Substituted fused heterocyclic compound and its preparation method and application

Abstract

A substituted fused heterocyclic compound and its preparation method and application. A compound having structural formula (I) or a pharmaceutically acceptable salt thereof is provided. The compound or the pharmaceutically acceptable salt thereof can be used as an ATR inhibitor and has high activity.

Description

Technical Field

[0001] The present invention relates to a substituted fused heterocyclic compound and a preparation method and application thereof. Background Art

[0002] In each cell cycle, human cells must accurately and efficiently replicate the entire genome. This is a very challenging and complex project because it means replicating more than 6 billion DNA fragments (bases), and the accuracy and efficiency requirements are extremely high. In this process, there will inevitably be many obstacles that prevent or affect the normal replication of DNA. Therefore, normal human cells have a powerful response mechanism to ensure that the entire genome is accurately replicated once in each cell cycle (Saldivar JCet al, 2017, Nat. Rev. Mol. Cell Biol. 18, 622-636).

[0003] There are many regulatory molecules in human cells that are involved in maintaining the stability and integrity of the genome, one of which is the ataxia telangiectasia mutated gene and Rad3-related (ATR) protein kinase. When intracellular DNA damage or replication stress occurs, ATR is recruited to the appropriate location and then activated. The activated ATR plays an important regulatory role in many aspects, including arresting the cell cycle, inhibiting replication origins, promoting the synthesis of deoxynucleotides, initiating replication forks, and so on.

[0004] Unlike normal cells, cancer cells have an unstable genome due to defects in DNA damage response and / or increased replication pressure, which is a key hallmark of cancer. These characteristics of cancer cells, which are different from normal cells, make it easier for cancer cells to accumulate gene mutations, rearrangements, copy number changes and other events during division, which may promote the clonal evolution of cancer cells; but at the same time, these characteristics are also defects that cause cancer cell-specific vulnerabilities. Taking full advantage of these vulnerabilities is expected to enhance the effectiveness of anti-cancer treatment and thus improve the prognosis of cancer patients (Pilié. PGetal, 2019, Nat. Rev. Clin. Oncol. 16, 81-104).

[0005] Cancer cells are more dependent on ATR than normal cells, which is one of the above vulnerabilities. Given that oncogene-induced replication stress is the main source of cancer genome instability, the survival of cancer cells becomes highly dependent on a sophisticated replication stress response and is therefore susceptible to ATR inhibitors. Early work in cell lines and mouse models has provided preclinical support for this concept. In addition, there is evidence that mice with reduced ATR levels are highly resistant to the development of cancer. These results highlight the potential of targeting ATR in cancer treatment. Therefore, the development of highly active and well-drugable ATR inhibitors is expected to provide humans with a new weapon to fight cancer (Lecona, E. et al, 2018, Nat. Rev. Cancer 18, 586–595).

[0006] Prior art WO2014089379A1, WO2011154737A1, WO2016020320A1, etc. disclose compounds with ATR kinase inhibitory activity. Several ATR kinase inhibitors have entered the clinical trial stage, including:

[0007]

[0008] In addition, the prior art (ACS Med. Chem. Lett. 2015, 6, 42-46) also discloses the following compounds having ATR kinase inhibitory activity:

[0009]

[0010] However, to date, no ATR kinase inhibitor has been marketed, and thus new ATR inhibitors are still urgently needed in the art, especially ATR inhibitors with high activity and good drugability. Summary of the invention

[0011] One object of the present invention is to provide a novel compound as an ATR inhibitor.

[0012] Another object of the present invention is to provide a method for preparing the compound.

[0013] Another object of the present invention is to provide the use of the compound.

[0014] Another object of the present invention is to provide a pharmaceutical composition comprising the compound and use thereof.

[0015] Another object of the present invention is to provide an intermediate for preparing the compound.

[0016] Another object of the present invention is to provide a method for preparing the intermediate.

[0017] In one aspect, the present invention provides a compound having structural formula I or a pharmaceutically acceptable salt thereof,

[0018]

[0019] in:

[0020] Y 1 , Y 2 , Y 3 , Y 4 are each independently CH or N, and Y 1 and Y 2 At least one of them is N, Y 3 and Y 4 At least one of them is CH;

[0021] R 1 is a 5-membered and 6-membered heteroaromatic ring group or a 6-membered and 6-membered heteroaromatic ring group containing n heteroatoms, or a 5-membered or 6-membered aromatic ring group containing m heteroatoms, wherein n is selected from an integer of 1-4, m is selected from an integer of 0-3, and the heteroatoms are selected from one or two of O, N, and S; in the 5-membered and 6-membered heteroaromatic ring group or the 6-membered and 6-membered heteroaromatic ring group containing n heteroatoms, the heteroaromatic ring group is optionally selected from halogen, -OH, -OMe, -NH2, -NHMe, -NMe2, -C N, -CONH2, -Me, or one or more substituents thereof; in the 5-membered or 6-membered aromatic ring group containing m heteroatoms, the aromatic ring group is optionally substituted by one or more substituents selected from 4-6-membered heterocycloalkyl, halogen, -OH, -OMe, -NH2, -NHMe, -NMe2, -CN, -CONH2, or -Me; when the aromatic ring groups are substituted by 4-6-membered heterocycloalkyl, they may be connected by a single bond or may form a cyclic ring with each other;

[0022] R 2 -OH, C 1-6 Alkoxy, -NR 3 R 4 、-CN、C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl, the heteroatom is selected from one of O and N; the C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl is optionally selected from halogen, -OH, C 1-6 Alkoxy, -NR 5 R 6 , substituted by one or more substituents in -CN;

[0023] R3 , R 4 are independently hydrogen or C 1-6 alkyl;

[0024] R 5 , R 6 are independently hydrogen or C 1-6 Alkyl, or -NR 5 R 6 Together they represent azetidinyl, tetrahydropyrrolyl, piperidinyl, morpholinyl or 4-methylpiperazinyl.

[0025] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, Y 3 and Y 4 CH; Y 1 , Y 2 are each independently CH or N, and Y 1 With Y 2 At least one of them is N. That is, these specific embodiments of the present invention provide a compound with a structure shown in the following formula I-1:

[0026]

[0027] In Formula I-1, Y 1 , Y 2 , R 1 , R 2 Each is independently as defined above.

[0028] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, Y 1 and Y 2 N respectively.

[0029] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, Y 1 and Y 2 N, Y 3 and Y 4 That is, these specific embodiments of the present invention provide compounds with structures shown in the following formula I-2:

[0030]

[0031] In Formula I-2, R 1 , R 2 Each is independently as defined above.

[0032] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, R 1It is a 5-membered and 6-membered heteroaromatic ring group containing n heteroatoms, wherein n is an integer selected from 1 to 4, and the heteroatoms are selected from one or two of O, N, and S; the heteroaromatic ring group is optionally substituted by one or more substituents selected from halogen, -OH, -OMe, -NH2, -NHMe, -NMe2, -CN, -CONH2, and -Me.

[0033] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, R 1 is a group selected from the following:

[0034]

[0035] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof, R 2 It is hydroxy, methoxy, ethoxy, amino, methylamino, dimethylamino, cyano, methyl, ethyl, propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, cyclopropyl, hydroxymethyl, methoxymethyl, 2-hydroxyethyl, 2-methoxyethyl, 2-dimethylaminoethyl, N-morpholinyl or 4-methylpiperazinyl.

[0036] According to some specific embodiments of the present invention, in the compound of formula I or a pharmaceutically acceptable salt thereof of the present invention, the compound can be selected from:

[0037]

[0038]

[0039]

[0040] On the other hand, the present invention also provides an intermediate compound, the intermediate includes but is not limited to: any intermediate in the synthetic route for preparing the compound or its pharmaceutically acceptable salt. Preferably, the intermediate has the same basic structural unit (basically the same basic core part or basic ring) as the compound of formula I or its pharmaceutically acceptable salt, or the basic structural unit of the intermediate is included in the chemical structure of the compound of formula I or its pharmaceutically acceptable salt.

[0041] In some specific embodiments of the present invention, the intermediate compound has a structure shown in Formula II:

[0042]

[0043] In Formula II, Y 1 , Y 2 , Y 3 , Y 4With R 2 each independently as defined above;

[0044] R 3 Halogen or The halogen is preferably chlorine.

[0045] On the other hand, the present invention also provides a method for preparing a compound having a structure shown in Formula I or a pharmaceutically acceptable salt thereof, the method comprising:

[0046]

[0047] The intermediate compound shown in formula II, and R 1 -M undergoes a coupling reaction to prepare a compound having structural formula I.

[0048] in:

[0049] R 3 When it is a halogen, M is

[0050] R 3 for When M is halogen, preferably bromine;

[0051] Y 1 , Y 2 , Y 3 , Y 4 , R 1 and R 2 Each is independently as defined above.

[0052] According to a specific embodiment of the present invention, the method for preparing the compound of the structure shown in Formula I or a pharmaceutically acceptable salt thereof of the present invention also includes a process for preparing the intermediate compound.

[0053] According to some specific embodiments of the present invention, the method for preparing a compound of the structure shown in Formula I or a pharmaceutically acceptable salt thereof of the present invention includes but is not limited to: preparing a compound of Formula I or a pharmaceutically acceptable salt thereof according to any synthetic route in Preparation Example 1 to Preparation Example 34. Wherein, each substituent of the prepared compound is not limited to the scope of the specific compounds in Preparation Example 1 to Preparation Example 34, and the definition of each substituent can refer to the definition scope of each substituent of the aforementioned compound of Formula I.

[0054] On the other hand, the present invention also provides a pharmaceutical composition, which comprises: a compound of the structure shown in Formula I of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0055] The present invention also provides the use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition as an ATR kinase inhibitor. When the compound or its pharmaceutically acceptable salt or the pharmaceutical composition of the present invention is used as an ATR inhibitor, it can be for non-therapeutic purposes (such as in vitro experimental research purposes, etc.) or for therapeutic purposes. That is, the present invention provides the use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for inhibiting ATR kinase.

[0056] The present invention also provides the use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for treating a hyperproliferative disease. When the compound or its pharmaceutically acceptable salt of the present invention is used as a pharmaceutical active ingredient to treat a hyperproliferative disease, the step includes administering the compound or its pharmaceutically acceptable salt or the pharmaceutical composition to a subject in need thereof.

[0057] In the application provided by the present invention, the hyperproliferative disease includes cancer. The cancer may include, for example, melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, breast cancer, cervical cancer, ovarian cancer, prostate cancer, skin cancer, neuroblastoma, glioma, sarcoma, bone cancer, uterine cancer, endometrial cancer, head and neck tumor, multiple myeloma, B-cell lymphoma, polycythemia vera, leukemia, thyroid tumor, bladder cancer or gallbladder cancer, etc.

[0058] Definition and Description

[0059] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0060] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0061] The term "pharmaceutically acceptable salt" refers to salts of compounds of the invention, prepared from compounds of the invention having specific substituents with relatively nontoxic acids or bases. When the compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of 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 amino or magnesium salts or similar salts. When the compounds of the invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of 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, phosphate-hydrogen, 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, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid and the like; also include salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, and thus can be converted into either base or acid addition salts.

[0062] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0063] Unless otherwise specified, the key is a solid wedge. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond. and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Denotes a solid wedge bond or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key

[0064] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0065] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as 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 be substituted or not substituted, and unless otherwise specified, the type and number of the substituent can be arbitrary on the basis of chemical achievable.

[0066] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0067] When one of the variables is selected from a bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a bond, it means that the structure is actually AZ.

[0068] When one of the variables is selected as missing, it means that it does not exist. For example, when R3 in C-R3 is selected as missing, it means that the structure is actually C.

[0069] When the substituent is listed without indicating through which atom it is connected to the substituted group, such substituent can be bonded through any atom thereof. For example, a phenyl substituent can be bonded to the substituted group through any carbon atom on the benzene ring.

[0070] Unless otherwise specified, the term "alkyl" is used to refer to a straight or branched saturated hydrocarbon group, which may be monosubstituted (such as -CH2F) or polysubstituted (such as -CF3), and may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of alkyl groups include methyl (Me), ethyl (Et), propyl (such as n-propyl and isopropyl), butyl (such as n-butyl, isobutyl, s-butyl, t-butyl), pentyl (such as n-pentyl, isopentyl, neopentyl), and the like.

[0071] Unless otherwise specified, "cycloalkyl" includes any stable cyclic or polycyclic hydrocarbon radical, any carbon atom is saturated, may be monosubstituted or polysubstituted, and may be monovalent, divalent or polyvalent. Examples of such cycloalkyl radicals include, but are not limited to, cyclopropyl, norbornyl, [2.2.2]bicyclooctane, [4.4.0]bicyclodecane, and the like.

[0072] Unless otherwise specified, the term "alkenyl" means a straight or branched chain hydrocarbon group having at least one carbon-carbon double bond.2-6 "Alkenyl" means the above hydrocarbon group having 2, 3, 4, 5 or 6 carbon atoms, "C 2-4 "Alkenyl" means the above hydrocarbon group having 2, 3 or 4 carbon atoms, it should be understood that when the alkenyl contains more than one carbon-carbon double bond, the double bonds may be separated from each other or conjugated to each other. Exemplary "C 2-6 "Alkenyl" includes, but is not limited to, vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, and the like.

[0073] Unless otherwise specified, the term "alkynyl" means a straight or branched chain hydrocarbon group having at least one carbon-carbon triple bond. 2-6 "Alkynyl" means the above hydrocarbon radicals having 2, 3, 4, 5 or 6 carbon atoms, "C 2-4 "Alkynyl" means the above hydrocarbon group having 2, 3 or 4 carbon atoms, and exemplary "C 2-6 "Alkynyl" includes, but is not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, 4-methyl-1-butynyl, and the like.

[0074]

[0043] The term "halogen," by itself or as part of another substituent, means, unless otherwise stated, a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

[0075] Unless otherwise specified, the term "alkoxy" means an alkyl group attached to the rest of the molecule via an oxygen atom, wherein the alkyl group has the meaning as defined herein. 1-5 Alkoxy includes C1, C2, C3, C4 and C5 alkoxy. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S-pentoxy. The alkoxy group may be optionally substituted with one or more substituents described herein.

[0076] Unless otherwise specified, the term "amino" refers to -NH2, -NH(alkyl) or -N(alkyl)(alkyl).

[0077] Unless otherwise specified, the term "aromatic ring" means a polyunsaturated aromatic alkane monocyclic ring which may be mono- or polysubstituted.

[0078] Unless otherwise specified, the term "4-6 membered heterocycloalkyl" means a saturated monovalent monocyclic hydrocarbon ring containing 3, 4 or 5 carbon atoms and one or more selected from O, NR a A heteroatom group, wherein R a Represents hydrogen atom or C 1-6Alkyl; the "4-6 membered heterocycloalkyl" may be attached to the rest of the molecule via any carbon atom or, if present, a nitrogen atom.

[0079] Unless otherwise specified, the term "heteroaromatic ring" refers to an aromatic ring containing one to four heteroatoms selected from one or more of N, O and S.

[0080] Unless otherwise specified, the term "fused heteroaromatic ring" means a fused heterocyclic ring containing two or more aromatic groups.

[0081] Unless otherwise specified, the term "5-membered and 6-membered heteroaromatic ring" means a 5-membered and 6-membered fused heteroaromatic ring, at least one of the two fused rings contains one or more heteroatoms (including but not limited to O, S or N), and the entire group is aromatic. Non-limiting examples include benzo 5-membered heteroaromatic, 6-membered heteroaromatic ring and 5-membered heteroaromatic ring.

[0082] Unless otherwise specified, the term "6-membered and 6-membered heteroaromatic ring" means a 6-membered and 6-membered fused heteroaromatic ring, at least one of the two fused rings contains one or more heteroatoms (including but not limited to O, S or N), and the entire group is aromatic. Non-limiting examples include benzo 6-membered heteroaromatic, 6-membered heteroaromatic ring and 6-membered heteroaromatic ring.

[0083] Unless otherwise specified, the term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" refers to a salt of the compound of the present invention that retains the biological effectiveness and properties of the free acid or free base, and the free acid is obtained by reacting with a non-toxic inorganic base or organic base, or the free base is obtained by reacting with a non-toxic inorganic acid or organic acid.

[0084] Unless otherwise specified, the term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or pharmaceutically acceptable salts and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents. "Carrier" refers to a material that does not significantly irritate the organism and does not eliminate the biological activity and properties of the administered compound. "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate the administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugar, starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders and disintegrants.

[0085] The compound of the invention has good ATR enzyme inhibitory activity and tumor cell proliferation inhibitory activity, and has potential application value in treating diseases related to cell proliferation. DETAILED DESCRIPTION

[0086] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0087] The structures of the compounds were determined by nuclear magnetic resonance or mass spectrometry. Nuclear magnetic resonance was measured using a BRUKER 400M nuclear magnetic spectrometer, the solvent was deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3), the internal standard was tetramethylsilane (TMS), and the chemical shift (δ) was 10 -6 The units are given as ppm. Mass spectra were measured using a Waters ACQUITY Arc / ACQUITY QDa or Thermo U3000-ISQ EC LC / MS instrument.

[0088] Thermo U3000 high pressure liquid chromatograph was used for HPLC analysis, and Hanbang DAC-50 or Shimadzu LC-20AP preparative chromatograph was used for HPLC preparation.

[0089] Reaction monitoring uses thin layer chromatography or liquid chromatography-mass spectrometry. The developing solvent systems used in thin layer chromatography include: dichloromethane and methanol system, petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, or a small amount of triethylamine is added to adjust it. Liquid chromatography-mass spectrometry uses Waters ACQUITY Arc / ACQUITY QDa or ThermoU3000-ISQ EC liquid chromatography-mass spectrometer.

[0090] Column chromatography generally uses 200-300 mesh silica gel as a carrier. The eluent system includes: dichloromethane and methanol system, petroleum ether and ethyl acetate system, and the volume ratio of the solvent is adjusted according to the polarity of the compound, or a small amount of triethylamine is added for adjustment.

[0091] Unless otherwise specified in the following examples, the reaction temperature was room temperature (20° C. to 30° C.), and the solvents were dried and purified according to standard methods.

[0092] In conventional synthesis methods and embodiments of the present invention, the meanings of the following abbreviations are as follows:

[0093] Abbreviation meaning

[0094] Pd(dppf)Cl2 [1,1'-Bis(diphenylphosphino)ferrocene]palladium dichloride

[0095] Pd(XantPhos)Cl2 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene) Palladium dichloride

[0096] Example 1: (R)-3-methyl-4-(1'-(methylsulfonyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)morpholine (Compound 1)

[0097]

[0098] Step 1: 4-Bromo-1H-pyrrolo[2,3-b]pyridine-7-oxide

[0099] 4-Bromo-1H-pyrrolo[2,3-b]pyridine (10.0 g, 50.8 mmol) was dissolved in tetrahydrofuran (200 mL), cooled to 0°C, and m-chloroperbenzoic acid (17.5 g, 101.5 mmol) was added, and then stirred at room temperature for 4 hours to precipitate a solid. The solid was collected by filtration, dissolved, and purified by silica gel column chromatography to obtain the title compound (7.8 g, yield 72%).

[0100] LC-MS(ESI)m / z 213.0,215.0[M+H] + .

[0101] Step 2: 4-Bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine

[0102] 4-Bromo-1H-pyrrolo[2,3-b]pyridine-7-oxide (5.0 g, 23.5 mmol) was dissolved in N,N-dimethylformamide (250 mL), heated to 50°C, methanesulfonyl chloride (13.5 g, 118.0 mmol) was added, and then heated to 75°C with stirring for 1 hour. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by preparative HPLC to obtain the title compound (2.2 g, yield 41%).

[0103] LC-MS(ESI)m / z 230.9,232.9[M+H] + .

[0104] Step 3: 4-bromo-6-chloro-1-(2-(trimethylsilyl)ethoxymethyl)-1H-pyrrolo[2,3-b]pyridine

[0105] 4-Bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine (1.1 g, 4.8 mmol) was dissolved in tetrahydrofuran (35 mL), and 60% sodium hydride (400 mg, 10.0 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes. 2-(trimethylsilyl)ethoxymethyl chloride (2.5 g, 15.0 mmol) was added, and the reaction was continued to stir at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and dichloromethane (100 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. The filtrate was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (1.5 g, yield 87%).

[0106] LC-MS(ESI)m / z 361.0,363.0[M+H] + .

[0107] Step 4: (R)-4-(6-chloro-1-(2-(trimethylsilyl)ethoxymethyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine

[0108] 4-Bromo-6-chloro-1-(2-(trimethylsilyl)ethoxymethyl)-1H-pyrrolo[2,3-b]pyridine (1.0 g, 2.8 mmol) and (R)-3-methylmorpholine (280 mg, 2.8 mmol) were dissolved in 1,4-dioxane (20 mL). Degased with nitrogen and under nitrogen protection, Pd(XantPhos)Cl2 (210 mg, 0.28 mmol) and sodium tert-butoxide (270 mg, 2.8 mmol) were added, then heated to 80 ° C and stirred for 6 hours under nitrogen protection. The reaction solution was cooled to room temperature, dichloromethane (100 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (437 mg, yield 41%).

[0109] LC-MS (ESI) m / z 382.2 [M+H] + .

[0110] Step 5: (R)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine

[0111] (R)-4-(6-chloro-1-(2-(trimethylsilyl)ethoxymethyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (110 mg, 0.29 mmol) and trifluoroacetic acid (1 mL) were added to the reaction flask. The mixture was stirred at room temperature until the raw material was completely converted. The reaction solution was concentrated under reduced pressure, tetrahydrofuran (5 mL) and 1M aqueous sodium hydroxide solution (2 mL) were added, and the resulting mixture was stirred at room temperature for 30 minutes, ethyl acetate (50 mL) was added, and then washed with water. The liquids were separated and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure to obtain the crude title compound. It was not further purified and was used directly in the next step.

[0112] LC-MS (ESI) m / z 252.1 [M+H] + .

[0113] Step 6: (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine

[0114] The crude (R)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine obtained in the previous step was completely dissolved in tetrahydrofuran (6 mL), and 60% sodium hydride (48 mg, 1.2 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes, and then methanesulfonyl chloride (172 mg, 1.5 mmol) was added dropwise, and the reaction was continued to stir at room temperature for 3 hours. A small amount of water was added to quench the reaction, and ethyl acetate (40 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (76 mg, yield 80%).

[0115] LC-MS (ESI) m / z 330.1 [M+H] + .

[0116] Step 7: (R)-3-methyl-4-(1'-(methylsulfonyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)morpholine

[0117] Add (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (50 mg, 0.15 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (73.5 mg, 0.3 mmol), Pd(dppf)Cl2 (22 mg, 0.03 mmol) and sodium carbonate (47.5 mg, 0.45 mmol) to the reaction flask, and then add 1,4-dioxane (2.5 mL) and water (0.5 mL). Degas with nitrogen and heat the mixture to 100 ° C under nitrogen protection, stir and react for 3 hours. Cool the reaction solution to room temperature, add dichloromethane (20 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 1 (51 mg, yield 82%).

[0118] LC-MS (ESI) m / z 412.2 [M+H] + .

[0119] 1 H NMR (400MHz, DMSO-d6) δ11.77(s,1H),8.32(d,J=5.0Hz,1H),7.67(d,J=5.1Hz,1H),7.61(d,J =4.1Hz,1H),7.57(dd,J=3.5,2.5Hz,1H),7.31(s,1H),7.27(dd,J=3.4,1.9Hz,1H),6.98(d,J= 4.2Hz,1H),4.42-4.34(m,1H),4.03-3.95(m,1H),3.84(dd,J=11.3,2.7Hz,1H),3.79(s,3H),3 .73(d,J=11.3Hz,1H),3.69-3.59(m,2H),3.51(td,J=12.3,3.5Hz,1H),1.18(d,J=6.7Hz,3H).

[0120] 13 C NMR(100MHz,DMSO-d6)δ153.11,152.00,150.79,149.64,143.44,139.42,127.84,123.59,1 18.25,114.68,111.58,105.64,104.16,101.94,71.45,67.08,51.42,43.70,42.51,13.89.

[0121] By using a coupling reaction similar to the seventh step of Example 1, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-pyrrolo[2,3-b]pyridine is replaced with other aromatic boronic acid pinacol esters or aromatic boronic acids, and compounds 2 to 11 can be synthesized from the intermediate (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine. Aryl boronic acid pinacol esters are commercially available or prepared by Miyaura borylation reaction from aryl halides and biboronic acid pinacol esters. Aryl boronic acids are commercially available or prepared by reacting aryl halides with butyl lithium and boric acid esters.

[0122] Example 2: (R)-4-(6-(1H-indol-4-yl)-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (Compound 2)

[0123]

[0124] The reaction was carried out as described above to obtain compound 2 (54 mg, yield 87%).

[0125] LC-MS (ESI) m / z 411.2 [M+H] + .

[0126] 1 H NMR (400MHz, DMSO-d6) δ11.26(s,1H),7.57(dd,J=7.4,0.9Hz,1H),7.55(d,J=4.1Hz,1H),7.48 (d,J=8.1Hz,1H),7.44(t,J=2.8Hz,1H),7.24–7.18(m,2H),7.16(s,1H),6.94(d,J=4.2Hz,1H) ,4.36–4.28(m,1H),4.02–3.94(m,1H),3.87–3.78(m,4H),3.72(d,J=11.4Hz,1H),3.65(td,J= 11.2, 3.3Hz, 1H), 3.55 (d, J = 12.1Hz, 1H), 3.48 (td, J = 11.8, 3.4Hz, 1H), 1.17 (d, J = 6.6Hz, 3H).

[0127] Example 3: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)morpholine (Compound 3)

[0128]

[0129] The reaction was carried out as described above to obtain compound 3 (102 mg, yield 82%).

[0130] LC-MS (ESI) m / z 412.3 [M+H] + .

[0131] 1 H NMR (400MHz, DMSO-d6) δ11.76(s,1H),8.79(s,1H),8.71(s,1H),7.71(t,J=2.8Hz,1H ),7.56(d,J=4.2Hz,1H),7.32-7.27(m,1H),7.25(s,1H),6.96(d,J=4.2Hz,1H),4.41- 4.33(m,1H),3.94-4.02(m,1H),3.83(dd,J=11.6,2.8Hz,1H),3.79(s,3H),3.73(d,J =11.5Hz,1H),3.69–3.59(m,2H),3.50(td,J=12.4,3.4Hz,1H),1.19(d,J=6.6Hz,3H).

[0132] Example 4: (R)-3-methyl-4-(1'-(methylsulfonyl)-1H,1'H-[5,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)morpholine (Compound 4)

[0133]

[0134] The reaction was carried out as described above to obtain compound 4 (50 mg, yield 80%).

[0135] LC-MS (ESI) m / z 412.2 [M+H] + .

[0136] 1 H NMR (400MHz, DMSO-d6) δ11.79(s,1H),9.02(d,J=2.1Hz,1H),8.66(d,J=2.2Hz,1H) ,7.55–7.48(m,2H),7.25(s,1H),6.91(d,J=4.2Hz,1H),6.56(dd,J=3.4,1.8Hz,1H ),4.43–4.35(m,1H),3.98(dd,J=10.7,3.0Hz,1H),3.87–3.79(m,4H),3.73(d,J=1 1.6Hz,1H),3.69–3.60(m,2H),3.51(td,J=12.8,3.6Hz,1H),1.18(d,J=6.6Hz,3H).

[0137] Example 5: (R)-3-methyl-4-(1'-(methylsulfonyl)-1H,1'H-[3,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)morpholine (Compound 5)

[0138]

[0139] The reaction was carried out as described above to obtain compound 5 (53 mg, yield 85%).

[0140] LC-MS (ESI) m / z 412.2 [M+H] + .

[0141] 1 H NMR (400MHz, DMSO-d6) δ12.01(d,J=2.1Hz,1H),8.99(dd,J=7.9,1.7Hz,1H),8.37(d,J=2.8 Hz,1H),8.27(dd,J=4.7,1.7Hz,1H),7.45(d,J=4.1Hz,1H),7.21–7.14(m,2H),6.87(d,J =4.2Hz,1H),4.38–4.29(m,1H),3.98(d,J=11.3Hz,1H),3.83(dd,J=11.3,3.0Hz,1H),3. 77–3.69(m,4H),3.65(td,J=11.2,3.2Hz,1H),3.57–3.42(m,2H),1.16(d,J=6.6Hz,3H).

[0142] Example 6: (R)-3-methyl-4-(3-methyl-1'-(methylsulfonyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)morpholine (Compound 6)

[0143]

[0144] The reaction was carried out as described above to obtain compound 6 (33 mg, yield 34%).

[0145] LC-MS (ESI) m / z 426.2 [M+H] + .

[0146] 1H NMR (400MHz, DMSO-d6) δ11.47(s,1H),8.24(d,J=4.8Hz,1H),7.58(d,J=4.2Hz,1H) ,7.30–7.23(m,1H),7.08(d,J=4.8Hz,1H),6.98(d,J=4.2Hz,1H),6.82(s,1H),4.37 –4.29(m,1H),3.97–3.89(m,1H),3.80(dd,J=11.3,2.8Hz,1H),3.74–3.67(m,4H),3 .61(td,J=11.3,3.0Hz,1H),3.55–3.41(m,2H),1.94(s,3H),1.16(d,J=6.6Hz,3H).

[0147] Example 7: (R)-4-(6-(6-fluoro-1H-indol-4-yl)-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (Compound 7)

[0148]

[0149] The reaction was carried out as described above to obtain compound 7 (56 mg, yield 86%).

[0150] LC-MS (ESI) m / z 429.2 [M+H] + .

[0151] 1 H NMR (400MHz, DMSO-d6) δ11.32(s,1H),7.57(d,J=4.1Hz,1H),7.47(dd,J=11.2,2.3Hz,1H),7 .45–7.43(m,1H),7.26(dd,J=9.5,2.2Hz,1H),7.23–7.20(m,1H),7.18(s,1H),6.95(d,J=4.2 Hz,1H),4.40–4.32(m,1H),4.02–3.94(m,1H),3.83(dd,J=11.4,2.7Hz,1H),3.79(s,3H),3. 72(d,J=11.2Hz,1H),3.69–3.57(m,2H),3.49(td,J=12.4,3.5Hz,1H),1.18(d,J=6.7Hz,3H).

[0152] Example 8: (R)-4-(6-(7-fluoro-1H-indol-4-yl)-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (Compound 8)

[0153]

[0154] The reaction was carried out as described above to obtain compound 8 (35 mg, yield 54%).

[0155] LC-MS (ESI) m / z 429.2 [M+H] + .

[0156] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),7.59–7.53(m,2H),7.51(t,J=2.7Hz,1H),7.33–7.29(m ,1H),7.13(s,1H),7.04(dd,J=11.0,8.2Hz,1H),6.93(d,J=4.2Hz,1H),4.37–4.29(m,1H),4.0 2–3.94(m,1H),3.83(dd,J=11.3,2.9Hz,1H),3.79(s,3H),3.72(d,J=10.6Hz,1H),3.65(td,J= 11.3, 2.9Hz, 1H), 3.56 (d, J = 11.6Hz, 1H), 3.48 (td, J = 12.2, 3.5Hz, 1H), 1.17 (d, J = 6.6Hz, 3H).

[0157] Example 9: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)morpholine (Compound 9)

[0158]

[0159] The reaction was carried out as described above to obtain compound 9 (45 mg, yield 82%).

[0160] LC-MS (ESI) m / z 362.2 [M+H] + .

[0161] 1H NMR (400MHz, DMSO-d6) δ12.98(s,1H),7.82(s,1H),7.51(d,J=4.1Hz,1H),7.31(s,1H),6.90(d,J=4.2Hz,1H),6.83(s,1H),4.34–4.22( m,1H),3.98(d,J=10.8Hz,1H),3.93–3.77(m,4H),3.71(d,J=11.2Hz,1H),3.67–3.57(m,1H),3.50–3.39(m,2H),1.14(d,J=6.5Hz,3H).

[0162] Example 10: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)morpholine (Compound 10)

[0163]

[0164] The reaction was carried out as described above to obtain compound 10 (43 mg, yield 78%).

[0165] LC-MS (ESI) m / z 362.2 [M+H] + .

[0166] 1 H NMR (400MHz, DMSO-d6) δ13.03(s,1H),8.35(s,1H),8.08(s,1H),7.44(d,J=4.1 Hz,1H),6.98(s,1H),6.84(d,J=4.1Hz,1H),4.35–4.27(m,1H),3.96(d,J=13.1 Hz,1H),3.85–3.77(m,4H),3.71(d,J=11.3Hz,1H),3.62(td,J=11.5,3.3Hz,1H ), 3.51 (d, J = 12.1Hz, 1H), 3.45 (dd, J = 11.4, 3.4Hz, 1H), 1.13 (d, J = 6.6Hz, 3H).

[0167] Example 11: (R)-4-(6-(1H-benzo[d]imidazol-2-yl)-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (Compound 11)

[0168]

[0169] The reaction was carried out as described above to obtain compound 11 (23 mg, yield 40%).

[0170] LC-MS (ESI) m / z 412.2 [M+H] + .

[0171] 1 H NMR (400MHz, DMSO-d6) δ12.68(s,1H),7.73–7.68(m,2H),7.66–7.61(m,2H),7.30–7.19(m,2H),6.99(d,J=4.2Hz,1H),4.39–4.31(m,1H),4.0 6–3.96(m,4H),3.84(dd,J=11.5,2.5Hz,1H),3.74(d,J=11.2Hz,1H),3.65(td,J=11.0,3.8Hz,1H),3.59–3.50(m,2H),1.18(d,J=6.6Hz,3H).

[0172] Example 12: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-morpholine (Compound 12)

[0173]

[0174] Step 1: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)morpholine

[0175] Add (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (200 mg, 0.61 mmol), biboronic acid pinacol ester (230 mg, 0.91 mmol), Pd(dppf)Cl2 (88 mg, 0.12 mmol) and potassium acetate (101 mg, 1.03 mmol) to the reaction bottle, and then add 1,4-dioxane (4 mL). Degas with nitrogen and heat the above mixture to 110 ° C under nitrogen protection, and stir the reaction for 1.5 hours. The reaction solution was cooled to room temperature, ethyl acetate (20 mL) was added, and the resulting mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain the crude product of the title compound (300 mg). It was not further purified and used directly in the next step.

[0176] Step 2: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-morpholine

[0177] Add (R)-3-methyl-4-(1-(methylsulfonyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)morpholine (200 mg crude product, calculated as 0.40 mmol), 4-bromo-7H-pyrrolo[2,3-d]pyrimidine (48 mg, 0.24 mmol), Pd(dppf)Cl2 (51 mg, 0.07 mmol) and sodium carbonate (153 mg, 1.44 mmol) to the reaction flask, and then add 1,4-dioxane (5 mL) and water (1 mL). Degas with nitrogen and heat the mixture to 85°C under nitrogen protection, and stir to react for 1 hour. Cool the reaction solution to room temperature, add dichloromethane (25 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 12 (40 mg, yield 40%).

[0178] LC-MS (ESI) m / z 413.2 [M+H] + .

[0179] 1 H NMR (400MHz, DMSO-d6) δ12.19(s,1H),8.85(s,1H),8.01(s,1H),7.72–7.63(m,3H),7.02(d,J=4.2Hz,1H),4.40–4.32(m,1H),4.02(d,J =11.0Hz,1H),3.85(dd,J=11.5,2.8Hz,1H),3.79–3.70(m,4H),3.65(td,J=10.7,4.0Hz,1H),3.59–3.50(m,2H),1.18(d,J=6.6Hz,3H).

[0180] Example 13: (R)-4-(6-(3H-imidazo[4,5-b]pyridin-7-yl)-1-(methylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine (Compound 13)

[0181]

[0182] The reaction was carried out as described above (similar to the second step of Example 12) to obtain compound 13 (20 mg, yield 35%).

[0183] LC-MS (ESI) m / z 413.1 [M+H] + .

[0184] 1H NMR (400MHz, CDCl3) δ12.00(s,1H),8.66(d,J=5.1Hz,1H),8.41(s,1H),7.65(d,J=5.2Hz,1 H),7.51(d,J=4.1Hz,1H),7.29(s,1H),6.74(d,J=4.1Hz,1H),4.31–4.23(m,1H),4.15–4.0 8(m,1H),3.97(dd,J=11.4,3.0Hz,1H),3.85(d,J=11.0Hz,1H),3.80(td,J=11.4,2.9Hz,1H ),3.68(td,J=11.7,3.5Hz,1H),3.53(d,J=12.1Hz,1H),3.40(s,3H),1.34(d,J=6.7Hz,3H).

[0185] Example 14: (R)-4-(1'-(isopropylsulfonyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)-3-methylmorpholine (Compound 14)

[0186]

[0187] By adopting a reaction similar to the sixth and seventh steps of Example 1, the methanesulfonyl chloride in the sixth step of Example 1 was replaced with isopropylsulfonyl chloride, and the intermediate (R)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine was reacted in two steps as shown above to obtain compound 14 (111 mg, two-step yields were 92% and 45%, respectively).

[0188] LC-MS (ESI) m / z 440.2 [M+H] + .

[0189] 1H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.31(d,J=5.0Hz,1H),7.68(d,J=5.1Hz,1H),7.61–7.56(m ,2H),7.36(dd,J=3.5,2.0Hz,1H),7.33(s,1H),7.00(d,J=4.2Hz,1H),4.45–4.35(m,2H),3.99(d d,J=11.7,2.5Hz,1H),3.83(dd,J=11.4,2.8Hz,1H),3.73(d,J=11.3Hz,1H),3.69–3.60(m,2H),3 .51(td,J=12.7,3.4Hz,1H),1.29(d,J=6.9Hz,3H),1.23(d,J=6.8Hz,3H),1.19(d,J=6.6Hz,3H).

[0190] Example 15: (R)-4-(1'-(cyclopropylsulfonyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-4'-yl)-3-methylmorpholine (Compound 15)

[0191]

[0192] By adopting the reaction similar to the sixth and seventh steps of Example 1, the methanesulfonyl chloride in the sixth step of Example 1 was replaced by cyclopropylsulfonyl chloride, and the intermediate (R)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine was reacted in two steps as shown above to obtain compound 15 (109 mg, two-step yields were 90% and 50%, respectively).

[0193] LC-MS (ESI) m / z 438.2 [M+H] + .

[0194] 1H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.32(d,J=5.0Hz,1H),7.68(d,J=5.1Hz,1H),7.60–7.55(m ,2H),7.35(dd,J=3.5,1.9Hz,1H),7.32(s,1H),6.96(d,J=4.2Hz,1H),4.41–4.33(m,1H),4.02–3 .96(m,1H),3.84(dd,J=11.4,2.8Hz,1H),3.73(d,J=11.1Hz,1H),3.70–3.59(m,2H),3.51(td,J= 12.5,3.7Hz,1H),3.47–3.39(m,1H),1.43–1.29(m,2H),1.19(d,J=6.6Hz,3H),1.14–1.06(m,2H).

[0195] Example 16: (R)-N,N-dimethyl-4'-(3-methylmorpholinyl)-1H,1'H-[4,6'-dipyrrolo[2,3-b]pyridine]-1'-sulfonamide (Compound 16)

[0196]

[0197] A reaction similar to the sixth and seventh steps of Example 1 was adopted, and the methanesulfonyl chloride in the sixth step of Example 1 was replaced with dimethylaminosulfonyl chloride. The intermediate (R)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-4-yl)-3-methylmorpholine was reacted in two steps as shown above to obtain compound 16 (148 mg, two-step yields were 91% and 60%, respectively).

[0198] LC-MS (ESI) m / z 441.2 [M+H] + .

[0199] 1H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.31(d,J=5.0Hz,1H),7.61(d,J=5.1Hz,1H),7. 59–7.55(m,2H),7.28(dd,J=3.4,1.9Hz,1H),7.25(s,1H),6.91(d,J=4.2Hz,1H),4.39 –4.31(m,1H),4.02–3.95(m,1H),3.84(dd,J=11.4,2.7Hz,1H),3.72(d,J=11.2Hz,1H) ,3.69–3.56(m,2H),3.50(td,J=12.0,3.4Hz,1H),2.89(s,6H),1.18(d,J=6.6Hz,3H).

[0200] Example 17: (R)-3-methyl-4-(7-(methylsulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (Compound 17)

[0201]

[0202] Step 1: (R)-4-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine

[0203] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (1.00 g, 5.3 mmol) and (R)-3-methylmorpholine (2.70 g, 26.7 mmol) were dissolved in 1,4-dioxane (25 mL). Triethylamine (3.80 g, 37.6 mmol) was slowly added, and the mixture was heated to 90 °C and stirred for 8 hours. The reaction solution was cooled to room temperature, dichloromethane (100 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain the title compound (1.02 g, yield 76%).

[0204] LC-MS (ESI) m / z 253.1 [M+H] + .

[0205] Step 2: (R)-4-(2-chloro-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine

[0206] (R)-4-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (152 mg, 0.6 mmol) was dissolved in tetrahydrofuran (7.5 mL), and 60% sodium hydride (48 mg, 1.2 mmol) was slowly added at room temperature. The reaction was stirred for 5 minutes, and then methanesulfonyl chloride (137 mg, 1.2 mmol) was added dropwise. The reaction was continued to stir at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and dichloromethane (100 mL) was added, and then washed with water. The liquid was separated, and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (149 mg, yield 75%).

[0207] LC-MS (ESI) m / z 331.1 [M+H] + .

[0208] Step 3: (R)-3-methyl-4-(7-(methylsulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine

[0209] Add (R)-4-(2-chloro-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (180 mg, 0.54 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (264 mg, 1.08 mmol), Pd(dppf)Cl2 (80 mg, 0.11 mmol) and potassium carbonate (299 mg, 2.16 mmol) to the reaction flask, then add 1,4-dioxane (17.5 mL) and water (3.5 mL). Degas with nitrogen and heat the mixture to 90°C under nitrogen protection, stir and react for 1 hour. Cool the reaction solution to room temperature, add dichloromethane (100 mL), and then wash with water. The organic phase was separated and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 17 (130 mg, yield 58%).

[0210] LC-MS (ESI) m / z 413.1 [M+H] + .

[0211] 1H NMR (400MHz, DMSO-d6) δ11.79(s,1H),8.35(d,J=5.0Hz,1H),8.04(d,J=5.0Hz ,1H),7.63–7.57(m,2H),7.48(dd,J=3.2,1.9Hz,1H),7.02(d,J=4.1Hz,1H),4. 92–4.80(m,1H),4.62–4.50(m,1H),4.04(dd,J=11.1,3.0Hz,1H),3.85–3.71( m,5H),3.60(td,J=12.2,3.2Hz,1H),3.55–3.45(m,1H),1.36(d,J=6.8Hz,3H).

[0212] 13 C NMR (100MHz, DMSO-d6) δ158.60,157.15,153.14,151.17,143.19,137.50,128.22,123. 07,118.76,115.52,105.90,104.00,102.74,71.17,67.13,48.81,48.79,42.64,15.25.

[0213] By using a coupling reaction similar to the third step of Example 17, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine was replaced with other aromatic boronic acid pinacol esters or aromatic boronic acids, and compounds 18 to 20 and compound 27 can be synthesized from the intermediate (R)-4-(2-chloro-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine.

[0214] Example 18: (R)-4-(2-(1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 18)

[0215]

[0216] The reaction was carried out as described above to obtain compound 18 (52 mg, yield 42%).

[0217] LC-MS (ESI) m / z 412.2 [M+H] + .

[0218] 1H NMR (400MHz, DMSO-d6) δ11.26(s,1H),8.15(d,J=7.5Hz,1H),7.62–7.42(m,4H),7.21(t,J=7.7Hz,1H),6.97(d,J=4.1Hz,1H),4.92– 4.77(m,1H),4.61–4.46(m,1H),4.09–3.99(m,1H),3.85–3.70(m,5H),3.65–3.55(m,1H),3.53–3.42(m,1H),1.35(d,J=6.7Hz,3H).

[0219] Example 19: (R)-3-methyl-4-(7-(methylsulfonyl)-2-(1H-pyrrolo[2,3-c]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (Compound 19)

[0220]

[0221] The reaction was carried out as described above to obtain compound 19 (81 mg, yield 65%).

[0222] LC-MS (ESI) m / z 413.2 [M+H] + .

[0223] 1 H NMR (400MHz, DMSO-d6) δ11.79(s,1H),9.17(s,1H),8.83(s,1H),7.74(t,J=2.6Hz,1H),7.56–7.51(m,2H),6.99(d,J=4.1Hz,1H),4.91–4.80(m, 1H),4.61–4.48(m,1H),4.04(dd,J=10.8,2.9Hz,1H),3.85–3.71(m,5H) ,3.60(td,J=11.8,2.8Hz,1H),3.55–3.45(m,1H),1.36(d,J=6.7Hz,3H).

[0224] Example 20: (R)-4-(2-(6-Fluoro-1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 20)

[0225]

[0226] The reaction was carried out as described above to obtain compound 20 (35 mg, yield 34%).

[0227] LC-MS (ESI) m / z 430.2 [M+H] + .

[0228] 1 H NMR (400MHz, DMSO-d6) δ11.33 (s, 1H), 7.92 (dd, J = 11.5, 2.5Hz, 1H), 7.59 (brs, 1H),7.55(d,J=4.0Hz,1H),7.48(t,J=2.8Hz,1H),7.32(dd,J=9.2,2.5Hz,1H),6 .99(d,J=4.2Hz,1H),4.90–4.76(m,1H),4.59–4.46(m,1H),4.08–4.00(m,1H), 3.85–3.71(m,5H),3.64–3.55(m,1H),3.53–3.43(m,1H),1.35(d,J=6.8Hz,3H).

[0229] Example 21: (R)-2-((4-(3-methylmorpholinyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)sulfonyl)ethanol (Compound 21)

[0230]

[0231] Step 1: (R)-4-(2-chloro-7-((2-methoxyethyl)sulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine

[0232] (R)-4-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (200 mg, 0.79 mmol) was dissolved in tetrahydrofuran (5 mL), 60% sodium hydride (63 mg, 1.58 mmol) was slowly added at room temperature, the reaction was stirred for 5 minutes, and 2-methoxyethylsulfonyl chloride (251 mg, 1.58 mmol) was added dropwise. The reaction was continued to be stirred at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and ethyl acetate (50 mL) was added, and then washed with water. The liquid was separated, and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (273 mg, yield 92%).

[0233] LC-MS (ESI) m / z 375.1 [M+H] + .

[0234] Step 2: (R)-2-((2-chloro-4-(3-methylmorpholinyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)sulfonyl)ethanol

[0235] (R)-4-(2-chloro-7-((2-methoxyethyl)sulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (270 mg, 0.72 mmol) was dissolved in dichloromethane (20 mL), cooled to 0°C, added with boron tribromide (1.21 g, 4.83 mmol), and continued to stir at room temperature for 1 hour. Water (20 mL) was added to quench the reaction, and the dichloromethane was removed by concentration under reduced pressure, and then extracted with ethyl acetate (50 mL), separated, and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (223 mg, yield 86%).

[0236] LC-MS (ESI) m / z 361.1 [M+H] + .

[0237] Step 3: (R)-2-((4-(3-methylmorpholinyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)sulfonyl)ethanol

[0238] Add (R)-2-((2-chloro-4-(3-methylmorpholinyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)sulfonyl)ethanol (100 mg, 0.28 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (135 mg, 0.55 mmol), Pd(dppf)Cl2 (40 mg, 0.055 mmol) and potassium acetate (27 mg, 0.28 mmol) to the reaction bottle, and then add 1,4-dioxane (4 mL). Degas with nitrogen and heat the mixture to 85 ° C under nitrogen protection, stir and react for 4 hours. Cool the reaction solution to room temperature, add ethyl acetate (50 mL), and then wash with water (20 mL). Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 21 (49 mg, yield 40%).

[0239] LC-MS (ESI) m / z 443.2 [M+H] + .

[0240] 1H NMR (400MHz, DMSO-d6) δ11.77(s,1H),8.34(d,J=5.0Hz,1H),8.04(d,J=5.0Hz,1H),7.60(t,J=3 .0Hz,1H),7.54(d,J=4.1Hz,2H),7.51(dd,J=3.4,2.0Hz,1H),6.99(d,J=4.1Hz,1H),4.94–4.80( m,2H),4.62–4.48(m,1H),4.13(t,J=5.9Hz,2H),4.04(dd,J=11.3,3.5Hz,1H),3.81(d,J=11.5Hz ,1H),3.78–3.68(m,3H),3.60(td,J=11.6,2.7Hz,1H),3.55–3.45(m,1H),1.36(d,J=6.7Hz,3H).

[0241] Example 22: (R)-3-methyl-4-(9-(methylsulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-9H-purin-6-yl)morpholine (Compound 22)

[0242]

[0243] Step 1: (R)-4-(2-chloro-9H-purin-6-yl)-3-methylmorpholine

[0244] 2,6-Dichloro-9H-purine (1.0 g, 5.3 mmol) was dissolved in N,N-dimethylformamide (15 mL), (R)-3-methylmorpholine (2.1 g, 21 mmol) and triethylamine (5.4 g, 53 mmol) were slowly added, and then heated to 120°C and stirred for 1 hour. The reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the title compound (1.2 g, yield 89%).

[0245] LC-MS (ESI) m / z 254.1 [M+H] + .

[0246] Step 2: (R)-4-(2-chloro-9-(methylsulfonyl)-9H-purin-6-yl)-3-methylmorpholine

[0247] (R)-4-(2-chloro-9H-purine-6-yl)-3-methylmorpholine (800 mg, 3.15 mmol) was dissolved in tetrahydrofuran (15 mL), and 60% sodium hydride (505 mg, 12.6 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes, and then methanesulfonyl chloride (1.44 g, 12.6 mmol) was added dropwise. The reaction was continued to stir at room temperature for 3 hours. A small amount of water was added to quench the reaction, and ethyl acetate (50 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (785 mg, yield 75%).

[0248] LC-MS (ESI) m / z 332.1 [M+H] + .

[0249] Step 3: (R)-3-methyl-4-(9-(methylsulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-9H-purin-6-yl)morpholine

[0250] Add (R)-4-(2-chloro-9-(methylsulfonyl)-9H-purin-6-yl)-3-methylmorpholine (100 mg, 0.30 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (183 mg, 0.75 mmol), Pd(dppf)Cl2 (44 mg, 0.060 mmol) and potassium acetate (118 mg, 1.20 mmol) to the reaction flask, and then add 1,4-dioxane (10 mL) and water (0.25 mL). Degas with nitrogen and heat the mixture to 85 ° C under nitrogen protection, stir and react for 1 hour. Cool the reaction solution to room temperature, add ethyl acetate (20 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 22 (31 mg, yield 25%).

[0251] LC-MS (ESI) m / z 414.2 [M+H] + .

[0252] 1H NMR (400MHz, DMSO-d6) δ11.83(s,1H),8.55(s,1H),8.36(d,J=5.0Hz,1H),8.04( d,J=5.0Hz,1H),7.63(t,J=3.0Hz,1H),7.39(dd,J=3.4,2.0Hz,1H),5.56–5.38( br,1H),5.18–5.06(br,1H),4.05(d,J=8.6Hz,1H),3.88(s,3H),3.84(d,J=11.2 Hz,1H),3.75(dd,J=11.5,3.0Hz,1H),3.65–3.47(m,2H),1.40(d,J=6.8Hz,3H).

[0253] Example 23: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine (Compound 23)

[0254]

[0255] Step 1: (R)-4-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylmorpholine

[0256] 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine (0.40 g, 2.1 mmol) was dissolved in 1,4-dioxane (2 mL), (R)-3-methylmorpholine (0.86 g, 8.5 mmol) and triethylamine (2.14 g, 21.1 mmol) were slowly added, and the mixture was heated to 120°C and stirred for 2 hours. The reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the title compound (0.50 g, yield 93%).

[0257] LC-MS (ESI) m / z 254.1 [M+H] + .

[0258] Step 2: (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylmorpholine

[0259] (R)-4-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylmorpholine (650 mg, 2.56 mmol) was dissolved in tetrahydrofuran (4 mL), and 60% sodium hydride (205 mg, 5.13 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes, and then methanesulfonyl chloride (587 mg, 5.12 mmol) was added dropwise. The reaction was continued to stir at room temperature for 1 hour. A small amount of water was added to quench the reaction, and ethyl acetate (30 mL) was added, and then washed with water. The liquid was separated, and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (765 mg, yield 90%).

[0260] LC-MS (ESI) m / z 332.1 [M+H] + .

[0261] Step 3: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)morpholine

[0262] Add (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-methylmorpholine (100 mg, 0.30 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (147 mg, 0.60 mmol), Pd(dppf)Cl2 (44 mg, 0.060 mmol) and potassium fluoride (70 mg, 1.20 mmol) to the reaction flask, and then add 1,4-dioxane (5 mL). Degas with nitrogen and heat the mixture to 90 ° C under nitrogen protection, stir and react for 1 hour. Cool the reaction solution to room temperature, add ethyl acetate (20 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 23 (19 mg, yield 15%).

[0263] LC-MS (ESI) m / z 414.1 [M+H] + .

[0264] 1H NMR (400MHz, DMSO-d6) δ11.84(s,1H),8.70(s,1H),8.37(d,J=5.0Hz,1H),8.10(d,J=5.0Hz,1H),7.66–7.62(m,1H),7.55–7.51(m,1H),4.72 –7.42(br,2H),4.07(d,J=8.4Hz,1H),3.84(d,J=12.0Hz,1H),3.77(d,J=12.0Hz,1H),3.70(s,3H),3.67–3.48(m,2H),1.40(d,J=6.7Hz,3H).

[0265] Example 24: (R)-3-methyl-4-(3-(methylsulfonyl)-5-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)morpholine (Compound 24)

[0266]

[0267] Step 1: (R)-4-(5-chloro-3H-imidazo[4,5-b]pyridin-7-yl)-3-methylmorpholine

[0268] Dissolve 5,7-dichloro-3H-imidazo[4,5-b]pyridine (500 mg, 2.66 mmol) in N,N-diisopropylethylamine (9 mL, 51.7 mmol), add (R)-3-methylmorpholine (3 mL, 26.4 mmol) at room temperature, and heat to 145 ° C and stir for 8 days. Cool the reaction solution to room temperature, add ethyl acetate (50 mL), and then wash with water. Separate the liquid, and dry the organic phase with anhydrous sodium sulfate. Filter, concentrate the filtrate under reduced pressure, and purify it by silica gel column chromatography to obtain the title compound (605 mg, yield 90%).

[0269] LC-MS (ESI) m / z 253.1 [M+H] + .

[0270] Step 2: (R)-4-(5-chloro-3-(methylsulfonyl)-3H-imidazo[4,5-b]pyridin-7-yl)-3-methylmorpholine

[0271] (R)-4-(5-chloro-3H-imidazo[4,5-b]pyridin-7-yl)-3-methylmorpholine (350 mg, 1.39 mmol) was dissolved in tetrahydrofuran (20 mL), and 60% sodium hydride (180 mg, 4.50 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes, and then methanesulfonyl chloride (518 mg, 4.52 mmol) was added dropwise. The reaction was continued to be stirred at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and dichloromethane (100 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (357 mg, yield 78%).

[0272] LC-MS (ESI) m / z 331.1 [M+H] + .

[0273] Step 3: (R)-3-methyl-4-(3-(methylsulfonyl)-5-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)morpholine

[0274] Add (R)-4-(5-chloro-3-(methylsulfonyl)-3H-imidazo[4,5-b]pyridin-7-yl)-3-methylmorpholine (20 mg, 0.06 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (30 mg, 0.12 mmol), Pd(dppf)Cl2 (9 mg, 0.012 mmol) and potassium fluoride (20 mg, 0.344 mmol) to the reaction flask, and then add 1,4-dioxane (2 mL) and water (0.1 mL). Degas with nitrogen and heat the mixture to 85 ° C under nitrogen protection, stir and react for 2 hours. Cool the reaction solution to room temperature, add dichloromethane (50 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 24 (11 mg, yield 44%).

[0275] LC-MS (ESI) m / z 413.1 [M+H] + .

[0276] 1H NMR (400MHz, DMSO-d6) δ11.79(s,1H),8.51(s,1H),8.32(d,J=5.0Hz,1H),7.66(d,J=5.0 Hz,1H),7.58(dd,J=3.4,2.5Hz,1H),7.30(s,1H),7.17(dd,J=3.4,1.9Hz,1H),5.25–5.1 4(m,1H),4.48–4.33(m,1H),4.02(dd,J=11.5,3.5Hz,1H),3.87(s,3H),3.79(d,J=2.2Hz ,2H),3.64(td,J=11.7,2.7Hz,1H),3.45(td,J=12.7,3.6Hz,1H),1.26(d,J=6.7Hz,3H).

[0277] Example 25: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-imidazo[4,5-c]pyridin-4-yl)morpholine (Compound 25)

[0278]

[0279] (R)-4-(6-Chloro-1H-imidazo[4,5-c]pyridin-4-yl)-3-methylmorpholine was prepared according to the synthetic route described in the literature (ACSMed. Chem. Lett. 2015, 6, 42-46).

[0280] Step 1: (R)-4-(6-chloro-1-(methylsulfonyl)-1H-imidazo[4,5-c]pyridin-4-yl)-3-methylmorpholine

[0281] (R)-4-(6-chloro-1H-imidazo[4,5-c]pyridin-4-yl)-3-methylmorpholine (150 mg, 0.59 mmol) was dissolved in tetrahydrofuran (10 mL), and 60% sodium hydride (48 mg, 1.20 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes. Then, methanesulfonyl chloride (136 mg, 1.19 mmol) was added dropwise, and the reaction was continued to be stirred at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and ethyl acetate (200 mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. The filtrate was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (150 mg, yield 76%).

[0282] LC-MS (ESI) m / z 331.1 [M+H] + .

[0283] Step 2: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-c]pyridin-4-yl)-1H-imidazo[4,5-c]pyridin-4-yl)morpholine

[0284] To the reaction flask, (R)-4-(6-chloro-1-(methylsulfonyl)-1H-imidazo[4,5-c]pyridin-4-yl)-3-methylmorpholine (50 mg, 0.15 mmol), (1H-pyrrolo[2,3-c]pyridin-4-yl)boric acid (49 mg, 0.30 mmol), Pd(dppf)Cl2 (11 mg, 0.015 mmol) and sodium carbonate (32 mg, 0.30 mmol) were added, and 1,4-dioxane (1.5 mL) and water (0.5 mL) were added. Degassing with nitrogen and under nitrogen protection, the above mixture was heated to 80 ° C and stirred for 2 hours. The reaction solution was cooled to room temperature, ethyl acetate (50 mL) was added, and then washed with water. The liquid was separated and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by preparative HPLC to obtain compound 25 (20 mg, yield 32%).

[0285] LC-MS (ESI) m / z 413.1 [M+H] + .

[0286] 1 H NMR(400MHz,DMSO-d6)δ11.79(s,1H),8.78(s,1H),8.66(s,1H),8.49(s,1 H),7.73(t,J=2.7Hz,1H),7.64(s,1H),6.94–6.89(m,1H),5.48–5.38(m,1 H),4.89(d,J=13.4Hz,1H),4.01(dd,J=11.1,2.7Hz,1H),3.83–3.72(m,5H ),3.60(td,J=11.7,2.6Hz,1H),3.47–3.38(m,1H),1.31(d,J=6.7Hz,3H).

[0287] Example 26: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrrolo[3,2-c]pyridin-4-yl)morpholine (Compound 26)

[0288]

[0289] Step 1: (R)-4-(6-chloro-1H-pyrrolo[3,2-c]pyridin-4-yl)-3-methylmorpholine

[0290] 4,6-Dichloro-1H-pyrrolo[3,2-c]pyridine (500mg, 2.67mmol) was dissolved in N,N-diisopropylethylamine (9mL, 51.7mmol), (R)-3-methylmorpholine (3mL, 26.4mmol) was added at room temperature, and then heated to 145°C and stirred for 8 days. The reaction solution was cooled to room temperature, ethyl acetate (50mL) was added, and then washed with water. The organic phase was separated and dried over anhydrous sodium sulfate. The filtrate was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (603mg, yield 90%).

[0291] LC-MS (ESI) m / z 252.1 [M+H] + .

[0292] Step 2: (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[3,2-c]pyridin-4-yl)-3-methylmorpholine

[0293] (R)-4-(6-chloro-1H-pyrrolo[3,2-c]pyridin-4-yl)-3-methylmorpholine (550 mg, 2.19 mmol) was dissolved in tetrahydrofuran (20 mL), and 60% sodium hydride (367 mg, 9.18 mmol) was slowly added at room temperature, and the reaction was stirred for 5 minutes, and then methanesulfonyl chloride (740 mg, 6.46 mmol) was added dropwise. The reaction was continued to be stirred at room temperature until the raw material was completely converted. A small amount of water was added to quench the reaction, and dichloromethane (100 mL) was added, and then washed with water. The liquid was separated, and the organic phase was dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (591 mg, yield 82%).

[0294] LC-MS (ESI) m / z 330.1 [M+H] + .

[0295] Step 3: (R)-3-methyl-4-(1-(methylsulfonyl)-6-(1H-pyrrolo[2,3-b]pyridin-4-yl)-1H-pyrrolo[3,2-c]pyridin-4-yl)morpholine

[0296] Add (R)-4-(6-chloro-1-(methylsulfonyl)-1H-pyrrolo[3,2-c]pyridin-4-yl)-3-methylmorpholine (20 mg, 0.06 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine (30 mg, 0.12 mmol), Pd(dppf)Cl2 (9 mg, 0.012 mmol) and sodium carbonate (20 mg, 0.19 mmol) to the reaction flask, and then add 1,4-dioxane (1 mL) and water (0.2 mL). Degas with nitrogen and heat the mixture to 100 ° C under nitrogen protection, stir and react for 2 hours. Cool the reaction solution to room temperature, add dichloromethane (20 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 26 (10 mg, yield 40%).

[0297] LC-MS (ESI) m / z 412.2 [M+H] + .

[0298] 1 H NMR(400MHz,DMSO-d6)δ11.79(s,1H),8.31(d,J=5.0Hz,1H),7.93(s,1H),7.64–7.61(m,2H),7 .59(t,J=3.0Hz,1H),7.08(d,J=3.8Hz,1H),6.90(dd,J=3.5,1.9Hz,1H),4.66–4.58(m,1H),4. 08(d,J=12.8Hz,1H),4.02–3.96(m,1H),3.80(dd,J=11.4,2.9Hz,1H),3.73(d,J=10.8Hz,1H), 3.63(td,J=11.3,2.6Hz,1H),3.58(s,3H),3.49(td,J=12.5,3.4Hz,1H),1.26(d,J=6.7Hz,3H).

[0299] Example 27: (R)-4-(2-(1H-benzo[d]imidazol-2-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 27)

[0300]

[0301] The reaction was carried out as described above (similar to the third step of Example 17) to obtain compound 27 (30 mg, yield 35%).

[0302] LC-MS (ESI) m / z 413.2 [M+H] + .

[0303] 1 H NMR(400MHz, DMSO-d6)δ9.19(s,1H),8.65(d,J=8.0Hz,1H),7.78(d,J=7.9Hz,1H),7.52 (d,J=4.1Hz,1H),7.45–7.39(m,1H),7.38–7.32(m,1H),7.03(d,J=4.1Hz,1H),4.91–4.7 5(br,1H),4.65–4.49(br,1H),4.02(dd,J=11.2,3.0Hz,1H),3.80(d,J=11.5Hz,1H),3. 77–3.70(m,4H),3.59(td,J=11.6,2.5Hz,1H),3.54–3.45(m,1H),1.37(d,J=6.8Hz,3H).

[0304] Example 28: (R)-N-methyl-1-(4-(3-methylmorpholinyl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (Compound 28)

[0305]

[0306] Add (R)-4-(2-chloro-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (100 mg, 0.30 mmol), N-methyl-1H-benzo[d]imidazole-2-amine (89 mg, 0.60 mmol), tris(dibenzylideneacetone)dipalladium (28 mg, 0.031 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (29 mg, 0.061 mmol) and cesium carbonate (296 mg, 0.91 mmol) to the reaction bottle, and then add 1,4-dioxane (2 mL). Degas with nitrogen and heat the mixture to 85°C under nitrogen protection, stir and react for 1 hour. Cool the reaction solution to room temperature, add dichloromethane (80 mL), and wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 28 (28 mg, yield 21%).

[0307] LC-MS (ESI) m / z 442.2 [M+H] + .

[0308] 1H NMR (400MHz, DMSO-d6) δ8.56(q,J=4.5Hz,1H),8.24(d,J=7.9Hz,1H),7.51(d,J=4. 1Hz,1H),7.28(d,J=7.7Hz,1H),7.14–7.08(m,1H),7.05(d,J=4.1Hz,1H),7.04–6. 98(m,1H),4.78–4.68(m,1H),4.41–4.31(m,1H),4.10–4.01(m,1H),3.86–3.72(m, 2H),3.65(s,3H),3.63–3.51(m,2H),3.08(d,J=4.8Hz,3H),1.39(d,J=6.8Hz,3H).

[0309] Example 29: (R)-3-methyl-4-(2-(6-methyl-1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (Compound 29)

[0310]

[0311] Step 1: (R)-3-methyl-4-(7-(methylsulfonyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine

[0312] Add (R)-4-(2-chloro-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (200 mg, 0.60 mmol), biboronic acid pinacol ester (384 mg, 1.51 mmol), Pd(dppf)Cl2 (88 mg, 0.12 mmol) and potassium acetate (245 mg, 2.50 mmol) to the reaction bottle, and then add 1,4-dioxane (2 mL). Degas with nitrogen and heat the above mixture to 100 ° C under nitrogen protection, and stir for 2 hours. The reaction solution was cooled to room temperature, ethyl acetate (80 mL) was added, and the resulting mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain the crude product of the title compound (252 mg). It was not further purified and used directly in the next step.

[0313] Step 2: (R)-3-methyl-4-(2-(6-methyl-1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine

[0314] Add (R)-3-methyl-4-(7-(methylsulfonyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)morpholine (126 mg crude product, calculated as 0.30 mmol), 4-bromo-6-methyl-1H-indole (25 mg, 0.12 mmol), Pd(dppf)Cl2 (17.3 mg, 0.024 mmol) and sodium carbonate (50 mg, 0.47 mmol) to the reaction flask, and then add 1,4-dioxane (5 mL) and water (1 mL). Degas with nitrogen and heat the mixture to 85°C under nitrogen protection, and stir for 2 hours. Cool the reaction solution to room temperature, add dichloromethane (40 mL), and then wash with water. Separate the liquid and dry the organic phase with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative HPLC to obtain compound 29 (26 mg, yield 51%).

[0315] LC-MS (ESI) m / z 426.2 [M+H] + .

[0316] 1 H NMR (400MHz, DMSO-d6) δ11.08(s,1H),7.96(d,J=1.5Hz,1H),7.51(d,J=4.1Hz,1H),7 .50–7.47(m,1H),7.36(t,J=2.7Hz,1H),7.32(d,J=1.5Hz,1H),6.96(d,J=4.1Hz,1H), 4.88–4.78(m,1H),4.60–4.50(m,1H),4.05(dd,J=11.2,3.5Hz,1H),3.86–3.71(m,5H) ,3.60(td,J=11.7,2.9Hz,1H),3.52–3.41(m,1H),2.48(s,3H),1.35(d,J=6.8Hz,3H).

[0317] Example 30: (R)-4-(4-(3-methylmorpholinyl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-1H-indole-6-carbonitrile (Compound 30)

[0318]

[0319] The reaction was carried out as described above (similar to the second step of Example 29) to obtain compound 30 (30 mg, yield 46%).

[0320] LC-MS (ESI) m / z 437.1 [M+H] + .

[0321] 1 H NMR (400MHz, DMSO-d6) δ11.87(s,1H),8.36(d,J=1.5Hz,1H),8.02(t,J=1.2Hz,1H ),7.80(t,J=2.8Hz,1H),7.75–7.71(m,1H),7.57(d,J=4.1Hz,1H),7.01(d,J=4.2 Hz,1H),4.90–4.78(m,1H),4.62–4.48(m,1H),4.11–4.01(m,1H),3.86–3.70(m,5 H),3.60(td,J=11.5,11.1,2.5Hz,1H),3.54–3.44(m,1H),1.36(d,J=6.7Hz,3H).

[0322] Example 31: (R)-4-(2-(6-chloro-1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 31)

[0323]

[0324] The reaction was carried out as described above (similar to the second step of Example 29) to obtain compound 31 (18 mg, yield 27%).

[0325] LC-MS (ESI) m / z 446.1 [M+H] + .

[0326] 1 H NMR (400MHz, DMSO-d6) δ11.41(s,1H),8.08(d,J=2.0Hz,1H),7.59(t,J=2.5Hz,1H) ,7.57(d,J=1.6Hz,1H),7.55(d,J=4.1Hz,1H),7.52(t,J=2.8Hz,1H),6.99(d,J=4.1 Hz,1H),4.88–4.76(m,1H),4.58–4.46(m,1H),4.05(dd,J=11.0,2.8Hz,1H),3.86–3 .70(m,5H),3.60(td,J=11.8,2.4Hz,1H),3.54–3.43(m,1H),1.36(d,J=6.8Hz,3H).

[0327] Example 32: (R)-4-(2-(6-methoxy-1H-indol-4-yl)-7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 32)

[0328]

[0329] The reaction was carried out as described above (similar to the second step of Example 29) to obtain compound 32 (30 mg, yield 25%).

[0330] LC-MS (ESI) m / z 442.2 [M+H] + .

[0331] 1 H NMR (400MHz, DMSO-d6) δ11.04(s,1H),7.78(d,J=2.4Hz,1H),7.52(d,J=4.1Hz,1H),7.4 9–7.45(m,1H),7.31(t,J=2.7Hz,1H),7.06(dd,J=2.3,0.9Hz,1H),6.97(d,J=4.2Hz,1H ),4.89–4.77(m,1H),4.57–4.45(m,1H),4.04(dd,J=11.0,3.0Hz,1H),3.83(s,3H),3.8 1–3.71(m,5H),3.60(td,J=11.6,2.8Hz,1H),3.53–3.43(m,1H),1.35(d,J=6.7Hz,3H).

[0332] Example 33: (R)-4-(7-(ethylsulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-methylmorpholine (Compound 33)

[0333]

[0334] The reaction was carried out as shown above (similar to the second and third steps of Example 17) to obtain compound 33 (195 mg, the yields of the two steps were 92% and 79%, respectively).

[0335] LC-MS (ESI) m / z 427.2 [M+H] + .

[0336] 1H NMR (400MHz, DMSO-d6) δ11.78(s,1H),8.34(d,J=5.0Hz,1H),8.04(d,J=5.0Hz,1H ),7.63–7.57(m,2H),7.50(dd,J=3.4,2.0Hz,1H),7.03(d,J=4.1Hz,1H),4.92–4. 80(m,1H),4.62–4.50(m,1H),4.09–3.95(m,3H),3.85–3.71(m,2H),3.61(td,J=1 1.6, 2.7Hz, 1H), 3.55–3.45 (m, 1H), 1.37 (d, J = 6.8Hz, 3H), 1.12 (t, J = 7.3Hz, 3H).

[0337] Example 34: (R)-N,N-dimethyl-4-(3-methylmorpholinyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-7-sulfonamide (Compound 34)

[0338]

[0339] The reaction was carried out as shown above (similar to the second and third steps of Example 17) to obtain compound 34 (40 mg, the yields of the two steps were 13% and 32% respectively).

[0340] LC-MS (ESI) m / z 442.2 [M+H] + .

[0341] 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.33(d,J=5.0Hz,1H),7.99(d,J=5.0 Hz,1H),7.62–7.56(m,2H),7.53(dd,J=3.4,2.0Hz,1H),6.96(d,J=4.1Hz,1H ),4.90–4.80(m,1H),4.60–4.50(m,1H),4.08–4.00(m,1H),3.84–3.71(m,2H ),3.65–3.56(m,1H),3.55–3.45(m,1H),2.92(s,6H),1.37(d,J=6.7Hz,3H).

[0342] Comparative Example

[0343] Compounds 1' to 7' having the structures shown in Table 1 were synthesized as comparative example compounds.

[0344] Table 1 Comparative Example Compounds

[0345]

[0346]

[0347] Compound 1' is a known compound, which was prepared according to the synthetic route described in the literature (ACS Med. Chem. Lett. 2015, 6, 42-46).

[0348] Compound 2' was prepared according to the following synthetic route:

[0349]

[0350] Compound 3' was prepared according to the following synthetic route:

[0351]

[0352] Compound 4' was prepared according to the following synthetic route:

[0353]

[0354] Compound 5' was prepared according to the following synthetic route:

[0355]

[0356] Compound 6' was prepared according to the following synthetic route:

[0357]

[0358] Compound 7' is a known compound, prepared by referring to the synthetic route described in the literature (ACS Med. Chem. Lett. 2015, 6, 42-46). Compound 7' is a mixture of two diastereomers (ratio is about 1:1), and the following two diastereomers were obtained by HPLC chiral separation, and the nuclear magnetic resonance characteristics are as follows:

[0359]

[0360] Diastereoisomer 1: 1H NMR(400MHz,Chloroform-d)δ8.84(s,1H),8.80(br s,1H),8.72(s,1H),8.04(s,1H),7.50(d,J=3.1Hz,1H),7.23(s,1H),7.09(d,J=3.1Hz,1H ),5.62–5.55(m,1H),5.51(q,J=7.1Hz,1H),5.05(d,J=13.4Hz,1H),4.07(dd,J=11.2,3.4 Hz,1H),3.91(dd,J=11.4,3.1Hz,1H),3.84(d,J=11.3Hz,1H),3.74(td,J=11.7,2.8Hz,1H ), 3.59(td,J=12.9,3.5Hz,1H),2.72(s,3H),2.11(d,J=7.1Hz,3H),1.44(d,J=6.8Hz,3H).

[0361] Diastereoisomer II: 1 H NMR(400MHz,Chloroform-d)δ8.97(br s,1H),8.84(s,1H),8.72(s,1H),8.05(s,1H),7.50(d,J=3.2Hz,1H),7.23(s,1H),7.09(d,J= 3.1Hz,1H),5.64–5.56(m,1H),5.52(q,J=7.0Hz,1H),5.02(d,J=13.5Hz,1H),4.07(dd,J=11. 2,3.5Hz,1H),3.91(dd,J=11.4,3.1Hz,1H),3.84(d,J=11.3Hz,1H),3.75(td,J=11.8,2.8Hz, 1H),3.58(td,J=12.9,3.6Hz,1H),2.69(s,3H),2.11(d,J=7.1Hz,3H),1.44(d,J=6.8Hz,3H).

[0362] Biological test cases

[0363] In conventional biological tests and the following test examples, the following abbreviations have the following meanings:

[0364] Abbreviation meaning

[0365] ATP Adenosine triphosphate

[0366] HEPES 4-Hydroxyethylpiperazineethanesulfonic acid

[0367] Brij-35 Polyoxyethylene Laureth Ether

[0368] EDTA Ethylenediaminetetraacetic acid

[0369] DMSO Dimethyl sulfoxide

[0370] Test Example 1: In vitro ATR kinase inhibition assay

[0371] Experimental method: Caliper EZReader based on microfluidic chip technology was used to detect the mobility of the cells at ATP concentration of K m In the case of , the inhibitory activity of the compounds against ATR kinase in vitro was determined by mobility shift assay.

[0372] Experimental materials and equipment: ATR kinase (Eurofins, product catalog number 14-953); 5-FAM-AK-17 (Gill Biochemical, product catalog number 524315); 1x kinase buffer (50 mM HEPES pH 7.5, 0.0015% Brij-35, 1 M MnCl2); stop buffer (100 mM HEPES pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM EDTA); Caliper EZ Reader (Caliper Life Sciences).

[0373] Experimental steps: (1) Dissolve the test compound in DMSO and dilute it 4 times continuously for a total of 10 concentrations; (2) Use the Echo pipetting system to transfer 0.06 μL of the compound of different concentrations to a 384-well plate; (3) Use ATR kinase and 1 times kinase buffer to prepare a 2x kinase solution with an ATR kinase concentration of 10 nM; Use 5-FAM-AK-17, ATP and 1 times kinase buffer to prepare a 2x substrate solution with a 5-FAM-AK-17 concentration of 6 μM and an ATP concentration of 4 μM; (4) Add 10 μL of the 2x kinase solution to the above 384-well plate and incubate at room temperature for 10 minutes; then add 10 μL of the 2x substrate solution and incubate at 28°C for 4 hours; (5) Add 30 μL of stop buffer to terminate the reaction, then place the 384-well plate into the Caliper EZ Reader to read the conversion rate data.

[0374] Data processing: (1) Calculate the inhibition rate by conversion rate: "max" is the conversion rate of the DMSO blank control group (no compound added), "min" is the conversion rate of the enzyme-free control group (no ATR kinase added), and "conversion" is the conversion rate of the compound test group. Inhibition rate = (max-conversion) / (max-min) × 100%; (2) Use the Excel plug-in XLFit (version 5.4.0.8) to fit the curve and calculate the IC 50 value.

[0375] Experimental results: The inhibitory effects of the example compounds on ATR kinase are shown in Table 2.

[0376] Table 2

[0377]

[0378] From the above results, it can be seen that the example compounds have a good inhibitory effect on ATR kinase.

[0379] The inhibitory effects of comparative example compounds 1′ to 6′ on ATR kinase are shown in Table 3.

[0380] Table 3

[0381] Comparative Example Compound No. <![CDATA[ATR IC 50 (nM)]]> Compound 1' 56 Compound 2' 63 Compound 3' 31 Compound 4' 20 Compound 5' 15 Compound 6' 268 Compound 7' 28

[0382] Test Example 2: Cell proliferation inhibition experiment

[0383] Experimental method: The CellTiter-Glo cell viability assay kit was used to test the inhibitory effect of the compounds on the proliferation of human colon cancer cells LoVo in vitro.

[0384] Experimental materials: LoVo cells (ATCC, product catalog number CCL-29); F-12K medium (Invitrogen, product catalog number 21127-022); fetal bovine serum (Biological Industries, product catalog number 04-002-1A); GlutaMAX (Gibco, product catalog number 35050-061); 0.25% trypsin-EDTA (Invitrogen, product catalog number 25200-072); CellTiter-Glo kit (Promega, product catalog number G7558).

[0385] Experimental steps: (1) F-12K culture medium, fetal bovine serum, and GlutaMAX were prepared into complete culture medium for culturing LoVo cells; (2) LoVo cells in good growth state were taken, digested with trypsin, transferred to a centrifuge tube, centrifuged, the supernatant was discarded, fresh culture medium was added to resuspend the cells, and the cells were counted using a Vi-CELL XR cell counter. The density of the cell suspension was adjusted to 25,000-30,000 cells / mL, and the cell suspension was added to a 96-well cell culture plate at 100 μL / well. At the same time, a blank control group without cells was set up, and the culture plate was placed in a carbon dioxide incubator and incubated overnight; (3) HP was used to The D300 ultra-micropipette dispenses the DMSO solution of the test compound into the above 96-well cell culture plate, adding 0.5 μL to each well, with 10 concentrations (3-fold dilution) of each compound. A DMSO control group without compound is set up, and the culture plate is incubated in a carbon dioxide incubator for 96 hours; (4) The culture plate is taken out, equilibrated to room temperature, 100 μL of CellTiter-Glo reagent is added to each well, shaken for 10 minutes in the dark, and incubated for 10 minutes; (5) The culture plate is placed in an EnSpire plate reader and the luminescence value RLU is recorded.

[0386] Data processing: (1) Calculate the inhibition rate by luminescence value: "RLU DMSO ” is the luminescence value of the DMSO control group without adding compounds, “RLU blank ” is the luminescence value of the blank control group without cells, and “RLU compound ” is the luminescence value of the compound test group, inhibition rate = [1-(RLU compound -RLU blank ) / (RLU DMSO -RLU blank )]×100%; (2) Use Excel plug-in XLFit to fit the curve and calculate IC 50 value.

[0387] Experimental results: The inhibitory effects of the example compounds on the proliferation of LoVo cells in vitro are shown in Table 4.

[0388] Table 4

[0389]

[0390] It can be seen from the above results that the compounds of the embodiments of the present invention have a good inhibitory effect on the proliferation of LoVo cells.

[0391] The inhibitory activity of Example Compound 1 of the present invention on ATR kinase and the inhibitory activity on LoVo cell proliferation were compared with Comparative Example Compounds 4' and 5', as shown in Table 5.

[0392] Table 5

[0393] Compound No. <![CDATA[ATR IC 50 (nM)]]> <![CDATA[ATR IC 50 Ratio]]> <![CDATA[LoVo IC 50 (μM)]]> <![CDATA[LoVo IC 50 Ratio]]> Compound 4' 20 20 / 5.5＝3.64 2.34 2.34 / 0.66＝3.55 Compound 5' 15 15 / 5.5＝2.73 1.88 1.88 / 0.66＝2.85 Example Compound 1 of the present invention 5.5 / 0.66 /

[0394] The above description is only a specific implementation of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, The compound is selected from:

2. A method for preparing the compound according to claim 1 or a pharmaceutically acceptable salt thereof, the method comprising: An intermediate compound having a structure shown in formula II and R 1 -M undergoes a coupling reaction to prepare a compound having structural formula I; Among them, R 3 Halogen or R 3 When it is a halogen, M is R 3 for When M is halogen.

3. The method according to claim 2, wherein: R 3 is chlorine, M is 4. The method according to claim 2, wherein: R 3 for M is bromine.

5. A pharmaceutical composition comprising: The compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 5 in the preparation of a medicament for inhibiting ATR kinase.

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