003] Abnormal DHX9 expression has a significant impact on cell growth and activity, ultimately leading to disease. DHX9 is highly expressed in various cancers. Two different studies have shown that the expression level of DHX9 in lung tumor tissues is significantly higher than that in normal tissues [Oncol Rep. 2004; 11(1):253-258; BMC Med Genomics. 2014; 7:32], and high DHX9 expression is closely associated with low mortality in patients [BMC Med Genomics. 2014; 7:32]. In breast cancer, the malignancy of tumor tissue is closely related to the expression levels of DHX9, EGFR, and the downstream gene cyclin D1 [ProcNatl Acad Sci USA. 2010, 107(37):16125-16130]. Gene analysis of osteosarcoma cells revealed that the expression level of DHX9 in cells with strong metastatic ability was significantly higher than that in cells with weak metastatic ability. In addition, DHX9, as an NF-κB binding protein, can activate the NF-κB pathway during osteosarcoma metastasis [Int J Oncol.2008,32(1):17-31].

[0004] DHX9 inhibitors are highly dependent on BRCA-deficient tumors (breast cancer, ovarian cancer), mismatch repair deficient (dMMR) or microsatellite instability-high (MSI-H) tumors (colorectal cancer, endometrial cancer, gastric cancer), and hold promise for treating these unmet and in-demand cancers. In addition to cancer, DHX9 is also believed to be associated with viral infections and the development of autoimmune diseases.

[0005] However, there is a lack of DHX9 inhibitors in current clinical practice, and the DHX9 inhibitors in research have shortcomings such as insufficient activity. Therefore, there is an urgent need in this field to develop new DHX9 inhibitors with high activity, high specificity, and / or high safety. Summary of the Invention

[0006] The purpose of this invention is to provide a novel and highly active DHX9 inhibitor, its preparation method, and its application.

[0007] In a first aspect of the invention, a class of compounds of general structural formula as shown in formula (1), or isomers thereof (such as optical isomers), polymorphs, pharmaceutically acceptable salts, hydrates or solvates are provided:

[0008]

[0009] In formula (1):

[0010] m is 0, 1, or 2;

[0011] Ring A is a 5-membered heteroaryl group;

[0012] R 1 Selected from C 1-6 Alkyl or C 3-6 cycloalkyl;

[0013] R 2 Selected from halogen or C 1-3 alkyl;

[0014] R 3 Selected from halogens, CN, C 1-3 Alkyl, C 3-6 cycloalkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, hydroxylated C 1-3 Alkyl, cyano-substituted C 1-3 Alkyl or C 1-3 Alkoxy-substituted C 1-3 alkyl;

[0015] Ring B is selected from: in,

[0016] Indicates a single bond or a double bond;

[0017] n is 0, 1, 2, 3 or 4;

[0018] The ring C is a 4-8 membered heterocyclic alkyl or a 5-8 membered heteroaryl;

[0019] Ring D is C 3-8 Cycloalkylene or 4-8 membered heterocycloalkylene;

[0020] Ring E is C 3-12 Cycloalkylene or 4-12 membered heterocyclic alkylene;

[0021] G 1 and G 2 Each is independently selected from the following groups: chemical bond, O, C(O), CH2, -CH2CH2-, -CH2O- or -OCH2-, S, NR i or * indicates the location linked to ring C, G 1 and G 2 They are not both chemical bonds;

[0022] G 3 and G 4 Each is independently selected from the following groups: chemical bond, O, C(O), CH2, S, or NR. i G 3 and G 4 They are not both chemical bonds;

[0023] G5 Select from the following group: chemical bond, O, C(O) or CH2;

[0024] Each R 5 Independently selected from the following groups: H, halogens, CN, C 1-6 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, cyano-substituted C 1-3 Alkyl or C 1-3 Alkoxy-substituted C 1-3 Alkyl; or, two Rs 5 It forms a -C(O)- with the same carbon atom it is attached to;

[0025] R a R b and R c Independently selected from: H, halogen, C 1-6 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl or C 1-3 Alkoxy-substituted C 1-3 alkyl;

[0026] R d and R e Each is independently selected from the following group: substituted or unsubstituted C 1-6 Alkyl, or substituted or unsubstituted C 3-6 Cycloalkyl; the substitution refers to having one or more (e.g., 2, 3, 4 or 5) substituents selected from the group consisting of: halogen, CN, OH, C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy groups; or R d and R e Together with the attached S atom, it forms a 3-7 member monocyclic heterocyclic alkyl group or a 4-10 member bicyclic heterocyclic alkyl group;

[0027] R f Selected from the following groups: H, halogens, CN, C 1-6 Alkyl or C 1-3 Halogenated alkyl groups;

[0028] R i Selected from the following groups: H, C 1-6 Alkyl or C 1-3 Halogenated alkyl groups.

[0029] In another preferred embodiment, in equation (1), R 1 Selected from the following group: Me, Et,

[0030] In another preferred embodiment, in equation (1), R 2 Select from the following group: Me, Et, F, Cl, or Br.

[0031] In another preferred embodiment, in equation (1), R 3 Selected from the following groups: halogens, CN, Me, Et, CF3, CHF2, OMe, OEt, CH2OH, CH2OMe, CH2CN.

[0032] In another preferred embodiment, in formula (1), ring A is selected from the group consisting of thiophene, pyrrole, pyrazol, furanyl, isothiazolyl or imidazolyl.

[0033] In another preferred embodiment, in equation (1), ring B is selected from the group consisting of:

[0034] Where m, n, ring C, ring D, ring E, and R are... 5 R a R b R c R d R e R f and R i Defined as described above.

[0035] In another preferred embodiment, in equation (1), ring B is selected from the group consisting of:

[0036]

[0037]

[0038]

[0039] In various embodiments, representative compounds of the present invention are selected from the following group:

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047] In a second aspect of the invention, a pharmaceutical composition is provided, the pharmaceutical composition comprising:

[0048] As an active ingredient, a compound, or an isomer thereof, polymorph thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, as described in the first aspect of the present invention; and

[0049] Optionally, a pharmaceutically acceptable excipient or carrier.

[0050] In a third aspect of the invention, the use of the compound, or isomer thereof, polymorph thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, or pharmaceutical composition as described in the first aspect of the invention, in the preparation of a medicament for the prevention and / or treatment of DHX9-mediated related diseases is provided.

[0051] In another preferred example, the DHX9-mediated related diseases are selected from the following group: brain tumors, gastric cancer, liver cancer, lung cancer, intestinal cancer, colon cancer, pancreatic cancer, breast cancer, cervical cancer, endometrial cancer, prostate cancer, leukemia, lymphoma and other solid tumors and hematological malignancies, as well as autoimmune diseases such as rheumatoid arthritis, atopic dermatitis, inflammatory bowel disease, Crohn's disease, allergic rhinitis, lupus erythematosus and other autoimmune diseases.

[0052] In a fourth aspect of the invention, a method for treating, modulating, and / or preventing DHX9-mediated related diseases is provided, comprising: administering to an individual the compound described in the first aspect of the invention, or an isomer thereof, polymorph thereof, pharmaceutically acceptable salt, hydrate, or solvate thereof, or a pharmaceutical composition as described in the second aspect of the invention.

[0053] In another preferred embodiment, the individuals include humans and non-human mammals.

[0054] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0055] Through extensive and in-depth research, and through numerous screenings and tests, the inventors have for the first time discovered a class of compounds of formula (1), which exhibit significant therapeutic effects on DHX9-mediated related diseases. The compounds of this invention also demonstrate good cell proliferation inhibitory activity against the MSI-H HCT-116 cell line, and this invention was completed based on this finding.

[0056] Compounds of the present invention and their synthesis

[0057] This invention provides inhibitors targeting DHX9, namely compounds of formula (1), or isomers thereof, polymorphs thereof, pharmaceutically acceptable salts (inorganic or organic), hydrates, or solvates. Preferably, the compounds of this invention are as described in the first aspect.

[0058] The present invention also provides a method for preparing compounds of formula (1) of the present invention. The method for preparing compounds of formula (1) of the present invention is described in detail below, but these specific methods do not constitute any limitation on the present invention.

[0059] On the one hand, the compounds described herein are prepared according to methods known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to the explanations below. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art. On the other hand, the present invention also provides a method for preparing the compounds of the general formula (1) according to the following general reaction procedures 1-2:

[0060] General reaction procedure 1:

[0061]

[0062] Compounds of general formula (1) can be prepared according to general reaction procedure 1, wherein Y1 and Y1 alone are halogens, boric acids, borate esters or alkyltin groups used for coupling, and R 1 R 2 R 3 Ring A, ring B, and m are as defined above.

[0063] General reaction procedure 2:

[0064]

[0065] Compounds of general formula (1) can be prepared according to general reaction procedure 2, wherein Y1 and Y2 are halogens, boric acids, borate esters or alkyltin groups used for coupling, and R 1 R 2 R 3 Ring A, ring B, and m are as defined above.

[0066] Related definitions

[0067] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0068] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0069] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, trans-butenedioic acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0070] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0071] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures where enantiomers are enriched in diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0072] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0073] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0074] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being a non-mirror image of each other.

[0075] Unless otherwise stated, "(D)" or "(+)" indicates right-handed rotation, "(L)" or "(-)" indicates left-handed rotation, and "(DL)" or "(±)" indicates racemic rotation.

[0076] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0077] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0078] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0079] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 ( 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs generally have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0080] "Optional" or "optionally" means that the event or condition described below may, but is not required, occur, and the description includes both the scenario in which the event or condition occurs and the scenario in which the event or condition does not occur.

[0081] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0082] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and R has independent options in each case. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0083] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a single bond.

[0084] When one of the variables is selected as a chemical bond, it means that the two groups connected are directly linked. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0085] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 It also includes any range from n to n+m, such as C 1-12 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-12 C 6-9 C 6-12 and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-12-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings. They also include any range from n to n+m, such as 3-12-membered rings including 3-6-membered, 3-9-membered, 5-6-membered, 5-7-membered, 6-7-membered, 6-8-membered, and 6-10-membered rings.

[0086] Unless otherwise specified, "C 1-6 "alkyl" is used to denote a straight-chain or branched saturated aliphatic hydrocarbon group consisting of 1 to 6 carbon atoms. The C 1-6 Alkyl groups include C 1-2 C 1-3 C 1-4 C 1-5 C 2-4 C 2-6 C 3-5C1-C6 alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). Non-limiting examples of C1-6 alkyl groups include methyl, ethyl, propyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and their various branched isomers.

[0087] Unless otherwise specified, "C 1-3 "alkyl" is used to denote a straight-chain or branched saturated aliphatic hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Non-limiting examples of alkyl groups include methyl, ethyl, propyl, n-propyl, isopropyl, etc.

[0088] Unless otherwise specified, "C 3-6 "Cycloalkyl" refers to a saturated cyclic aliphatic hydrocarbon group consisting of 3 to 6 carbon atoms, including monocyclic and bicyclic systems. The C... 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C 3-6 Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0089] Unless otherwise specified, "C 1-3 An "alkoxy group" is attached by an oxygen atom to an alkyl group containing 1 to 3 carbon atoms in the remainder of the molecule. The C 1-3 Alkoxy groups include C 1-2 C2 and C3 alkoxy groups, etc.; C 1-3 Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, etc.

[0090] Unless otherwise specified, "heterocyclic alkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or sulfur, wherein the sulfur atom as a ring atom is optionally oxidized, and the nitrogen atom is optionally quaternized. Preferably, it comprises 3 to 12 ring atoms, wherein 1 to 4 are heteroatoms; more preferably, it comprises 3 to 8 ring atoms, wherein 1 to 3 are heteroatoms; most preferably, it comprises 4 to 6 ring atoms, wherein 1 to 3 are heteroatoms. Unless otherwise specified, the ring system of "heterocyclic alkyl" can be a monocyclic, bicyclic, spirocyclic, or polycyclic ring system. "Heterocyclic alkyl" can be attached to the remainder of the molecule via one or more ring carbons or heteroatoms. Non-limiting examples include azirrobutyl, oxacyclobutyl, thioheterobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuranyl-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl, homopiperidinyl, homopiperidinyl, etc.

[0091] Unless otherwise specified, "heteroaryl" refers to a heteroaryl system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, or nitrogen. Heteroaryl groups are preferably 5 to 10-membered, more preferably 5- or 6-membered, such as furanyl, thiophene, pyridinyl, pyrroleyl, N-alkylpyrroleyl, isoxazolyl, thiazolyl, oxazolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc.

[0092] "Cycloalkylene" refers to a saturated cyclic aliphatic hydrocarbon group, which is a residue derived by removing two hydrogen atoms from the same carbon atom of the parent alkane.

[0093] "Heterocyclic alkyl" refers to a saturated cyclic substituent containing one, two, or three heteroatoms selected from N, O, or S, which are residues derived from the removal of two hydrogen atoms from the same carbon atom of the parent alkane.

[0094] "Halogen" or "halogenated" refers to fluorine, chlorine, bromine, or iodine.

[0095] "Hydroxy" refers to -OH.

[0096] “Cyano” refers to -CN.

[0097] Specific pharmaceutical and medical terms

[0098] The term “acceptable,” as used herein, means that a prescription component or active ingredient does not have an excessively harmful effect on health for general therapeutic purposes.

[0099] The terms “treatment,” “treatment process,” or “therapy” as used herein include alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether administered regularly or intermittently, continuously or intermittently, it may be attributable to or related to the administration.

[0100] "Active ingredient" refers to the compound represented by formula (1), and pharmaceutically acceptable inorganic or organic salts of compounds of formula (1). The compounds of the present invention may contain one or more asymmetric centers (axial chirality) and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers. The asymmetric centers that may exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of the present invention. The present invention means including all such isomeric forms of these compounds.

[0101] The terms “compound,” “composition,” “agent,” or “medicine or medicament” may be used interchangeably here, and all refer to a compound or composition that, when applied to an individual (human or animal), can induce a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0102] The term “administered, administering, or administration” here refers to the direct application of the compound or composition described herein, or the application of a prodrug, derivative, or analog of the active compound.

[0103] While the numerical ranges and parameters used to define the broader scope of this invention are approximate values, the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "approximately" may mean that the actual value falls within the acceptable standard error of the mean, as determined by those skilled in the art. Except for experimental examples, or unless explicitly stated otherwise, it is understood that all ranges, quantities, values, and percentages used herein (e.g., to describe material usage, duration, temperature, operating conditions, quantity ratios, and others similar) are modified with "approximately". Therefore, unless otherwise stated, the numerical parameters disclosed in this specification and the accompanying claims are approximate values ​​and are subject to change as needed. At a minimum, these numerical parameters should be understood as the indicated significant digits and values ​​obtained using general rounding.

[0104] Unless otherwise defined in this specification, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, unless conflicting with the context, singular nouns used herein include their plural forms, and vice versa.

[0105] route of administration

[0106] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into various preparations, comprising, within a safe and effective range, the compounds of this invention or their pharmaceutically acceptable salts and pharmacologically acceptable excipients or carriers. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective range of the compound is determined based on the age, condition, and duration of treatment of the patient.

[0107] "Pharmaceutically acceptable excipients or carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0108] When applying the compounds of this invention, they can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), or topically.

[0109] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with: (a) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0110] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0111] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0112] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0113] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0114] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0115] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0116] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0117] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to the mammal (such as a human) requiring treatment. The dosage at the time of administration is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 50–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0118] The main advantages of this invention include:

[0119] (a) Surprisingly, the compound of formula (1) in this invention exhibits excellent inhibitory activity against DHX9 enzyme;

[0120] (b) The compounds of the present invention have stronger antiproliferative activity against HCT116 cells;

[0121] (c) The compounds of the present invention have good drug-like properties.

[0122] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0123] The following description will elaborate on the specific aspects, characteristics, and advantages of the above-described compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples below depict specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0124] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the grown single crystals are used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα phase emission as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0125] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed as δ (ppm). Unless otherwise specified, the silica gel used for separation was 200-300 mesh, and all eluent ratios were by volume.

[0126] The following abbreviations are used in this invention: AcOH represents acetic acid; ACN(CH3CN) represents acetonitrile; Ag2O represents silver oxide; AIBN represents azobisisobutyronitrile; BF3·Et2O represents boron trifluoride diethyl ether; CAN represents cerium ammonium nitrate; Cs2CO3 represents cesium carbonate; CuI represents cuprous iodide; EA(EtOAc) represents ethyl acetate; DCM represents dichloromethane; DEAD represents diethyl azobiscarboxylate; DIBAL-H represents diisobutylaluminum hydride; DIPEA(DIEA) represents diisopropylethylamine; Dioxane represents 1,4-dioxane; DMAP represents 4-dimethylaminopyridine; DMF represents N,N-dioxane. Methylformamide; DMSO represents dimethyl sulfoxide; DPEPHOS represents bis(2-diphenylphosphine) ether; EtOH represents ethanol; Fe represents iron; h represents hours; HATU represents 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphonate; HMDSNa represents sodium bis(trimethylsilylamino)amine; H3PO2 represents hypophosphite; IPA represents isopropanol; K2CO3 represents potassium carbonate; KI represents potassium iodide; K3PO4 represents potassium phosphate. LiAlH4 represents lithium aluminum hydride; MeOH represents methanol; MS represents mass spectrometry; MTBE represents methyl tert-butyl ether; N2 represents nitrogen; NaBH4 represents sodium borohydride; NaBH3CN represents sodium cyanoborohydride; NaH represents sodium hydride; NaNO2 represents sodium nitrite; NaOH represents sodium hydroxide; NBS represents N-bromosuccinimide; NH4Cl represents ammonium chloride; NMI represents N-methylimidazolium; NMP represents N-methylpyrrolidone; NMR represents... Nuclear magnetic resonance; Pd2(dba)3 represents tris(dibenzylacetone)palladium; PE represents petroleum ether; Pd(dppf)Cl2 represents 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; PhI(OAc)2 represents iodophenyldiacetic acid; Ph3P represents triphenylphosphine; PTS represents p-toluenesulfonic acid; SOCl2 represents thionyl chloride; TBAB represents tetra-n-butylammonium bromide; TBAF represents tetra-n-butylammonium fluoride; TBSCl represents tert-butyldimethylchlorosilane; t- BuONa represents sodium tert-butoxide; TCFH represents tetramethylchlorourea hexafluorophosphate; TFA represents trifluoroacetic acid; Tf2O represents trifluoroformic anhydride; TLC represents thin-layer chromatography; THF represents tetrahydrofuran; Xantphos represents 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene; XPhos represents 2-dicyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl.

[0127] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0128] Preparation Example 1: Synthesis of Intermediate 1

[0129]

[0130] Step 1: Synthesis of N-(3-chloro-5-nitrophenyl)methanesulfonamide

[0131] 1-Chloro-3-fluoro-5-nitrobenzene (10 g, 56.97 mmol) and methanesulfonamide (5.42 g, 56.97 mmol) were dissolved in DMSO (80 mL), and Cs₂CO₃ (55.7 g, 170.91 mmol) was added. The mixture was reacted under argon protection at 60 °C with stirring for 2 h. LC-MS monitoring showed that the reaction was complete. The solution was poured into ice water, extracted with EA, concentrated, and subjected to column chromatography (PE / EA = 30 / 1 to 5 / 1) to give a pale yellow solid (8.57 g, yield 60%), ESI-MS m / z: 250.0 [M+H]. + .

[0132] Step 2: Synthesis of N-(3-amino-5-chlorophenyl)methanesulfonamide

[0133] N-(3-chloro-5-nitrophenyl)methanesulfonamide (8.57 g, 34.2 mmol) was dissolved in EtOH / H₂O (5 / 1, 150 mL), and iron powder (9.55 g, 171 mmol) and NH₄Cl (9.15 g, 171 mmol) were added. The mixture was refluxed at 80 °C for 3 h under nitrogen protection. LC-MS monitoring indicated that the reaction was complete. The mixture was filtered, and the filtrate was diluted with water (100 mL). Extraction was performed using EA (100 mL x 3). The organic phases were combined, dried, filtered, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1). The concentrated product yielded a pale yellow solid (5.66 g, 75% yield). ESI-MS m / z: 221.0 [M+H] + .

[0134] Step 3: Synthesis of 4-bromo-N-(3-chloro-5-(methylsulfonamido)phenyl)thiophene-2-carboxamide

[0135] 4-Bromothiophene-2-carboxylic acid (4.14 g, 20 mmol) was dissolved in ACN (100 mL), and N-(3-amino-5-chlorophenyl)methanesulfonamide (4.4 g, 20 mmol), NMI (5.75 g, 70 mmol), and TCFH (8.42 g, 30 mmol) were added. The reaction was carried out at room temperature for 5 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. The solution was diluted with EA, washed with water, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1). The concentrated solution yielded a pale yellow solid (6.55 g, 80% yield). ESI-MS m / z: 408.9 [M+H] + .

[0136] Step 4: Synthesis of (5-((3-chloro-5-(methylsulfonamido)phenyl)carbamoyl)thiophene-3-yl)boronic acid

[0137] 4-Bromo-N-(3-chloro-5-(methylsulfonamido)phenyl)thiophene-2-carboxamide (6.55 g, 16.0 mmol) was dissolved in Dioxane (150 mL), followed by 5,5,5',5'-tetramethyl-2,2'-bis(1,3,2-dioxaborane) (3.61 g, 16.0 mmol), Pd(dppf)Cl2 (585 mg, 0.8 mmol), and K3PO4 (6.8 g, 32.0 mmol). The mixture was stirred at 100 °C for 8 h, and the reaction was monitored by LC-MS until completion. The solution was poured into water (100 mL), extracted with EA (50 mL * 3), and the organic phases were combined, dried, concentrated, and subjected to reversed-phase column chromatography. The resulting solution was lyophilized to give a pale yellow solid (3.75 g, yield 63%). ESI-MS m / z: 375.0 [M+H] + .

[0138] Example 1: N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3-((1-tetrahydro-1λ) 6 Synthesis of thiophene-1-yl)amino)pyridin-2-yl)thiophene-2-carboxamide (compound 1)

[0139]

[0140] Step 1: 1-Iminotetrahydro-1H-λ 6 Synthesis of 1-thiophene oxide

[0141] Tetrahydrothiophene (500 mg, 5.68 mmol) was dissolved in MeOH (30 mL), and iodophenylacetic acid (3.66 g, 11.36 mmol) and ammonium carbonate (1.09 g, 11.36 mmol) were added. The mixture was stirred at room temperature, and the reaction was monitored by LC-MS until the starting material was completely reacted. The solution was concentrated and subjected to column chromatography (DCM / MeOH = 30 / 1) to give a colorless oil (500 mg, yield 74%), ESI-MS m / z: 120.0 [M+H). + .

[0142] Step 2: 1-((2-chloropyridin-3-yl)imino)tetrahydro-1H-λ 6 Synthesis of 1-thiophene oxide

[0143] 1-Imintetrahydro-1H-λ 6 Thiophene-1-oxide (500 mg, 4.20 mmol) and 3-bromo-2-chloropyridine (806 mg, 4.20 mmol) were dissolved in Dioxane (30 mL). Pd₂(dba)₃ (384 mg, 0.42 mmol), Xantphos (243 mg, 0.42 mmol), and CS₂CO₃ (2.74 g, 8.40 mmol) were added. The mixture was stirred at 110 °C and monitored by LC-MS. After the reaction was complete, water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (DCM / MeOH = 30 / 1) to give a yellow solid (430 mg, yield 44%). ESI-MS m / z: 231.0 [M + H₂]. + .

[0144] Step 3: N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3-((1-tetrahydro-1λ) 6 -thiophene-1-yl)amino)pyridin-2-yl)thiophene-2-carboxamide

[0145] 1-((2-chloropyridin-3-yl)imino)tetrahydro-1H-λ6-thiophene-1-oxide (116 mg, 0.5 mmol) and (5-((3-chloro-5-(methylsulfonamido)phenyl)carbamoyl)thiophene-3-yl)boronic acid (187 mg, 0.5 mmol) were dissolved in Dioxane / H2O (10 / 1, 10 mL), and Pd(dppf)Cl2 (37 mg, 0.05 mmol) and K3PO4 (212 mg, 1.0 mmol) were added. The mixture was stirred at 80 °C for 1 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. The solution was poured into water (20 mL), extracted with EA (20 mL * 3), the organic phases were combined, dried, concentrated, and subjected to reversed-phase column chromatography to obtain an off-white solid (107 mg, yield 41%).

[0146] 1 H NMR (400MHz, DMSO-d6) δ: 10.64 (s, 1H), 10.09 (s, 1H), 8.86 (d, J = 20.3Hz, 2H), 8.23(d,J=4.0Hz,1H),7.73(s,2H),7.52(d,J=7.6Hz,1H),7.23(dd,J=8.1,4.5 Hz,1H),6.96(s,1H),3.63-3.53(m,2H),3.40(dd,J=13.1,6.9Hz,2H),3.10(s ,3H),2.26(dd,J=12.8,6.2Hz,2H),2.20-2.11(m,2H); ESI-MSm / z:525.0[M+H] + .

[0147] Example 2-11: Synthesis of compound 2-11

[0148] Compound 2-11 can be synthesized from the raw materials shown in the table according to a method similar to that in Example 1.

[0149]

[0150]

[0151] Example 12: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(5,6,7,8-tetrahydrobenzofurano[2,3-c]pyridin-1-yl)thiophene-2-carboxamide (compound 12)

[0152]

[0153] Step 1: Synthesis of 2-chloro-3-(methoxymethoxy)pyridine

[0154] 2-Chloropyridine-3-ol (10 g, 76.9 mmol) and Na₂CO₃ (16.3 g, 153.8 mmol) were dissolved in ACN (100 mL), and bromo(methoxy)methane (10.6 g, 84.6 mmol) was added dropwise at 0 °C. The reaction was carried out at room temperature for 1 h. LC-MS monitoring showed that the reaction was complete. The reaction solution was filtered, the filtrate was concentrated, and column chromatography (PE / EA = 50 / 1 to 5 / 1) was performed. The concentrate was then concentrated to give a white solid (10 g, yield 74%), ESI-MS m / z: 174.0 [M+H]. + .

[0155] Step 2: Synthesis of 2-chloro-4-iodo-3-(methoxymethoxy)pyridine

[0156] 2-Chloro-3-(methoxymethoxy)pyridine (5.8 g, 33.4 mmol) was added to a 250 mL three-necked flask and dissolved in anhydrous THF (80 mL). Under argon protection, the mixture was cooled to -78 °C, and n-BuLi (2.5 M inhexane, 33.4 mL, 83.5 mmol) was slowly added dropwise. The mixture was kept at this temperature and stirred for 1 h. Then, iodine (8.5 g, 33.4 mmol) in THF solution (20 mL) was added dropwise, and the mixture was kept at this temperature and stirred for 1 h. The reaction was monitored by LC-MS until complete. The reaction was quenched with NH4Cl aqueous solution, extracted with EA, dried, filtered, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1). The concentrated solid yielded a pale yellow solid (7.2 g, 72% yield). ESI-MS m / z: 299.9 [M+H] + .

[0157] Step 3: Synthesis of 2-(2-chloro-3-(methoxymethoxy)pyridin-4-yl)cyclohexane-1-one

[0158] 2-Chloro-4-iodo-3-(methoxymethoxy)pyridine (1.2 g, 4.0 mmol) was dissolved in Dioxane (30 mL), and cyclohexanone (785 mg, 8.0 mmol), XantPhos (231 mg, 0.4 mmol), Pd2(dba)3 (366 mg, 0.4 mmol), and Cs2CO3 (3.9 g, 12.0 mmol) were added. The mixture was purged with argon and heated to 100 °C overnight. The reaction was monitored by LC-MS until complete. The mixture was filtered, directly concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a yellow solid (540 mg, 50% yield). ESI-MS m / z: 270.0 [M+H] + .

[0159] Step 4: Synthesis of 1-chloro-5,6,7,8-tetrahydrobenzofurano[2,3-c]pyridine

[0160] 2-(2-chloro-3-(methoxymethoxy)pyridin-4-yl)cyclohexane-1-one (540 mg, 2.0 mmol) was dissolved in DCM (15 mL), and TFA (2 mL) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by LC-MS until complete. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1), yielding a pale yellow solid (253 mg, 61% yield). ESI-MS m / z: 208.1 [M+H] + .

[0161] Step 5: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(5,6,7,8-tetrahydrobenzofurano[2,3-c]pyridin-1-yl)thiophene-2-carboxamide

[0162] Compound 12 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0163] 1 H NMR (400MHz, DMSO-d6) δ: 10.64 (s, 1H), 9.06 (s, 1H), 8.89 (s, 1H), 8.40 (d, J = 5.4Hz, 1H), 7.74 (s, 2H), 7.57 (d,J=5.4Hz,1H),6.98(s,1H),3.08(s,3H),2.63-2.53(m,4H),1.90-1.79(m,4H); ESI-MSm / z:502.1[M+H] + .

[0164] Examples 13-14: Synthesis of compounds 13-14

[0165] Compounds 13-14 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 12.

[0166] Example 15: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3,4-dihydro-1H-pyrano[4',3':4,5]thieno[2,3-c]pyridin-8-yl)thiophene-2-carboxamide (compound 15)

[0167]

[0168] Step 1: Synthesis of 4-(3-bromo-2-chloropyridin-4-yl)dihydro-2H-pyran-3(4H)-one

[0169] 2-Chloro-3-bromo-4-iodopyridine (1.27 g, 4.0 mmol, synthesis reference ACS Catalysis, 2023, vol. 13, #6, pp. 3788-3793) was dissolved in toluene (50 mL), and dihydro-2H-pyran-3(4H)-one (800 mg, 8.0 mmol), XantPhos (231 mg, 0.4 mmol), Pd2(dba)3 (366 mg, 0.4 mmol), and Cs2CO3 (3.9 g, 12.0 mmol) were added. The mixture was purged with argon, and the temperature was raised to 100 °C and reacted overnight. The reaction was monitored by LC-MS until complete. The mixture was filtered, directly concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a yellow solid (465 mg, yield 40%). ESI-MS m / z: 290.0 [M+H] + .

[0170] Step 2: Synthesis of 4-(3-bromo-2-chloropyridin-4-yl)dihydro-2H-pyran-3(4H)-thione

[0171] 4-(3-bromo-2-chloropyridin-4-yl)dihydro-2H-pyran-3(4H)-one (465 mg, 1.6 mmol) and phosphorus pentasulfide (77 mg, 0.4 mmol) were suspended in toluene (3 mL) under argon protection and stirred at room temperature for 10 min. Hexamethyldisiloxane was added, and the mixture was heated to 90 °C and reacted overnight. The reaction was monitored by LC-MS until completion. The mixture was filtered and directly concentrated to give a pale yellow solid (295 mg, 60% yield). ESI-MS m / z: 305.9 [M+H] + It is used directly in the next reaction.

[0172] Step 3: Synthesis of 8-chloro-3,4-dihydro-1H-pyrano[4',3':4,5]thiopheno[2,3-c]pyridine

[0173] DPEPos (54 mg, 0.1 mmol), Pd2(dba)3 (92 mg, 0.1 mmol), and Cs2CO3 (489 mg, 1.5 mmol) were suspended in toluene (5 mL), purged with argon, and 4-(3-bromo-2-chloropyridin-4-yl)dihydro-2H-pyran-3(4H)-thione (295 mg, 0.96 mmol) was added. The mixture was heated to 100 °C and reacted overnight. The reaction was monitored by LC-MS until complete. The mixture was filtered, concentrated directly, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a yellow solid (91 mg, yield 42%), ESI-MS m / z: 226.0 [M+H]. + .

[0174] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3,4-dihydro-1H-pyrano[4',3':4,5]thieno[2,3-c]pyridin-8-yl)thiophene-2-carboxamide

[0175] Compound 15 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0176] 1H NMR(400MHz, DMSO-d6)δ:10.72(s,1H),9.16(s,1H),9.04(s,1H),8.40(d,J=5.4Hz,1H),7.74(s,2H),7.57(d,J=5.4 Hz,1H),6.98(s,1H),4.95(s,2H),3.85(t,J=5.3Hz,2H),3.65(d,J=5.4Hz,2H),3.10(s,3H); ESI-MSm / z:520.0[M+H] + .

[0177] Example 16: Synthesis of Compound 16

[0178] Compound 16 can be synthesized from the starting materials shown in the table using a similar method to that used for Compound 15.

[0179]

[0180] Example 17: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(1,3,4,9-tetrahydropyrano[4',3':4,5]pyrrolo[2,3-c]pyridin-8-yl)thiophene-2-carboxamide (compound 17)

[0181]

[0182] Step 1: 4-(2-chloro-3-nitropyridin-4-yl)dihydro-2H-pyran-3(4H)-one

[0183] 2-Chloro-3-nitro-4-iodopyridine (1.14 g, 4.0 mmol, synthesis reference: Chemical Science, 2020, vol. 11, #7, pp. 1855-1861) was dissolved in toluene (50 mL). Dihydro-2H-pyran-3(4H)-one (800 mg, 8.0 mmol), XantPhos (231 mg, 0.4 mmol), Pd2(dba)3 (366 mg, 0.4 mmol), and Cs2CO3 (3.9 g, 12.0 mmol) were added. The mixture was purged with argon and heated to 100 °C overnight. The reaction was monitored by LC-MS until complete. The mixture was filtered, concentrated directly, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a yellow solid (257 mg, yield 25%). ESI-MS m / z: 257.0 [M+H] + .

[0184] Step 2: 8-Chloro-1,3,4-9-tetrahydropyrano[4',3':4,5]pyrrolo[2,3-c]pyridine

[0185] 4-(2-chloro-3-nitropyridin-4-yl)dihydro-2H-pyran-3(4H)-one (257 mg, 1.0 mmol) was dissolved in EtOH / H2O (5 / 1, 20 mL), and iron powder (196 mg, 3.5 mmol), acetic acid (424 mg, 7.0 mmol), and sodium acetate dihydrate (118 mg, 1.0 mmol) were added. The mixture was refluxed under argon protection for 2 h. The reaction was monitored by LC-MS until completion. The mixture was filtered, concentrated, and the residue was dissolved in EA, washed with water, and the organic phase was concentrated. Column chromatography (PE / EA = 50 / 1 to 1 / 1) was performed to give a yellow solid (100 mg, yield 48%), ESI-MS m / z: 209.0 [M+H]. + .

[0186] Step 3: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(1,3,4,9-tetrahydropyrano[4',3':4,5]pyrrolo[2,3-c]pyridin-8-yl)thiophene-2-carboxamide

[0187] Compound 17 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0188] 1 H NMR(400MHz, DMSO-d6)δ:11.04(s,1H),10.62(s,1H),9.02(s,1H),8.96(s,1H),8.36(d,J=5.4Hz,1H),7.74(s,2H),7.57(d ,J=5.4Hz,1H),6.98(s,1H),4.90(s,2H),3.75(t,J=5.3Hz,2H),3.49(d,J=5.4Hz,2H),3.08(s,3H); ESI-MSm / z:503.1[M+H] + .

[0189] Example 18: Synthesis of Compound 18

[0190] Compound 18 can be synthesized from the starting materials shown in the table using a similar method to that used for Compound 17.

[0191]

[0192] Example 19: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3,4-dihydro-1H-pyrido[3',4':4,5]imidazo[2,1-c][1,4]oxazin-9-yl)thiophene-2-carboxamide (compound 19)

[0193]

[0194] Step 1: Synthesis of N-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-chloro-3-nitropyridine-4-amine

[0195] 2,4-Dichloro-3-nitropyridine (1.92 g, 10 mmol) was dissolved in DMF (15 mL), and 2-((tert-butyldimethylsilyl)oxy)ethyl-1-amine (1.93 g, 11.0 mmol) and TEA (2.02 g, 20 mmol) were added. The mixture was reacted at 50 °C for 5 h under argon protection, and the reaction was monitored by LC-MS until completion. The solution was diluted with EA, washed with water, and the organic phase was concentrated. Column chromatography (PE / EA = 50 / 1 to 2 / 1) was performed to give a yellow solid (2.1 g, yield 63%). ESI-MS m / z: 332.1 [M+H] + .

[0196] Step 2: N 4 Synthesis of 2-((tert-butyldimethylsilyl)oxy)ethyl)-2-chloropyridine-3,4-diamine

[0197] N-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-chloro-3-nitropyridine-4-amine (2.1 g, 6.3 mmol) was dissolved in EtOH / H₂O (5 / 1, 100 mL), and iron powder (2.1 g, 37.8 mmol) and NH₄Cl (2.02 g, 37.8 mmol) were added. The mixture was refluxed at 80 °C for 1 h under nitrogen protection. LC-MS monitoring indicated that the reaction was complete. The mixture was filtered, and the filtrate was diluted with water (30 mL). Extraction was performed using EA (50 mL x 3). The organic phases were combined, dried, filtered, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1). The concentrated product yielded a pale yellow solid (1.8 g, 95% yield). ESI-MS m / z: 302.1 [M+H] + .

[0198] Step 3: Synthesis of 1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridine

[0199] N 41,8-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-chloropyridine-3,4-diamine (1.8 g, 6.0 mmol) was dissolved in ACN (50 mL), and 2-chloro-1,1,1-trimethoxyethane (1.02 g, 6.6 mmol) and TsOH·H₂O (114 mg, 0.6 mmol) were added. The mixture was heated to 60 °C and reacted for 3 h. LC-MS monitoring showed that the reaction was complete. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1), yielding a pale yellow solid (1.51 g, 70% yield). ESI-MS m / z: 360.1 [M+H] + .

[0200] Step 4: Synthesis of 2-(4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridin-1-yl)ethyl-1-ol

[0201] 1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridine (1.51 g, 4.2 mmol) was dissolved in THF (30 mL), and a THF solution of TBAF (4.2 mL, 1.0 M in THF) was added. The reaction was carried out at room temperature for 1 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1), and the concentrate was concentrated to give a pale yellow solid (827 mg, yield 80%), ESI-MS m / z: 246.1 [M+H]. + .

[0202] Step 5: Synthesis of 9-chloro-3,4-dihydro-1H-pyrido[3',4':4,5]imidazo[2,1-c][1,4]oxazine

[0203] 2-(4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridin-1-yl)ethyl-1-ol (820 mg, 3.2 mmol) was dissolved in THF (20 mL), placed in an ice bath, and NaH (141 mg, 3.52 mmol, 60% in oil) was added. The reaction was carried out at room temperature for 3 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. The reaction was quenched with water, extracted with EA, and the organic phase was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1). The concentrated product yielded a pale yellow solid (420 mg, yield 63%). ESI-MS m / z: 210.0 [M+H] + .

[0204] Step 6: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3,4-dihydro-1H-pyrido[3',4':4,5]imidazo[2,1-c][1,4]oxazin-9-yl)thiophene-2-carboxamide

[0205] Compound 19 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0206] 1 H NMR (400MHz, DMSO-d6) δ: 10.70 (s, 1H), 9.13 (s, 1H), 9.04 (s, 1H), 8.44 (d, J = 5.4Hz, 1H), 7.71 (d, J = 1.7Hz, 2H), 7.62 (d, J =5.4Hz,1H),6.96(s,1H),5.11(s,2H),4.31(d,J=5.0Hz,2H),4.24(d,J=5.2Hz,2H),3.07(s,3H); ESI-MSm / z:504.1[M+H] + .

[0207] Examples 20-21: Synthesis of compounds 20-21

[0208] Compounds 20-21 can be synthesized from the starting materials shown in the table in a manner similar to that in Example 19.

[0209]

[0210]

[0211] Example 22: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6,7,8,9-tetrahydropyridino[3',4':4,5]imidazo[1,2-a]pyrazin-1-yl)thiophene-2-carboxamide (compound 22)

[0212]

[0213] Step 1: Synthesis of tert-butyl (2-((2-chloro-3-nitropyridin-4-yl)amino)ethyl)carbamate

[0214] 2,4-Dichloro-3-nitropyridine (1.92 g, 10 mmol) was dissolved in DMF (15 mL), and tert-butyl (2-aminoethyl)carbamate (1.76 g, 11.0 mmol) and TEA (2.02 g, 20 mmol) were added. The mixture was reacted at 50 °C for 5 h under argon protection, and the reaction was monitored by LC-MS until completion. The solution was diluted with EA, washed with water, and the organic phase was concentrated. Column chromatography (PE / EA = 50 / 1 to 2 / 1) was performed to give a yellow solid (2.5 g, yield 79%), ESI-MS m / z: 317.1 [M+H]. + .

[0215] Step 2: Synthesis of tert-butyl (2-((3-amino-2-chloropyridin-4-yl)amino)ethyl)carbamate

[0216] 2.5 g (7.9 mmol) of tert-butyl (2-((2-chloro-3-nitropyridin-4-yl)amino)ethyl)carbamate was dissolved in EtOH / H₂O (5 / 1, 100 mL), and iron powder (2.65 g, 47.4 mmol) and NH₄Cl (2.54 g, 47.4 mmol) were added. The mixture was refluxed at 80 °C for 1 h under nitrogen protection. LC-MS monitoring indicated that the reaction was complete. The mixture was filtered, and the filtrate was diluted with water (50 mL). Extraction was performed using EA (50 mL x 3). The organic phases were combined, dried, filtered, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1). The concentrate yielded a pale yellow solid (2.0 g, 88% yield). ESI-MS m / z: 287.1 [M+H] + .

[0217] Step 3: Synthesis of tert-butyl (2-(4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridin-1-yl)ethyl)carbamate

[0218] 2-((3-amino-2-chloropyridin-4-yl)amino)ethyl)tert-butyl carbamate (2.0 g, 7.0 mmol) was dissolved in ACN (50 mL), and 2-chloro-1,1,1-trimethoxyethane (1.19 g, 7.7 mmol) and TsOH·H₂O (133 mg, 0.7 mmol) were added. The mixture was heated to 60 °C and reacted for 3 h. LC-MS monitoring showed that the reaction was complete. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1). The concentrate yielded a pale yellow solid (1.5 g, yield 62%). ESI-MS m / z: 345.1 [M+H] + .

[0219] Step 4: Synthesis of tert-butyl 1-chloro-6,7-dihydropyrido[3',4':4,5]imidazo[1,2-a]pyrazine-8(9H)-carboxylic acid

[0220] 1.5 g (4.3 mmol) of tert-butyl 2-(4-chloro-2-(chloromethyl)-1H-imidazo[4,5-c]pyridin-1-yl)ethyl)carbamate was dissolved in 30 mL of THF. The mixture was placed in an ice bath, and NaH (190 mg, 4.77 mmol, 60% in oil) was added. The reaction was carried out at room temperature for 1 h, then heated to 50 °C for 3 h. LC-MS monitoring indicated the reaction was complete. The reaction was quenched with water, extracted with EA, and the organic phase was directly concentrated. Column chromatography (PE / EA = 50 / 1 to 2 / 1) was performed, and the concentrated product yielded a pale yellow solid (1.1 g, 83% yield). ESI-MS m / z: 309.1 [M+H]+ .

[0221] Step 5: Synthesis of 1-chloro-6,7,8,9-tetrahydropyrido[3',4':4,5]imidazo[1,2-a]pyrazine

[0222] 1-Chloro-6,7-dihydropyrido[3',4':4,5]imidazo[1,2-a]pyrazin-8(9H)-carboxylic acid tert-butyl ester (309 mg, 1.0 mmol) was dissolved in DCM (10 mL). HCl / Dioxane (5.0 mL, 4 M) was added under ice bath cooling, and the reaction was carried out at room temperature for 3 h. LC-MS monitoring indicated the reaction was complete. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1), yielding a pale yellow solid (300 mg, 100% yield). ESI-MS m / z: 209.1 [M+H] + .

[0223] Step 6: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6,7,8,9-tetrahydropyridino[3',4':4,5]imidazo[1,2-a]pyrazin-1-yl)thiophene-2-carboxamide

[0224] Compound 22 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0225] 1 H NMR(400MHz, DMSO-d6)δ:10.72(s,1H),9.16(s,1H),9.04(s,1H),8.40(d,J=5.4Hz,1H),7.74(s,2H),7.57(d,J=5.4 Hz,1H),6.98(s,1H),4.20(s,2H),4.15(t,J=5.3Hz,2H),3.25(d,J=5.4Hz,2H),3.10(s,3H); ESI-MSm / z:503.1[M+H] + .

[0226] Examples 23-26: Synthesis of compounds 23-26

[0227] Compounds 23-26 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 22.

[0228]

[0229]

[0230] Example 27: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6a,7,9,10-tetrahydro-6H-[1,4]oxazino[4,3-d]pyrido[3,4-b][1,4]oxazin-4-yl)thiophene-2-carboxamide (compound 27)

[0231] Step 1: Synthesis of 2-chloro-4-iodopyridin-3-ol

[0232] 2-Chloro-4-iodo-3-(methoxymethoxy)pyridine (theoretical amount, 33.4 mmol) was dissolved in DCM (100 mL), and HCl / Dioxane (4.0 M, 10 mL) was added. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by LC-MS until complete. The solution was concentrated, and EA (50 mL) was added. The pH was adjusted to approximately 6 with sodium bicarbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with EA (50 mL * 3). The organic phases were combined, concentrated, and subjected to column chromatography (DCM / MeOH = 100 / 1 to 30 / 1) to give a yellow solid (7.4 g, yield 87%). ESI-MS m / z: 255.9 [M + H]. + .

[0233] Step 2: Synthesis of (S)-3-(((4-iodo-2-chloropyridin-3-yl)oxy)methyl)morpholine-4-carboxylic acid tert-butyl ester

[0234] 2-Chloro-4-iodopyridin-3-ol (1.02 g, 4.0 mmol) and (3R)-3-(hydroxymethyl)morpholine-4-carboxylic acid tert-butyl ester (870 mg, 4.0 mmol) were dissolved in THF (30 mL), and Ph3P (1.31 g, 5.0 mmol) was added. The mixture was kept in an ice bath, and DEAD (871 mg, 5.0 mmol) was added dropwise. The mixture was stirred for 3 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. Water (50 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 5 / 1 to 1 / 1) to give a yellow solid (1.1 g, 60% yield). ESI-MS m / z: 455.0 [M+H] + .

[0235] Step 3: Synthesis of (S)-3-(((4-iodo-2-chloropyridin-3-yl)oxy)methyl)morpholine

[0236] S)-3-(((4-iodo-2-chloropyridin-3-yl)oxy)methyl)morpholine-4-carboxylic acid tert-butyl ester (1.1 g, 2.4 mmol) was dissolved in DCM (30 mL), and HCl / Dioxane (4 M, 5 mL) was added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. The solution was concentrated to give a yellow solid (1.1 g, yield over 100%), ESI-MS m / z: 355.0 [M+H]. + .

[0237] Step 4: Synthesis of (S)-4-chloro-6a,7,9,10-tetrahydro-6H-[1,4]oxazino[4,3-d]pyrido[3,4-b][1,4]oxazine

[0238] (S)-3-(((4-iodo-2-chloropyridin-3-yl)oxy)methyl)morpholine (theoretical amount, 2.4 mmol) was dissolved in Dioxane (20 mL), and Pd2(dba)3 (216 mg, 0.24 mmol), X-phos (224 mg, 0.47 mmol), and CS2CO3 (1.54 g, 4.72 mmol) were added. The mixture was stirred at 100 °C and monitored by LC-MS until the reaction was complete. Water (50 mL) was added, and the mixture was extracted with EA (30 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 5 / 1 to 1 / 1) to give a white solid (210 mg, yield 39%), ESI-MS m / z: 227.1 [M+H]. + .

[0239] Step 5: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6a,7,9,10-tetrahydro-6H-[1,4]oxazino[4,3-d]pyrido[3,4-b][1,4]oxazin-4-yl)thiophene-2-carboxamide

[0240] Compound 27 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0241] 1H NMR (400MHz, DMSO-d6) δ: 10.61 (s, 1H), 10.05 (s, 1H), 8.76 (d, J = 0.7Hz, 1H), 8.39 (d, J = 0.8Hz, 1 H),7.98(d,J=5.4Hz,1H),7.71(d,J=1.7Hz,2H),6.96(s,1H),6.84(d,J=5.6Hz,1H),4.47(dd,J =10.9,2.8Hz,1H),4.04-3.90(m,3H),3.74(d,J=12.3Hz,1H),3.59(td,J=11.6,2.4Hz,1H),3.4 7-3.38(m,1H),3.21(t,J=10.9Hz,1H),3.08(s,3H),2.97-2.86(m,1H); ESI-MSm / z:521.1[M+H] + .

[0242] Examples 28-37: Synthesis of compounds 28-37

[0243] Compounds 28-27 can be synthesized from the starting materials shown in the table using a similar method to that of compound 27.

[0244]

[0245]

[0246] Example 38: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(4H-imidazo[1,2-d]pyrido[3,4-b][1,4]oxazin-6-yl)thiophene-2-carboxamide (compound 38)

[0247]

[0248] Step 1: Synthesis of 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-carboxaldehyde

[0249] 2-Imidazole formaldehyde (2 g, 20.83 mmol) was dissolved in DMF (30 mL), and NaH (1.25 g, 31.25 mmol) and SEMCl (4.17 g, 25 mmol) were added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, and concentrated to give a yellow oil (4.8 g, yield over 100%). ESI-MS m / z: 227.1 [M+H] + .

[0250] Step 2: Synthesis of (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)methanol

[0251] 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-carboxaldehyde (4.8 g, 21.24 mmol) was dissolved in MeOH (50 mL), and NaBH4 (1.61 g, 42.48 mmol) was added. The mixture was stirred at room temperature, and the reaction was monitored by LC-MS until the starting material was completely reacted. Water (30 mL) was added, and the mixture was extracted with EA (30 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 3 / 1) to give a yellow oil (2.6 g, yield 54%). ESI-MS m / z: 229.1 [M + H]. + .

[0252] Step 3: Synthesis of 2-chloro-4-iodo-3-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)methoxy)pyridine

[0253] (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)methanol (2 g, 8.73 mmol) and 2-chloro-3-fluoro-4-iodopyridine (2.25 g, 8.73 mmol) were dissolved in DMF (50 mL), and NaH (524 mg, 13.10 mmol) was added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 3 / 1) to give a yellow solid (1.1 g, yield 27%). ESI-MS m / z: 466.0 [M + H] + .

[0254] Step 4: Synthesis of 3-((1H-imidazol-2-yl)methoxy)-2-chloro-4-iodopyridine

[0255] 2-Chloro-4-iodo-3-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)methoxy)pyridine (1.1 g, 2.37 mmol) was dissolved in DCM (20 mL), and HCl / Dioxane (4 M, 2 mL) was added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. The solution was concentrated to give a yellow solid (1.1 g, yield over 100%), ESI-MS m / z: 335.9 [M+H]. + .

[0256] Step 5: Synthesis of 6-chloro-4H-imidazo[1,2-d]pyrido[3,4-b][1,4]oxazine

[0257] 3-((1H-imidazol-2-yl)methoxy)-2-chloro-4-iodopyridine (800 mg, 2.39 mmol) was dissolved in DMSO (30 mL), and CuI (91 mg, 0.48 mmol), L-proline (110 mg, 0.96 mmol), and K₂CO₃ (660 mg, 4.78 mmol) were added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 1 / 1) to give a white solid (120 mg, yield 24%). ESI-MS m / z: 208.0 [M+H] + .

[0258] Step 6: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(4H-imidazo[1,2-d]pyrido[3,4-b][1,4]oxazin-6-yl)thiophene-2-carboxamide

[0259] Compound 38 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0260] 1 H NMR(400MHz, DMSO-d6)δ:10.68(s,1H),10.08(s,1H),8.82(d,J=1.2Hz,1H),8.61(d,J=1.2Hz,1H),8.42(d,J=5.1Hz,1H),8.06(d, J=1.2Hz,1H),7.72(dd,J=11.0,3.5Hz,3H),7.24(s,1H),6.97(t,J=1.9Hz,1H),5.56(s,2H),3.08(s,3H); ESI-MSm / z:502.0[M+H] + .

[0261] Example 39: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6H-pyrano[5,4-b:2,3-c'-]bipyridin-4-yl)thiophene-2-carboxamide (compound 39)

[0262] Step 1: Synthesis of 3-bromo-2-((tert-butyldimethylsilyl)oxy)methyl)pyridine

[0263] 2-Hydroxymethyl-3-bromopyridine (500 mg, 2.66 mmol) was dissolved in DMF (30 mL), and imidazole (468 mg, 5.32 mmol) and TBSCl (481 mg, 3.19 mmol) were added. The mixture was stirred at 60 °C and monitored by LC-MS until the reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 3 / 1) to give a yellow solid (720 mg, 90% yield). ESI-MS m / z: 302.1 [M+H] + Step 2: Synthesis of 2-(((tert-butyldimethylsilyl)oxy)methyl)-2'-chloro-3'-fluoro-3,4'-bipyridine

[0264] 3-Bromo-2-((tert-butyldimethylsilyl)oxy)methyl)pyridine (720 mg, 2.39 mmol) and 2-chloro-3-fluoropyridine-4-boronic acid (538 mg, 3.11 mmol) were dissolved in DME / H2O (30 / 3 mL). Pd(dppf)Cl2.CH2Cl2 (196 mg, 0.24 mmol) and Na2CO3 (507 mg, 4.78 mmol) were added, and the mixture was stirred at 70 °C. The reaction was monitored by LC-MS until the reactants were completely reacted. Water (30 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (DCM / MeOH = 30 / 1) to give a yellow solid (320 mg, yield 38%). ESI-MS m / z: 353.1 [M+H] + .

[0265] Step 3: Synthesis of 4-chloro-6H-pyrano[5,4-b:2,3-c'-]bipyridine

[0266] 2-(((tert-butyldimethylsilyl)oxy)methyl)-2'-chloro-3'-fluoro-3,4'-bipyridine (320 mg, 0.91 mmol) was dissolved in THF (30 mL), and TBAF (2 M, 3 mL) was added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. Water (30 mL) was added, and the mixture was extracted with EA (30 mL * 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (DCM / MeOH = 50 / 1) to give a yellow solid (170 mg, yield 86%). ESI-MS m / z: 219.0 [M + H]. + .

[0267] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(6H-pyrano[5,4-b:2,3-c'-]bipyridin-4-yl)thiophene-2-carboxamide

[0268] Compound 39 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0269] 1 H NMR (400MHz, DMSO-d6) δ: 10.67 (s, 1H), 10.05 (s, 1H), 8.83 (s, 1H), 8.62 (d, J = 5.0Hz, 2H), 8.41 (dd, J = 9.3, 6.7Hz, 2H), 7.90 (d,J=5.0Hz,1H),7.71(s,2H),7.54(dd,J=7.8,4.9Hz,1H),6.96(s,1H),5.45(s,2H),3.08(s,3H); ESI-MSm / z:513.0[M+H] + .

[0270] Example 40: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(8,9,9a,10-tetrahydro-5H,7Hpyrido[4,3-f]pyrrolo[2,1-c][1,4]oxazolidine-1-yl)thiophene-2-carboxamide (compound 40)

[0271] Step 1: Synthesis of (S)-(1-((2-bromo-3-fluoropyridin-4-yl)methyl)pyrrolidine-2-yl)methanol

[0272] 2-Bromo-3-fluoroisononaldehyde (190 mg, 0.93 mmol) and (S)-pyrrolidine-2-ylmethanol hydrochloride (256 mg, 1.86 mmol) were dissolved in MeOH (20 mL), and Et3N (282 mg, 2.79 mmol) and AcOH (112 mg, 1.86 mmol) were added. The mixture was stirred at room temperature for 2 h, and then NaBH3CN (293 mg, 4.66 mmol) was added. LC-MS monitoring showed that the reaction was complete. The solution was directly concentrated and column chromatography (DCM / MeOH = 100 / 1 to 10 / 1) to give a yellow solid (450 mg, yield over 100%), ESI-MS m / z: 289.0 [M+H]. + .

[0273] Step 2: Synthesis of (S)-1-bromo-8,9,9a,10-tetrahydro-5H,7H-pyrido[4,3-f]pyrrolo[2,1-c][1,4]oxazapine

[0274] (S)-(1-((2-bromo-3-fluoropyridin-4-yl)methyl)pyrrolidine-2-yl)methanol (450 mg, 1.48 mmol) was dissolved in THF (30 mL), and NaH (259 mg, 0.35 mmol) was added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. The solution was quenched with a small amount of methanol, concentrated, and subjected to column chromatography (DCM / MeOH = 100 / 1 to 10 / 1) to give a white solid (90 mg, yield 21%), ESI-MS m / z: 269.0 [M+H). + .

[0275] Step 3: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(8,9,9a,10-tetrahydro-5H,7Hpyrido[4,3-f]pyrrolo[2,1-c][1,4]oxazolidine-1-yl)thiophene-2-carboxamide

[0276] Compound 40 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0277] 1 H NMR(400MHz, DMSO-d6)δ:10.64(s,1H),8.78(d,J=1.0Hz,1H),8.49(d,J=1.0Hz,1H),8.31(d,J=4.6Hz,1H),7 .70(d,J=1.8Hz,2H),7.27(d,J=4.6Hz,1H),6.96(t,J=1.8Hz,1H),4.60(dd,J=12.0,2.0Hz,1H),3.91(d,J=1 3.9Hz,1H),3.68(d,J=13.7Hz,1H),3.49(dd,J=10.3,7.8Hz,1H),3.08-3.01(m,4H),2.88-2.79(m,1H),2.47 (d,J=8.5Hz,1H),1.91(dt,J=14.0,7.6Hz,1H),1.79-1.67(m,2H),1.42-1.29(m,1H), ESI-MSm / z:519.1[M+H] + .

[0278] Examples 41-55: Synthesis of compounds 41-55

[0279] Compounds 41-55 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 40.

[0280]

[0281]

[0282] Example 56: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(5-oxo-7,8,10a,11-tetrahydro-5H,10H-[1,4]oxazino[3,4-c]pyrido[3,2-f][1,4]oxazono-2-yl)thiophene-2-carboxamide (compound 56)

[0283]

[0284] Step 1: Synthesis of (R)-(6-bromo-2-fluoropyridin-3-yl)(3-(hydroxymethyl)morpholino) ketone

[0285] 6-Bromo-2-fluoro-3-pyridinecarboxylic acid (250 mg, 1.14 mmol) and (R)-3-hydroxymethylmorpholine hydrochloride (209 mg, 1.36 mmol) were dissolved in DCM (20 mL), and DIEA (441 mg, 3.42 mmol) and HATU (650 mg, 1.71 mmol) were added. The mixture was stirred at room temperature and monitored by LC-MS until the reaction was complete. The solution was directly concentrated and purified by column chromatography (DCM / MeOH = 10 / 1) to give a yellow solid (420 mg, yield over 100%), ESI-MS m / z: 319.0 [M+H]. + .

[0286] Step 2: Synthesis of (S)-2-bromo-7,8,10a,11-tetrahydro-5H,10H-[1,4]oxazino[3,4-c]pyrido[3,2-f][1,4]oxazono-5-one

[0287] The (R)-(6-bromo-2-fluoropyridin-3-yl)(3-(hydroxymethyl)morpholino) methyl ketone (420 mg, 1.14 mmol) obtained in the previous step was dissolved in THF (10 mL), and NaH (48 mg, 1.2 mmol) was added. The mixture was stirred at room temperature, and the reaction was monitored by LC-MS until the starting material was completely reacted. The mixture was quenched with a small amount of methanol, concentrated, and subjected to column chromatography (DCM / MeOH = 100 / 1 to 10 / 1) to give a white solid (160 mg, yield 50%), ESI-MS m / z: 299.0 [M+H]. + .

[0288] Step 3: Synthesis of (S)-N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(5-oxo-7,8,10a,11-tetrahydro-5H,10H-[1,4]oxazino[3,4-c]pyrido[3,2-f][1,4]oxazono-2-yl)thiophene-2-carboxamide

[0289] Compound 56 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0290] 1 H NMR (400MHz, DMSO-d6) δ: 10.66 (s, 1H), 10.05 (s, 1H), 8.72 (d, J = 1.1Hz, 1H), 8.61 (d, J = 1.2Hz, 1H), 8.56 (d,J=8.1Hz,1H),7.78(d,J=8.1Hz,1H),7.71-7.66(m,2H),6.97(t,J=1.9Hz,1H),4.55-4.43(m,2H),4.2 2(dt,J=13.5,3.5Hz,1H),4.01-3.95(m,1H),3.89(dd,J=11.5,3.4Hz,1H),3.81(dt,J=11.3,3.7Hz,1H), 3.62-3.55(m,1H),3.50(dd,J=11.4,8.2Hz,1H),3.38-3.33(m,1H),3.08(s,3H); ESI-MSm / z:549.1[M+H] + .

[0291] Examples 57-71: Synthesis of compounds 57-71

[0292] Compounds 57-71 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 56.

[0293]

[0294]

[0295]

[0296] Example 72: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(7,8-dihydro-6H-pyrrolo[2',1':2,3]imidazo[4,5-c]pyridin-3-yl)thiophene-2-carboxamide (compound 72)

[0297]

[0298] Step 1: Synthesis of 1-(6-bromo-4-nitropyridin-3-yl)pyrrolidine-2-one

[0299] Pyrrolidone-2-one (511 mg, 6.0 mmol) was dissolved in DMF (10 mL). The mixture was placed in an ice bath, and NaH (240 mg, 6.0 mmol, 60% in oil) was added. The mixture was stirred at room temperature for 15 min. Then, 2-bromo-5-fluoro-4-nitropyridine (1.1 g, 5.0 mmol) was added. The reaction mixture was heated to 50 °C and reacted for 5 h. LC-MS monitoring indicated that the reaction was complete. The solution was diluted with EA, washed with water, and the organic phase was directly concentrated and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a yellow solid (860 mg, yield 60%). ESI-MS m / z: 286.0 [M+H] + .

[0300] Step 2: Synthesis of 3-bromo-7,8-dihydro-6H-pyrrolo[2',1':2,3]imidazo[4,5-c]pyridine

[0301] 1-(6-bromo-4-nitropyridin-3-yl)pyrrolidone-2-one (860 mg, 3.0 mmol) was dissolved in AcOH (5 mL), and Fe powder (269 mg, 15 mmol) was added. The reaction was carried out at 100 °C for 5 h, and the reaction was monitored by LC-MS until the starting material was completely reacted. The mixture was concentrated, the residue EA was dissolved, and the mixture was washed with saturated sodium bicarbonate solution. The organic phase was directly concentrated and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a pale yellow solid (460 mg, yield 64%). ESI-MS m / z: 238.0 [M+H] + Step 3: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(7,8-dihydro-6H-pyrrolo[2',1':2,3]imidazo[4,5-c]pyridin-3-yl)thiophene-2-carboxamide

[0302] Compound 72 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0303] 1 H NMR(400MHz, DMSO-d6)δ:10.66(s,1H),8.76(d,J=1.1Hz,1H),8.60(d,J=1.2Hz,1H),8.56(s,1H),7.82(s,1H),7.71-7.66(m,2H),6.93(t,J =1.9Hz,1H),4.12(dt,J=13.5,3.5Hz,2H),3.99(dt,J=11.5,3.4Hz,2H),3.08(s,3H),2.67(tt,J=13.5,3.5Hz,2H); ESI-MSm / z:488.1[M+H] + .

[0304] Examples 73-75: Synthesis of compounds 73-75

[0305] Compounds 73-75 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 72.

[0306]

[0307] Example 76: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6'-tetrahydro-2H-spiro[furano[2,3-c]pyridin-3,4'-pyran]-7-yl)thiophene-2-carboxamide (compound 76)

[0308]

[0309] Step 1: Synthesis of 2-chloro-3-(3,6-dihydro-2H-pyran-4-yl)methoxy)-4-iodopyridine

[0310] 2-Chloro-4-iodopyridin-3-ol (510 mg, 2.0 mmol) and (3,6-dihydro-2H-pyran-4-yl)methanol (228 mg, 2.0 mmol) were dissolved in THF (15 mL), and Ph3P (787 mg, 3.0 mmol) was added. The mixture was placed in an ice bath, and DEAD (522 mg, 3.0 mmol) was added dropwise. The mixture was stirred overnight, and the reaction was monitored by LC-MS. The starting material showed acceptable reaction. Water (50 mL) was added, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a yellow solid (352 g, 50% yield). ESI-MS m / z: 352.0 [M+H] + .

[0311] Step 2: Synthesis of 7-chloro-2',3',5',6'-tetrahydro-2H-spiro[furano[2,3-c]pyridine-3,4'-pyran]

[0312] 2-Chloro-3-(3,6-dihydro-2H-pyran-4-yl)methoxy)-4-iodopyridine (352 mg, 1.0 mmol) was dissolved in EtOH (30 mL), and NaHCO3 (840 mg, 10 mmol) and H3PO2 (660 mg, 10 mmol) were added. The reaction was carried out at 80 °C for 2 h under argon protection. LC-MS monitoring showed that the starting material reaction was acceptable. The mixture was concentrated, and water (50 mL) was added. Extraction was performed using EA (30 mL * 2). The organic phases were combined, dried, concentrated, and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a yellow solid (120 g, yield 53%). ESI-MS m / z: 226.0 [M+H] + .

[0313] Step 3: N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6'-tetrahydro-2H-spiro[furano[2,3-c]pyridin-3,4'-pyran]-7-yl)thiophene-2-carboxamide

[0314] Compound 76 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0315] 1 H NMR(400MHz,DMSO-d6)δ:10.66(d,J=15.6Hz,1H),8.80(s,1H),8.42(s,1H) ),8.24(d,J=4.6Hz,1H),7.71(s,2H),7.41(d,J=4.6Hz,1H),6.96(s,1H),4 .74(s,2H),3.90(d,J=10.9Hz,2H),3.50(t,J=9.6Hz,2H),3.08(s,3H),1. 97(td,J=13.0,4.2Hz,2H),1.68(d,J=13.0Hz,2H),ESI-MSm / z:520.1[M+H] + .

[0316] Examples 77-91: Synthesis of compounds 77-91

[0317] Compounds 77-91 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 76.

[0318]

[0319]

[0320]

[0321] Example 92: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3'-spiro[cyclohexane-1,2'-furano[2,3-c]pyridin]-7'-yl)thiophene-2-carboxamide (compound 92)

[0322]

[0323] Step 1: Synthesis of 1-((2-chloro-3-(methoxymethoxy)pyridin-4-yl)methyl)cyclohexyl-1-ol

[0324] 2-Chloro-3-(methoxymethoxy)pyridine (1.73 g, 10 mmol) was dissolved in THF (40 mL) under argon protection and cooled to -78 °C. n-BuLi (2.5 M in hexane, 6 mL) was added dropwise. After the addition was complete, the reaction was carried out at -78 °C for 1 h. Then, a tetrahydrofuran solution (10 mL) of 1-oxaspiro[2.5]octane (2.24 g, 20 mmol, synthesis reference: Organic and Biomolecular Chemistry, 2007, vol. 5, #19, pp. 3106-3114) was added. After the addition was complete, the mixture was allowed to warm naturally to room temperature and stirred overnight. LC-MS monitoring showed that a small amount of the starting material remained in the reaction mixture. Add ice water and ethyl acetate, separate the layers, concentrate, and perform column chromatography (PE / EA = 50 / 1 to 5 / 1). Concentrate to give an off-white solid (1.43 g, 50% yield), ESI-MS m / z: 286.1 [M+H]. + .

[0325] Step 2: Synthesis of 2-chloro-4-((1-hydroxycyclohexyl)methyl)pyridin-3-ol

[0326] 1-((2-chloro-3-(methoxymethoxy)pyridin-4-yl)methyl)cyclohexyl-1-ol (1.43 g, 5.0 mmol) was dissolved in DCM (50 mL), and TFA (5 mL) was added. The reaction was allowed to proceed overnight at room temperature. LC-MS was used to monitor the reaction until complete. The mixture was directly concentrated, and the residue was dissolved in EA, washed with sodium bicarbonate aqueous solution, dried, and concentrated to give a pale yellow solid (1.2 g, 100% yield). ESI-MS m / z: 242.1 [M+H] + It can be used directly in the next reaction.

[0327] Step 3: Synthesis of 7'-chloro-3'H-spiro[cyclohexane-1,2'-furano[2,3-c]pyridine]

[0328] 2-Chloro-4-((1-hydroxycyclohexyl)methyl)pyridin-3-ol (241 mg, 1.0 mmol) was dissolved in toluene (5 mL), and phosphoric acid (70%, 0.4 mL) was added. The mixture was refluxed for 3 h. LC-MS monitoring showed the reaction was acceptable. The mixture was directly concentrated, and the residue was dissolved in EA, washed with sodium bicarbonate aqueous solution, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (100 mg, yield 45%). ESI-MS m / z: 224.1 [M+H] + .

[0329] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3'-spiro[cyclohexane-1,2'-furano[2,3-c]pyridin]-7'-yl)thiophene-2-carboxamide

[0330] Compound 92 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0331] 1 H NMR(400MHz, DMSO-d6)δ:10.63(d,J=15.6Hz,1H),8.79(s,1H),8.40(s,1H),8.22(d,J=4.6Hz,1H),7.71(s,2H),7.38( d,J=4.6Hz,1H),6.96(s,1H),3.74(s,2H),3.08(s,3H),1.97-1.89(m,4H),1.60-1.40(m,6H),ESI-MSm / z:518.1[M+H] + .

[0332] Examples 93-94: Synthesis of compounds 93-94

[0333] Compounds 93-94 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 92.

[0334]

[0335] Example 95: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-1-yl)thiophene-2-carboxamide (compound 95)

[0336]

[0337] Step 1: Synthesis of tert-butyl 2-(2-chloro-3-methylpyridin-4-yl)-2-cyanoacetate

[0338] 4.46 g (31.56 mmol) of tert-butyl cyanoacetate and 4 g (15.78 mmol) of 2-chloro-4-iodo-3-methylpyridine were dissolved in DMF (40 mL). K₂CO₃ (8.72 g, 63.12 mmol) and CuI (30 mg, 0.16 mmol) were added. The mixture was heated to 120 °C overnight under argon protection. The reaction was monitored by LC-MS until completion. The reaction mixture was diluted with water and acidified with 10% citric acid aqueous solution. Extraction was performed with EA, and the organic phases were combined, washed with brine, dried, and concentrated. Column chromatography (PE / EA = 50 / 1 to 5 / 1) yielded a pale yellow oil (1.87 g, 44%). ESI-MS m / z: 267.1 [M+H] + .

[0339] Step 2: Synthesis of 2-(2-chloro-3-methylpyridin-4-yl)acetonitrile

[0340] 1.87 g (7.0 mmol) of 2-(2-chloro-3-methylpyridin-4-yl)-2-cyanoacetate tert-butyl ester was dissolved in 5 mL of DCM, and 5 mL of TFA was added. The mixture was reacted at 40 °C for 5 h. The reaction was monitored by LC-MS until completion. The reaction solution was directly concentrated, and the residue was dissolved in DCM, washed with saturated sodium bicarbonate solution, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (833 mg, 71%). ESI-MS m / z: 167.0 [M+H] + .

[0341] Step 3: Synthesis of 4-(2-chloro-3-methylpyridin-4-yl)tetrahydro-2H-pyran-4-onitrile

[0342] 2-(2-chloro-3-methylpyridin-4-yl)acetonitrile (833 mg, 5.0 mmol) was dissolved in THF (30 mL). The mixture was kept in an ice bath. HMDSNa (5.5 mL, 1.0 M in THF) was added, and the mixture was allowed to react in an ice bath for 30 min. The ice bath was then removed, and the mixture was allowed to react at room temperature for 30 min. 1-Iodo-2-(2-iodoethoxy)ethane (1.8 g, 5.5 mmol) was added, and the mixture was allowed to react at room temperature for 30 min. HMDSNa (5.5 mL, 1.0 M in THF) was added again, and the mixture was allowed to react overnight at room temperature. The reaction was monitored by LC-MS until complete. The reaction was quenched with ammonium chloride aqueous solution. The mixture was diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (473 mg, 40% yield). ESI-MS m / z: 237.1 [M+H] + .

[0343] Step 4: Synthesis of 1-chloro-2',3',5',6'-tetrahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-6(7H)-one

[0344] 4-(2-chloro-3-methylpyridin-4-yl)tetrahydro-2H-pyran-4-onitrile (473 mg, 2.0 mmol) was dissolved in THF (20 mL), cooled to -78 °C, and LDA (2.0 mL, 2.0 M) was added. After the addition was complete, the system was kept at -78 °C for 3 h. The reaction was monitored by LC-MS until completion. The reaction was quenched by adding hydrochloric acid aqueous solution (2 M), stirred at room temperature for 30 min, water was added, EA was added, the mixture was separated, the organic phase was concentrated, and column chromatography was performed (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (356 mg, yield 75%). ESI-MS m / z: 238.1 [M+H] + .

[0345] Step 5: Synthesis of 1-chloro-2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridine-5,4'-pyran]-6-ol

[0346] 1-Chloro-2',3',5',6'-tetrahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-6(7H)-one (356 mg, 1.5 mmol) was dissolved in MeOH (2.5 mL) and THF (5.0 mL). NaBH4 (114 mg, 3.0 mmol) was added under ice bath conditions, and the mixture was stirred at room temperature for 2 hours. LC-MS monitoring showed acceptable reaction conditions. The reaction solution was quenched with water (50 mL), extracted with EA (30 mL x 2), and the organic phases were combined, dried over Na2SO4, and concentrated to obtain a colorless gel (360 mg, 100%). The crude product was used directly in the next reaction step.

[0347] Step 6: Synthesis of O-(1-chloro-2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-6-yl)O-phenylthiocarbamate

[0348] 1-Chloro-2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridine-5,4'-pyran]-6-ol (theoretical amount, 1.5 mmol) was dissolved in ACN, DMAP was added, and the reaction was carried out at room temperature for 5 h. The reaction was monitored by LC-MS until completion. The reaction solution was quenched with water (50 mL), extracted with EA (30 mL * 2), the organic phases were combined, dried over Na2SO4, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (376 mg, yield 67%). ESI-MS m / z: 376.1 [M + H] + .

[0349] Step 7: Synthesis of 1-chloro-2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridine-5,4'-pyran]

[0350] O-(1-chloro-2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-6-yl)O-phenyl thiocarbamate (376 mg, 1.0 mmol) was dissolved in toluene (10 mL), AIBN (82 mg, 0.5 mmol) was added, followed by Et3SiH (3 mL). The mixture was heated to 80 °C under argon protection for 3 h. The reaction was monitored by LC-MS until completion. The solution was directly concentrated and subjected to column chromatography (PE / EA = 50 / 1 to 5 / 1) to give a pale yellow solid (120 mg, yield 54%). ESI-MS m / z: 224.1 [M+H] + .

[0351] Step 8: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6,6',7-hexahydrospiro[cyclopenta[c]pyridin-5,4'-pyran]-1-yl)thiophene-2-carboxamide

[0352] Compound 95 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0353] 1 H NMR(400MHz, DMSO-d6)δ:10.62(d,J=15.6Hz,1H),8.76(s,1H),8.39(s,1H),8.24(d,J=4.6Hz,1H),7.71-7.66(m,2H),7.38(d,J=4.6H z,1H),6.96(s,1H),3.74-3.66(m,2H),3.62-3.56(m,2H),3.08(s,3H),2.98-2.80(m,2H),1.97-1.83(m,6H); ESI-MSm / z:518.1[M+H] + .

[0354] Examples 96-103: Synthesis of compounds 96-103.

[0355] Compounds 96-103 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 95.

[0356]

[0357]

[0358] Example 104: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(1'-methyl-1',2,2',3,5,6-hexahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridin]-7'-yl)thiophene-2-carboxamide (compound 104)

[0359] Step 1: Synthesis of 7'-chloro-2,3,5,6-tetrahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridine]-2'(1'H)-one

[0360] 7-Chloro-1,3-dihydro-2H-pyrrolo[2,3-c]pyridin-2-one (843 mg, 5.0 mmol) was dissolved in THF (30 mL), cooled to -78 °C, and HMDSLi (15 mL, 1.0 M in THF) was added dropwise over approximately 20 min. After the addition was complete, the temperature was slowly raised to 0 °C, and 1-iodo-2-(2-iodoethoxy)ethane (1.8 g, 5.5 mmol) was added. The reaction was allowed to proceed at room temperature for 30 min, and then refluxed overnight. LC-MS monitoring showed a satisfactory reaction. The reaction was quenched with ammonium chloride aqueous solution, diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1) to give a pale yellow solid (525 mg, yield 44%). ESI-MS m / z: 239.1 [M+H] + .

[0361] Step 2: Synthesis of 7'-chloro-1'-methyl-2,3,5,6-tetrahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridine]-2'(1'H)-one

[0362] 7'-Chloro-2,3,5,6-tetrahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridine]-2'(1'H)-one (525 mg, 2.2 mmol) was dissolved in DMF (10 mL), placed in an ice bath, and NaH (106 mg, 2.64 mmol) was added. The mixture was stirred for 30 min, and MeI (344 mg, 2.42 mmol) was added. The reaction was allowed to proceed at room temperature for 5 h. LC-MS monitoring showed that the reaction was acceptable. The reaction was quenched by adding ammonium chloride aqueous solution, diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1) to give a pale yellow solid (390 mg, 70% yield). ESI-MS m / z: 253.1 [M+H] + Step 3: Synthesis of 7'-chloro-1'-methyl-1',2,2',3,5,6-hexahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridine]

[0363] 7'-Chloro-1'-methyl-2,3,5,6-tetrahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridine]-2'(1'H)-one (390 mg, 1.54 mmol) was dissolved in BH3·THF solution (2.0 M, 10 mL), and the mixture was refluxed overnight. LC-MS monitoring showed the reaction was acceptable. The reaction was quenched with MeOH, and the mixture was refluxed again for 1 h. The mixture was concentrated, and the residue EA was dissolved, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1) to give a pale yellow solid (240 mg, yield 65%). ESI-MS m / z: 239.1 [M+H] + .

[0364] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(1'-methyl-1',2,2',3,5,6-hexahydrospiro[pyran-4,3'-pyrrolo[2,3-c]pyridin]-7'-yl)thiophene-2-carboxamide

[0365] Compound 104 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0366] 1 H NMR(400MHz, DMSO-d6)δ:10.64(d,J=15.6Hz,1H),8.79(s,1H),8.38(s,1H),8.22(d,J=4.6Hz,1H),7.71-7.66(m,2H),7.36(d,J=4.6Hz,1H) ,6.96(s,1H),3.74-3.66(m,2H),3.62-3.56(m,2H),3.12-3.03(m,2H) ,3.08(s,3H),2.71(s,3H),1.97-1.83(m,4H);ESI-MSm / z:533.1[M+H] + .

[0367] Examples 105-107: Synthesis of compounds 105-107

[0368] Compounds 105-107 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 104.

[0369]

[0370] Example 108: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridin-1,4'-pyran]-4-yl)thiophene-2-carboxamide (compound 108)

[0371]

[0372] Step 1: Synthesis of 4-chloro-2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridin-1,4'-pyran]-3-one

[0373] 2-Chloro-4-iodonicotinic acid methyl ester (1.49 g, 5.0 mmol) was dissolved in THF (50 mL), cooled to -78 °C, and i-PrMgCl·LiCl (5 mL, 1.3 M in THF) was added dropwise over approximately 20 min. After the addition was complete, the system was maintained at -78 °C for 1 h. Tetrahydro-4H-pyran-4-one (550 mg, 5.5 mmol) was added, and the reaction was allowed to proceed for 30 min. The cooling device was then removed, and the system was slowly raised to room temperature for 1 h. LC-MS monitoring showed that the reaction was acceptable. The reaction was quenched by adding ammonium chloride aqueous solution, diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1) to give a pale yellow solid (480 mg, yield 40%). ESI-MS m / z: 240.0 [M+H] + .

[0374] Step 2: Synthesis of 4-chloro-2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridine-1,4'-pyran]-3-ol

[0375] 4-Chloro-2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridin-1,4'-pyran]-3-one (480 mmol, 2.0 mmol) was dissolved in DCM (15 mL) under argon protection in an ice bath. DIBAL-H (6 mL, 1 M in hexane) was added dropwise. After addition, the reaction was allowed to proceed at room temperature for 2 h. The reaction was monitored by LC-MS until complete. The reaction was quenched by adding ammonium chloride aqueous solution. The DCM was diluted, separated, washed with water, dried, and concentrated to give a pale yellow solid (482 mg, 100% yield). ESI-MS m / z: 242.0 [M+H] + It can be used directly in the next reaction.

[0376] Step 3: Synthesis of 4-chloro-2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridine-1,4'-pyran].

[0377] The 4-chloro-2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridine-1,4'-pyran]-3-ol obtained in the previous step (theoretical amount, 2.0 mmol) was dissolved in TFA (10 mL), placed in an ice bath, and Et3SiH (2 mL) was added. The reaction was allowed to proceed at room temperature for 1 h. The reaction was monitored by LC-MS until completion. The mixture was directly concentrated, and the residue was added to DCM, neutralized with saturated sodium bicarbonate aqueous solution, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1) to give a pale yellow solid (226 mg, yield 50%). ESI-MS m / z: 226.1 [M+H] + .

[0378] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2',3',5',6'-tetrahydro-3H-spiro[furano[3,4-c]pyridin-1,4'-pyran]-4-yl)thiophene-2-carboxamide

[0379] Compound 108 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0380] 1 H NMR(400MHz, DMSO-d6)δ:10.66(s,1H),8.71(d,J=1.4Hz,1H),8.62(d,J=4.9Hz,1H), 8.12(d,J=1.4Hz,1H),7.69(d,J=2.0Hz,2H),7.44(d,J=4.9Hz,1H),6.97(t,J=2.0Hz ,1H),5.37(s,2H),3.87(dd,J=10.6,5.3Hz,2H),3.72(td,J=11.8,2.1Hz,2H),3.08( s,3H),2.03(td,J=13.1,5.2Hz,2H),1.64(d,J=13.2Hz,2H); ESI-MSm / z:520.1[M+H] + .

[0381] Examples 109-117: Synthesis of compounds 109-117

[0382] Compounds 109-117 can be synthesized from the starting materials shown in the table using a similar method to that of compound 108.

[0383]

[0384]

[0385] Example 118: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2'-methyl-2',3'-dihydrospiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-4'-yl)thiophene-2-carboxamide (compound 118)

[0386]

[0387] Step 1: Synthesis of 4'-bromo-2'-(4-methoxybenzyl)spiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one

[0388] 4-Bromo-2-(4-methoxybenzyl)-1,2-dihydro-3H-pyrrolo[3,4-c]pyridin-3-one (1.33 g, 4.0 mmol, synthesis reference WO2019204550) was dissolved in DMF (15 mL), and the mixture was kept under argon protection and cooled to 0 °C on an ice bath. NaH (480 mg, 12.0 mmol, 60% in oil) was added, and the mixture was stirred at 0 °C for 30 min. 1,5-Diiodopentane was then added, and the mixture was allowed to react overnight at room temperature. LC-MS monitoring showed a satisfactory reaction. The reaction was quenched with water, diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1) to give a pale yellow solid (800 mg, 50% yield). ESI-MS m / z: 401.1 [M+H] + .

[0389] Step 2: Synthesis of 4'-bromospiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one

[0390] 4'-Bromo-2'-(4-methoxybenzyl)spiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one (800 mg, 2.0 mmol) was dissolved in ACN / H2O (2 / 1, 15 mL), placed in an ice bath, and cerium ammonium nitrate (3.29 g, 6.0 mmol) was added. The reaction was carried out in an ice bath for 1 h, and then overnight at room temperature. LC-MS monitoring showed a satisfactory reaction. The mixture was diluted with ethyl acetate, separated, washed with water (EA phase), dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1) to give a pale yellow solid (365 mg, 65% yield). ESI-MS m / z: 281.0 [M+H] + .

[0391] Step 3: Synthesis of 4'-bromo-2'-methylspiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one

[0392] 4'-Bromospiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one (365 mg, 1.3 mmol) was dissolved in DMF (10 mL), placed in an ice bath, and NaH (62 mg, 1.56 mmol) was added. The mixture was stirred for 30 min, and MeI (222 mg, 1.56 mmol) was added. The reaction was allowed to proceed at room temperature for 5 h. LC-MS monitoring showed that the reaction was acceptable. The reaction was quenched by adding ammonium chloride aqueous solution, diluted with EA, separated, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 2 / 1) to give a pale yellow solid (295 mg, yield 77%). ESI-MS m / z: 295.0 [M+H] + .

[0393] Step 4: Synthesis of 4'-bromo-2'-methyl-2',3'-dihydrospiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]

[0394] 4'-Bromo-2'-methylspiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridine]-3'(2'H)-one (295 mg, 1.0 mmol) was dissolved in BH3·THF solution (2.0 M, 10 mL), and the mixture was refluxed overnight. LC-MS monitoring showed the reaction was acceptable. The reaction was quenched with MeOH, and the mixture was refluxed again for 1 h. The mixture was concentrated, and the residue EA was dissolved, washed with water, dried, concentrated, and subjected to column chromatography (PE / EA = 50 / 1 to 1 / 1) to give a pale yellow solid (140 mg, 50% yield). ESI-MS m / z: 281.1 [M+H] + .

[0395] Step 5: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2'-methyl-2',3'-dihydrospiro[cyclohexane-1,1'-pyrrolo[3,4-c]pyridin]-4'-yl)thiophene-2-carboxamide

[0396] Compound 118 was obtained by a similar Suzuki coupling reaction as in Example 1.

[0397] 1 H NMR(400MHz, DMSO-d6)δ:10.66(d,J=15.6Hz,1H),8.80(s,1H),8.41(s,1H),8.26(d,J=4.6Hz,1H),7.71-7.65(m,2H),7.41(d,J =4.6Hz,1H),6.97(s,1H),3.70(s,2H),3.08(s,3H),2.33(s,3H),1.97-1.89(m,4H),1.60-1.40(m,6H),ESI-MSm / z:531.1[M+H] + .

[0398] Examples 119-121: Synthesis of compounds 119-121

[0399] Compounds 119-121 can be synthesized from the starting materials shown in the table using a similar method to that of compound 118.

[0400]

[0401]

[0402] Example 122: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2H-spiro[furano[2,3-c]pyridin-3,2'-[1,3]dithiacyclopentane]-7-yl)thiophene-2-carboxamide (compound 122)

[0403]

[0404] Step 1: Synthesis of 6-oxide of 3-oxo-2,3-dihydrofurano[2,3-c]pyridine

[0405] Furano[2,3-c]pyridin-3(2H)-one (676 mg, 5.0 mmol, synthesis reference WO2013014587) was dissolved in DCM (20 mL), and m-CPBA (1.32 g, 7.5 mmol, 85%) was added. The reaction was carried out overnight at room temperature. LC-MS monitoring showed that the reaction was complete. The reaction solution was filtered, the filtrate was collected, concentrated, and column chromatography was performed (PE / EA = 10 / 1 to 1 / 1). The concentrate yielded a white solid (453 mg, yield 60%), ESI-MS m / z: 152.0 [M+H]. + .

[0406] Step 2: Synthesis of 7-bromofurano[2,3-c]pyridine-3(2H)-one

[0407] 3-O-2,3-dihydrofurano[2,3-c]pyridine 6-oxide (453 mg, 3.0 mmol) was dissolved in dry DCM (300 mL, adjusted to approximately 0.01 M), TBAB (1.45 g, 4.5 mmol) was added, followed by powdered molecular sieve (5.0 g). The mixture was stirred at room temperature for approximately 10 mins, then p-toluenesulfonic anhydride (1.47 g, 4.5 mmol) was added, and the mixture was stirred overnight at room temperature. LC-MS monitoring indicated that the reaction was complete. The solution was filtered, concentrated, and subjected to column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a pale yellow solid (320 mg, 50% yield). ESI-MS m / z: 214.0 [M+H] + .

[0408] Step 3: Synthesis of 7-bromo-2H-spiro[furano[2,3-c]pyridine-3,2'-[1,3]dithiopentane].

[0409] 7-Bromofurano[2,3-c]pyridin-3(2H)-one (320 mg, 1.5 mmol) was dissolved in chloroform (5 mL), and ethane-1,2-dithiol (424 mg, 4.5 mmol) and boron trifluoride diethyl ether (426 mg, 3.0 mmol) were added. The mixture was refluxed under argon protection for 4 h, and the reaction was monitored by LC-MS, showing acceptable reaction. After cooling, the reaction solution was washed with NaOH aqueous solution (3 M, 20 mL), washed with water, concentrated, and subjected to column chromatography (PE / EA = 20 / 1 to 2 / 1) to give a pale yellow solid (290 mg, yield 66%), ESI-MS m / z: 289.9 [M+H]. + .

[0410] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(2H-spiro[furano[2,3-c]pyridin-3,2'-[1,3]dithiacyclopentane]-7-yl)thiophene-2-carboxamide

[0411] Compound 122 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0412] 1 H NMR (400MHz, DMSO-d6) δ: 10.65 (s, 1H), 8.70 (d, J = 1.4Hz, 1H), 8.62 (d, J = 4.9Hz, 1H), 8.12 (d, J = 1.4Hz, 1H), 7.70 (d, J = 2.0Hz, 2H), 7.44 (d, J = 4 .9Hz,1H),6.97(t,J=2.0Hz,1H),5.17(s,2H),2.87(dd,J=10.6,5.3Hz,2H),2.72(td,J=11.8,2.1Hz,2H),3.08(s,3H); ESI-MSm / z:540.0[M+H] + .

[0413] Examples 123-129: Synthesis of compounds 123-129

[0414] Compounds 123-129 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 122.

[0415]

[0416]

[0417] Example 130: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3-((tetrahydro-4H-pyran-4-ylidene)methyl)pyridin-2-yl)thiophene-2-carboxamide (compound 130)

[0418]

[0419] Step 1: Synthesis of 3-(bromomethyl)-2-chloropyridine

[0420] (2-chloropyridin-3-yl)methanol (4.3 g, 30 mmol) was dissolved in DCM (150 mL). The mixture was placed in an ice bath, and a solution of PBr3 (16.2 g, 60 mmol) in DCM (20 mL) was added dropwise. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by LC-MS until complete. The solution was diluted with DCM, washed with saturated sodium bicarbonate solution, washed with water, dried, and concentrated to give a pale yellow oil (6.5 g, 100% yield). ESI-MS m / z: 205.9 [M+H] + .

[0421] Step 2: Synthesis of diethyl (2-chloropyridin-3-yl)methyl)phosphonate

[0422] 3-(bromomethyl)-2-chloropyridine (2.06 g, 10 mmol) and toluene (10 mL) were reacted overnight at 100 °C under argon protection. The reaction was monitored by LC-MS until completion. The solution was directly concentrated and subjected to column chromatography (PE / EA = 20 / 1 to 1 / 1) to give a pale yellow gel (2.0 g, yield 76%), ESI-MS m / z: 264.1 [M+H]. + .

[0423] Step 3: Synthesis of 2-chloro-3-((tetrahydro-4H-pyran-4-ylidene)methyl)pyridine

[0424] Diethyl (2-chloropyridin-3-yl)methylphosphonate (1.27 g, 4.8 mmol) was dissolved in THF (30 mL). Under argon protection, the solution was cooled to -78 °C, and t-BuOK (539 mg, 4.8 mmol) was added. The mixture was stirred for 30 min, and then a THF solution of tetrahydro-4H-pyran-4-one (400 mg, 4.0 mmol) (5 mL) was added. After the addition was complete, the mixture was slowly heated to room temperature and reacted for 5 h. The reaction was monitored by LC-MS until it was complete. The solution was directly concentrated and column chromatography (PE / EA = 20 / 1 to 5 / 1) to give a pale yellow gel (600 g, yield 71%). ESI-MS m / z: 210.1 [M+H] + .

[0425] Step 4: Synthesis of N-(3-chloro-5-(methylsulfonamido)phenyl)-4-(3-((tetrahydro-4H-pyran-4-ylidene)methyl)pyridin-2-yl)thiophene-2-carboxamide

[0426] Compound 130 was obtained according to a similar Suzuki coupling reaction as in Example 1.

[0427] 1 H NMR (400MHz, DMSO-d6) δ: 10.60 (s, 1H), 10.08 (s, 1H), 8.60-8.54 (m, 2H), 8.16 (d, J = 1. 4Hz, 1H), 7.69 (dt, J=6.5, 1.9Hz, 2H), 7.65 (dd, J=7.8, 1.7Hz, 1H), 7.37 (dd, J=7.7, 4. 7Hz,1H),6.96(t,J=1.9Hz,1H),6.41(s,1H),3.71(t,J=5.4Hz,2H),3.51(t,J=5.5Hz, 2H),3.08(s,4H),2.39(t,J=5.4Hz,2H),2.22(t,J=5.4Hz,2H); ESI-MSm / z:504.1[M+H] + .

[0428] Examples 131-140: Synthesis of compounds 131-140

[0429] Compounds 131-140 can be synthesized from the starting materials shown in the table using a similar method to that used for compound 130.

[0430]

[0431] Example 141: Screening for antiproliferative activity of HCT-116 cells

[0432] This study analyzed the cytotoxicity of HCT-116 colon cancer cells after six days of treatment with a DHX9 enzyme inhibitor using a cell proliferation assay. The HCT-116 cell line was purchased from Wuhan Pronosei Biotechnology Co., Ltd., and cultured in McCoy's 5A medium (Viva cell) at 37°C in a Thermo cell culture incubator with 5% CO2. In this experiment, the DHX9 enzyme inhibitor was dissolved in DMSO to an initial concentration of 10 μM, and then triple-diluted in DMSO, for a total of eight data points. The final concentration of DMSO was 0.5%.

[0433] HCT-116 cells were seeded into white 96-well plates, with 80 μL of cell suspension per well, containing 250 HCT-116 cells. The plates were incubated overnight in a CO2 incubator. Then, 20 μL of different concentrations of DHX9 enzyme inhibitor solution were added to each well, and the plates were incubated for six days. A separate cell plate was prepared, and the signal value on the day of drug addition was recorded as the maximum value (Max value in the equation below) for data analysis. 25 μL of chemiluminescent cell viability assay reagent (Novazia, DD1101) was added to each well of this cell plate, and the plate was incubated at room temperature for 10 minutes to stabilize the luminescence signal. Readings were performed using a multi-label analyzer.

[0434] The original data were converted into inhibition rate using the equation (Sample-Min) / (Max-Min)*100%, IC 50 The values ​​can be obtained by curve fitting using four parameters (obtained in the "log(inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Min: cell wells treated with 0.5% DMSO; Max: cell-free blank control wells. Table 2 provides the inhibitory activity of some compounds of the present invention on the proliferation of HCT-116 cells. Screening results are shown in Table 1.

[0435] Table 1: Antiproliferative activity of compounds against HCT-116 cells (IC50) 50 )

[0436]

[0437] As can be seen from the data in the table above, the compounds of this invention have strong anti-proliferative activity against HCT-116 cells, especially compounds 76, 79, and 108, which have high IC50 values. 50 The value is less than 50 nM.

[0438] Example 142: Screening of DXH9 enzyme activity

[0439] This study evaluated the in vitro inhibition of DHX9 protease activity by the compounds of this invention. The DHX9 protease inhibitors were characterized using the ADP-Glo ​​(Promega, V9103) kinase assay at 5 μM ATP (Promega). In this experiment, the final concentration of DHX9 (150-1150, ICE, S2309T-H13HF) protease was 0.25 nM, the final concentration of substrate SB-RNA (ICE, DRU-102) was 2.5 nM, and the final concentration of DMSO (Sigma, D4540) was 1%.

[0440] The compound was dissolved in DMSO to obtain a 10 mM stock solution. A dose-gradient reaction solution was prepared with an initial compound concentration of 3 μM, followed by a three-fold dilution in DMSO for a total of ten data points. Using an ECHO ultrasonic pipetting apparatus (LABCYTE, 655), 0.1 μL of the diluted compound solution was transferred to a 384-well microplate (Revvity, 6007299). 5 μL of LDHX9 (150-1150) enzyme solution was added to each well of the 384-well microplate, and the plates were incubated at 25°C for 10 minutes. Wells containing 1% DMSO and the enzyme served as high-value controls, and wells containing 1% DMSO and buffer served as low-value controls. 5 μL of SB-RNA and ATP solution were added to each well, and the plates were incubated at 25°C for 60 minutes. 5 μL of ADP-GloRegent solution was added to each well, and the plates were incubated at 25°C for 40 minutes. Add 10 μL of ADP-Glo ​​Detection solution to each well and incubate at 25°C for 40 minutes. Read the RLU (Relative Luminescence Unit) signal using a BMG (PHERAstar FSX) microplate reader.

[0441] The inhibition rate of the compound on DHX9 protein activity was calculated using the following formula: Inhibition percentage (%) = 100 * (mean value of DMSO group - mean value of compound) / (mean value of DMSO group - mean value of blank control group). The IC50 of the compound was then fitted using a nonlinear regression equation with XLfit 5.5.0 software. 50 Curves. Table 1 shows the inhibitory activity of some compounds of the present invention against DHX9 protein. Screening results are shown in Table 2.

[0442] Table 2: Inhibitory activity of compounds against DHX9 enzyme (IC50) 50 )

[0443]

[0444] As can be seen from the data in the table above, the compounds of this invention have strong DHX9 enzyme inhibitory activity, especially compounds 76, 79, and 108, which have IC50 values ​​of [missing information]. 50 The value is less than 2nM.

[0445] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound, or an isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate thereof, having a structure as shown in formula (1): In formula (1): m is 0, 1, or 2; Ring A is a 5-membered heteroaryl group; R 1 Selected from C 1-6 Alkyl or C 3-6 cycloalkyl; R 2 Selected from halogen or C 1-3 alkyl; R 3 Selected from halogens, CN, C 1-3 Alkyl, C 3-6 cycloalkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, hydroxylated C 1-3 Alkyl, cyano-substituted C 1-3 Alkyl or C 1-3 Alkoxy-substituted C 1-3 alkyl; Ring B is selected from: in, Indicates a single bond or a double bond; n is 0, 1, 2, 3 or 4; The ring C is a 4-8 membered heterocyclic alkyl or a 5-8 membered heteroaryl; Ring D is C 3-8 Cycloalkylene or 4-8 membered heterocycloalkylene; Ring E is C 3-12 Cycloalkylene or 4-12 membered heterocyclic alkylene; G 1 and G 2 Each is independently selected from the following groups: chemical bond, O, C(O), CH2, -CH2CH2-, -CH2O- or -OCH2-, S, NR i or * indicates the location linked to ring C, G 1 and G 2 They are not both chemical bonds; G 3 and G 4 Each is independently selected from the following groups: chemical bond, O, C(O), CH2, S, or NR. i G 3 and G 4 They are not both chemical bonds; G 5 Select from the following group: chemical bond, O, C(O) or CH2; Each R 5 Independently selected from the following groups: H, halogens, CN, C 1-6 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl, cyano-substituted C 1-3 Alkyl or C 1-3 Alkoxy-substituted C 1-3 Alkyl; or, two Rs 5 It forms a -C(O)- with the same carbon atom it is attached to; R a R b and R c Independently selected from: H, halogen, C 1-6 Alkyl, C 1-3 Alkoxy, C 1-3 Halogenated alkyl or C 1-3 Alkoxy-substituted C 1-3 alkyl; R d and R e Each is independently selected from the following group: substituted or unsubstituted C 1-6 Alkyl, or substituted or unsubstituted C 3-6 cycloalkyl; the substitution refers to having one or more substituents selected from the group consisting of: halogen, CN, OH, C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 1-3 Halogenated alkoxy groups; or R d and R e Together with the attached S atom, it forms a 3-7 member monocyclic heterocyclic alkyl group or a 4-10 member bicyclic heterocyclic alkyl group; R f Selected from the following groups: H, halogens, CN, C 1-6 Alkyl or C 1-3 Halogenated alkyl groups; R i Selected from the following groups: H, C 1-6 Alkyl or C 1-3 Halogenated alkyl groups.

2. The compound of claim 1, R 1 Selected from the following group: Me, Et, 3. The compound of claim 1, R 2 Select from the following group: Me, Et, F, Cl, or Br.

4. The compound of claim 1, R 3 Selected from the following groups: halogens, CN, Me, Et, CF3, CHF2, OMe, OEt, CH2OH, CH2OMe, CH2CN.

5. The compound of claim 1, wherein ring A is selected from the group consisting of thienyl, pyrroleyl, pyrazolyl, furanyl, isothiazolyl, or imidazolyl.

6. The compound of claim 1, wherein ring B is selected from the group consisting of: Where m, n, ring C, ring D, ring E, and R are... 5 R a R b R c R d R e R f and R i The definition is as described in claim 1.

7. The compound of claim 1, wherein ring B is selected from the group consisting of:

8. The compound of claim 1, wherein the compound has a structure selected from the group consisting of:

9. A pharmaceutical composition for treating, modulating, and / or preventing DHX9-mediated related diseases, characterized in that, The pharmaceutical composition comprises: The active ingredient is any one of the compounds, isomers, polymorphs, pharmaceutically acceptable salts, hydrates or solvates as described in any one of claims 1-8; and Optional pharmaceutically acceptable excipients or carriers.

10. Use of a compound, or an isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate thereof, as described in any one of claims 1-8, or a pharmaceutical composition as described in claim 9, characterized in that, Used to prepare drugs for the prevention and / or treatment of diseases mediated by DHX9.

11. A method for treating, modulating, and / or preventing DHX9-mediated related diseases, characterized in that, The method includes the step of administering to a desired individual the compound, or an isomer, polymorph, pharmaceutically acceptable salt, hydrate, or solvate thereof, as described in any one of claims 1-8, or the pharmaceutical composition as described in claim 9.

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