Pyrimidino ring compounds and derivatives thereof, methods of preparation, pharmaceutical compositions, and uses

Abstract

The application discloses a pyrimidinocycle compound and a derivative thereof, a preparation method, a pharmaceutical composition and application. The compound structure is as shown in formula I, and the pyrimidinocycle compound derivative relates to a pharmaceutically acceptable salt or stereoisomer of the compound. The pyrimidinocycle compound and the derivative thereof have a high inhibitory effect on ATR, and can be used for preparing a medicine for treating diseases related to ATR mediation. The prepared medicine can exert a drug effect at a molecular level and a cell level, meanwhile, the compound has metabolic stability, is beneficial to drug preparation, and solves problems of poor curative effect, high drug dosage and great toxicity of an existing medicine. In addition, the synthesis method of the compound is widely applicable and simple to operate.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of pyrimidine ring compound and its derivative, preparation method, pharmaceutical composition and application, and more particularly to a kind of pyrimidine ring compound and its derivative, preparation method, pharmaceutical composition and application which can be prepared as efficient ATR inhibitor drug. BACKGROUND

[0002] DNA carries the genetic information necessary for synthesizing RNA and proteins, and is an essential biological macromolecule for cells to perform normal physiological functions. Various factors can induce DNA damage, such as reactive oxygen species produced by cell metabolism, replication errors, ultraviolet and ionizing radiation exposure, and certain chemicals. To cope with the threat to genome integrity, cells have a complex and precise regulatory network to respond to these damages, collectively known as the DNA damage response mechanism.

[0003] The ataxia telangiectasia mutated and Rad3-related protein kinase ATR is a key kinase in the DNA damage response mechanism, and belongs to the phosphoinositide-3-kinase-related protein family. ATR can recognize single-stranded DNA and replication stress, and then phosphorylate downstream signaling molecules to activate cell cycle checkpoints, regulate DNA replication and repair processes (homologous recombination repair, interstrand crosslink damage repair, nucleotide excision repair) to cope with DNA damage.

[0004] Cells with missing or abnormal DNA damage response factors (such as ATM, p53, etc.) are more likely to mutate into tumor cells due to genomic instability, and are more dependent on the residual ATR and its signaling pathways for DNA damage repair. The use of ATR inhibitors alone can inhibit the proliferation of these cells and induce apoptosis through "synthetic lethality" effect. In addition, ATR inhibitors can also be used in combination with various types of antitumor drugs to enhance efficacy through synergistic effect. For example, combined with platinum drugs (such as cisplatin), antimetabolites (such as fluorouracil), topoisomerase inhibitors (such as topotecan), PARP inhibitors (such as olaparib), immune checkpoint inhibitors (such as durvalumab), etc.

[0005] Currently, there is no ATR inhibitor drug approved for marketing, and four small molecule ATR inhibitors, VX-970, AZD-6738, BAY-1895344, and M4344, are only in early clinical research stage. VX-970 uses an injection as a dosage form, which is limited in clinical use. For some tumor patients, high doses of AZD6738 and BAY1895344 are required to produce efficacy, but they also produce hematological toxicity. M4344 has not yet reported clinical data. Therefore, it is still urgent to discover ATR inhibitors with novel structure, high activity and safety. SUMMARY

[0006] The present application aims to provide a pyrimidinocyclic compound and its derivative, a preparation method, a pharmaceutical composition and an application, which can be prepared into an efficient ATR inhibitor drug.

[0007] The pyrimidinocyclic compound and its derivative of the present application have the structure of formula I, and the derivative is a pharmaceutically acceptable salt or stereoisomer of the pyrimidinocyclic compound:

[0008]

[0009] wherein:

[0010] X is selected from O, S or N(R 1 );

[0011] L 1 is selected from a single bond, -CH2-, -O-, -S-, -S(O)-, -((SO)=NR 1 )-, -S(O2)-, -C(O)-, -N(R 1 )-, -N(R 1 )-CH2-, -CH=CH-, -C≡C-, -N(R 1 )-C(O)-, -C(O)-N(R 1 )-, -S(O2)N(R 1 )-, -N(R 1 )S(O2)-, -N(R 1 )C(O)N(R 1 )-, -N=S(O)R 1 - or -P(O)R 1 -;

[0012] L 2 is selected from a single bond, -O- or -N(R 1 )-;

[0013] B 1 is selected from hydrogen or C1-C6 alkyl;

[0014] B 2 is selected from C6-C 10 aryl or C5-C 10 aromatic hetero group, which C6-C 10 aryl or C5-C 10 aromatic hetero group is substituted with one or more R 2 or is unsubstituted;

[0015] B3 selected from hydrogen, halogen, cyano, hydroxyl, -NR 3 R 4 , C1-C6 alkyl, C1-C6 alkoxy, C3-C 10 heterocycloalkoxy, C3-C6 cycloalkyl, C3-C 10 heterocycloalkyl, C4-C 10 heterocycloalkenyl, C6-C 10 aryl or C5-C 10 heteroaryl; wherein said C1-C6 alkyl, C6-C 10 aryl or C5-C 10 heteroaryl is unsubstituted or substituted by one or more R 5 ;

[0016] R 1 is selected from hydrogen or C1-C6 alkyl;

[0017] R 2 is selected from hydrogen, halogen, cyano, hydroxyl, oxo, C1-C6 alkyl, -S-C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl substituted C1-C6 alkyl, amino substituted C1-C6 alkyl or NR 3 R 4 ;

[0018] R 3 , R 4 are independently from each other selected from hydrogen, C1-C6 alkylacyl or C1-C6 alkyl; or NR 3 R 4 cyclocondensed to form a 5-7 membered cyclic amine group, which is unsubstituted or substituted by C1-C6 alkyl, methylsulfonyl, ethylsulfonyl or isopropylsulfonyl, the carbon atoms of the ring of said 5-7 membered cyclic amine group being optionally substituted by O or N or being unsubstituted;

[0019] R 5 is selected from hydrogen, halogen, cyano, hydroxyl, -NR 3 R 4 , C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 alkoxy, C1-C6 alkylamino, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, -(CO)R 3 , -(CO)NR 3 R 4 , -NR 3 (CO)R 4 , -(SO2)R 3 , -(SO)R 3 , -SR 3 , -(SO2)NR 3 R4 -NR 3 (SO2)R 4 , -((SO) = NR 3 )R 4 , -P(O)R 3 -.

[0020] Preferably, in the structure of the above compounds:

[0021] X is selected from S, NH or N(CH3)-;

[0022] L 1 is selected from a single bond, -CH2-, -O-, -S-, -((SO) = NR 1 )-, -S(O2)-, -N(R 1 )-, -N(R 1 )-CH2-, -CH = CH-, -C≡C-, -N(R 1 )-C(O)-, -N(R 1 )S(O2)-, -N(R 1 )C(O)N(R 1 )-, -N = S(O)R 1 - or -P(O)R 1 -;

[0023] L 2 is selected from a single bond or -NH-;

[0024] B 1 is selected from hydrogen or methyl;

[0025] B 2 is selected from one or more R 2substituted or unsubstituted phenyl, indolinyl, pyridyl, pyrimidinyl, imidazolyl, pyrazolyl, 1H-1,2,3-triazolyl, 1H-1,2,4-triazolyl, 4H-1,2,4-triazolyl, 1H-indazolyl, 1H-indolyl, 1H-pyrrolo[2,3-b]pyridyl, 1H-pyrrolo[2,3-c]pyridyl, 1H-pyrrolo[3,2-c]pyridyl, 1H-pyrrolo[3,2-b]pyridyl, 7H-pyrrolo[2,3-c]pyridazinyl, 1H-pyrrolo[2,3-d]pyridazinyl, 1H-benzo[d]imidazolyl, 1H-benzo[d][1,2,3]triazolyl, 1H-pyrazolo[3,4-b]pyridyl, 1H-pyrazolo[3,4-c]pyridyl, 1H-pyrazolo[4,3-c]pyridyl, 1H-pyrazolo[4,3-b]pyridyl, 1H-imidazo[4,5-b]pyridyl, 3H-imidazo[4,5-b]pyridyl, 3H-imidazo[4,5-c]pyridyl, 1H-imidazo[4,5-c]pyridyl, 1H-imidazo[4,5-b]pyridyl, 7H-pyrrolo[2,3-d]pyrimidinyl, or 7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridyl;

[0026] B 3 selected from hydrogen, halogen, cyano, hydroxyl, NR 3 R 4 , C1-C6alkyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, C3-C6heterocycloalkenyl, or one or more R 5 substituted or unsubstituted phenyl, pyrrolyl, pyrazolyl, thienyl, furanyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, oxetanyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,4-dioxanyl-spiro[4.5]decanyl, or pyridinonyl;

[0027] R 1 selected from hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl;

[0028] R 2 selected from hydrogen, halogen, amino, cyano, hydroxyl, oxo, C1-C3alkyl, -S-C1-C3alkyl, C1-C3alkoxy, C1-C3haloalkyl, C1-C3haloalkoxy, C1-C3alkyl substituted with hydroxyl, aminomethyl, aminoethyl, methylamino, dimethylamino, ethylamino, or diethylamino;

[0029] R 3 , R 4 are independently of each other selected from hydrogen, C1-C3alkylacyl, or C1-C6alkyl; or NR 3R 4 ring closure to form a 5-7 membered ring amine group, which is substituted by C1-C6 alkyl, methylsulfone, ethylsulfone or isopropylsulfone or is unsubstituted, the carbon atoms of the ring of the 5-7 membered ring amine group are optionally substituted by O, N or are unsubstituted;

[0030] R 5 selected from the group consisting of hydrogen, halogen, cyano, hydroxy, -NR 3 R 4 , methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, isopropoxy, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxyethyl, ethoxyethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, pyrrolyl, phenyl, -(CO)R 3 , -(CO)NR 3 R 4 , -NR 3 (CO)R 4 , -(SO2)R 3 , -(SO)R 3 , -SR 3 , -(SO2)NR 3 R 4 , -NR 3 (SO2)R 4 , -((SO)=NR 3 )R 4 , -N=(SO)R 3 R 4 or -P(O)R 3 -.

[0031] Further preferred, in the structure of the above compounds:

[0032] X is selected from S, NH or N(CH3)-;

[0033] L 1 is selected from a single bond, -CH2-, -O-, -S-, -((SO)=NH)-, -S(O2)-, -N(R 1 )-, -NH-CH2-, -CH=CH-, -C≡C-, -N(H)-C(O)-, -N(H)S(O2)-, -N(H)C(O)N(H)-, -N=S(O)R 1 - or -P(O)R 1 -;

[0034] L 2 is selected from a single bond or -NH-;

[0035] B 1 is selected from hydrogen or methyl;

[0036] B 2 Selected from any of the following groups:

[0037]

[0038] B 3 Selected from hydrogen, halogen, cyano, hydroxyl, NR 3 R 4 , methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydro-2H-thiopyran 1,1-dioxide, piperidinyl, tetrahydropyrrolyl, 3,6-dihydro-2H-pyranyl, 1,2,3,6-tetrahydropyridinyl, 3,6-dihydro-2H-thiopyranyl or one or more R 5 any of the following groups, substituted or unsubstituted: phenyl, pyrrolyl, pyrazolyl, thienyl, furyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperidinyl, epoxybutanyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,4-dioxane-spiro[4.5]decyl or pyridonyl;

[0039] R 1 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl;

[0040] R 2 is selected from hydrogen, halogen, amino, cyano, hydroxy, oxo, -S-CH3, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, isopropoxy, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxyethyl, ethoxyethyl, hydroxymethyl, hydroxyethyl, aminomethyl, aminoethyl, methylamino, dimethylamino, ethylamino or diethylamino;

[0041] R 3 、R 4 are independently selected from hydrogen, methyl, ethyl or acetyl; or NR 3 R 4 Cyclization forms any of the following cyclic amine groups; the cyclic amine group is substituted or unsubstituted with a methyl group, an ethyl group, or a methylsulfone group;

[0042]

[0043] R 5 Selected from hydrogen, halogen, cyano, hydroxyl, -NR 3 R 4methyl, ethyl, n-propyl, i-propyl, methoxy, ethoxy, i-propoxy, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxyethyl, ethoxyethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, aminomethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, pyrrolyl, phenyl, -(CO)R 3 3 R 4 3 R 4 3 R 3 3 R 3 4 R 3 4 R 3 4 R 3 4 R 3

[0044] More preferably, in the structure of the above compounds:

[0045] X is selected from S, NH or N(CH3)-;

[0046] L 1 is selected from a single bond, -CH2-, -O-, -S-, -((SO)=NH)-, -S(O2)-, -N(R 1 )-, -NH-CH2-, -CH=CH-, -C≡C-, -N(H)-C(O)-, -N(H)S(O2)-, -N(H)C(O)N(H)-, -N=S(O)R 1 - or -P(O)R 1 -;

[0047] L 2 is selected from a single bond or -NH-;

[0048] B 1 is selected from hydrogen or methyl;

[0049] B 2 is selected from any of the following groups:

[0050]

[0051] B 3 is selected from hydrogen, halogen, cyano, hydroxy, NR 3 R 4 ​​​​​​​​​, methyl, ethyl, cyclopropyl, tetrahydropyranyl, piperidinyl, 3,6-dihydro-2H-pyranyl, 1,2,3,6-tetrahydropyridinyl or substituted or unsubstituted phenyl, pyrrolyl, pyrazolyl, thienyl, furanyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyridonyl; 5 substituted or unsubstituted phenyl, pyrrolyl, pyrazolyl, thienyl, furanyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyridonyl;

[0052] R 1 is selected from hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl;

[0053] R 2 is selected from hydrogen, halogen, amino, hydroxyl, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, hydroxymethyl, hydroxyethyl, aminomethyl, aminoethyl, methylamino, dimethylamino, ethylamino or diethylamino;

[0054] R 3 , R 4 are independently from each other selected from hydrogen, methyl, ethyl; or NR 3 R 4 are cyclized to form any of the following cyclic amine groups; the cyclic amine groups are substituted or unsubstituted by methyl, ethyl, or methylsulfonyl;

[0055]

[0056] R 5 is selected from hydrogen, halogen, cyano, hydroxyl, -NR 3 R 4 , methyl, ethyl, n-propyl, i-propyl, methoxy, ethoxy, i-propoxy, trifluoromethyl, trifluoromethoxy, hydroxymethyl, hydroxyethyl, aminomethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyranyl, piperidinyl, pyrrolyl, phenyl, -(CO)R 3 , -(CO)NR 3 R 4 , -NR 3 (CO)R 4 , -(SO2)R 3 , -(SO)R 3 , -SR 3 , -(SO2)NR 3 R 4 , -NR 3 (SO2)R 4 , -((SO) = NR 3 )R 4 , -N = (SO)R 3 R 4 or -P(O)R 3 -.

[0057] Most preferably, the above-mentioned pyrimidino ring compound is specifically any one of the following compounds:

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064] The present application includes the free form of the compounds of Formula I, as well as pharmaceutically acceptable salts and stereoisomers thereof. The pharmaceutically acceptable salts of the present application can be conveniently prepared using conventional methods. In one aspect, salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as salts derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzoic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, and the like, are included. In another aspect, salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion-exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydroxycobal, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

[0065] It should be noted that the compounds of the present application are potentially internal salts or zwitterions, since deprotonated acidic moieties, such as carboxyl groups, in the compounds can be anionic under physiological conditions, and this charge can then be counterbalanced by an internally protonated or alkylated basic moiety, such as a quaternary nitrogen atom.

[0066] As a second aspect of the present application, the method for preparing the pyrimidino ring compound and its derivatives of the present application is any one of the following methods:

[0067] (1) When X is selected from S, O or N(R 1 ), R 1 is selected from C1-C6 alkyl, the compound a containing leaving groups M 1 and M 2 is subjected to coupling reaction to obtain the compound I, respectively:

[0068]

[0069] (2) When X is N(R 1 ), R 1 is selected from hydrogen, the compound a containing leaving groups M 1 and M 2 is subjected to protection, coupling and deprotection reaction to obtain the compound I, respectively:

[0070]

[0071] wherein, L 1 , L 2 , B 1 , B 2 , B 3 are defined as described above, M 1 , M 2 are selected from halogen, methylsulfonyl or methylsulfoxide, preferably chlorine; Y is selected from p-toluenesulfonyl, tert-butyloxycarbonyl or tetrahydro-2H-pyran-2-yl, preferably p-toluenesulfonyl;

[0072] The corresponding acid or base is salted with the compound I obtained by the above method to obtain the pharmaceutically acceptable salt of the pyrimidino ring compound.

[0073] As a third aspect of the present application, the pharmaceutical composition of the present application comprises the pyrimidino ring compound and / or its derivatives and a pharmaceutically acceptable carrier.

[0074] Examples of the pharmaceutically acceptable carrier 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, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0075] The compound of the present application can form a complex with a macromolecular compound or a polymer through non-bonding interaction; or the compound of the present application as a small molecule can also be connected with a macromolecular compound or a polymer through chemical bond. The macromolecular compound can be a biological macromolecule such as a polysaccharide, a protein, a nucleic acid, a polypeptide, etc.

[0076] The mode of administration of the active ingredient or the pharmaceutical composition of the present application is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), etc.

[0077] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0078] As a fourth aspect to which the present application relates, the pyrimidino ring compound and derivatives thereof of the present application are applied to the preparation of ATR inhibitor drugs and drugs for preventing and / or treating ATR kinase-mediated diseases; and are particularly applied to cancer, including breast cancer, multiple myeloma, bone cancer, bladder cancer, cervical cancer, non-small cell lung cancer, small cell lung cancer, bronchiolar alveolar carcinoma, ovarian cancer, esophageal cancer, colorectal cancer, liver cancer, head and neck tumor, kidney cancer, liver cancer, gastrointestinal tissue cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, testicular cancer, cholangiocarcinoma, or leukemia.

[0079] The compound of the present application can be administered alone or in combination with other therapeutic agents (e.g., hypoglycemic agents). The compound of Formula I can be used in combination with other agents known to treat or ameliorate similar conditions, such as estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic / cytostatic agents, antiproliferative agents, protein kinase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, inhibitors of cell proliferation and survival signals, agents that interfere with cell cycle checkpoints, and inducers of apoptosis, cytotoxic agents, tyrosine kinase inhibitors, EGFR inhibitors, VEGFR inhibitors, serine / threonine kinase inhibitors, Bcr-Abl inhibitors, c-Kit inhibitors, Met inhibitors, Raf inhibitors, MEK inhibitors, MMP inhibitors, topoisomerase inhibitors, histone deacetylase inhibitors, proteasome inhibitors, CDK inhibitors, Bcl-2 family protein inhibitors, MDM2 family protein inhibitors, IAP family protein inhibitors, STAT family protein inhibitors, PI3K inhibitors, AKT inhibitors, integrin blockers, interferon-alpha, interleukin-12, COX-2 inhibitors, p53, p53 activators, VEGF antibodies, EGF antibodies, and the like.

[0080] Advantages: Compared with the prior art, the present application has the following remarkable advantages:

[0081] (1) This type of pyrimidocyclic compound and its derivatives and pharmaceutical compositions can effectively inhibit ATR activity, IC 50 All reached nanomolar concentrations, with the optimal concentration being below 1nM. They also had inhibitory effects on tumor cells, with IC 50 All of them are less than 5μM, even less than 1μM, and the best reaches less than 0.1μM. In addition, the compounds are metabolically stable and suitable for drug development.

[0082] (2) This type of pyrimidocyclic compound and its derivatives and pharmaceutical compositions are widely used and can be prepared as drugs for treating and / or preventing ATR-mediated diseases. The drugs can exert their efficacy at both the molecular and cellular levels, and have superior therapeutic effects, with enzyme inhibitory activity reaching nanomolar concentrations and cell inhibitory activity reaching micromolar concentrations.

[0083] (3) The compound preparation method has wide applicability and is easy to operate. DETAILED DESCRIPTION

[0084] The technical solution of the present invention will be further described below in conjunction with embodiments.

[0085] The structure of the compound of the present invention is determined by hydrogen nuclear magnetic resonance spectroscopy ( 1 The purity of the compound was determined by high performance liquid chromatography (HPLC). 1 H-NMR measurements were performed using a Bruker Advance 300 nuclear magnetic resonance spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6) and deuterated chloroform (CDCl3), with tetramethylsilane (TMS) as the internal standard. Thin-layer chromatography (TLC) was performed using thin-layer chromatography silica gel plates (Yantai Jiangyou Silica Gel Development Co., Ltd.). Homemade silica gel thin-layer plates were prepared using GF254 silica gel (Qingdao Ocean Chemical Plant). Silica gel column chromatography typically used 200-300 mesh silica gel (Qingdao Ocean Chemical Plant).

[0086] Example 1: Preparation of (R)-3-methyl-4-(7-(4-(methylsulfonyl)phenyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-1)

[0087]

[0088] Step 1: Preparation of 2,4-dichloro-7-iodothieno[3,2-d]pyrimidine (Compound 1b)

[0089] Compound 1a (10 g, 48.7 mmol) was dissolved in 50 mL trifluoroacetic acid, N-iodosuccinimide (21.8 g, 97.4 mmol) was added slowly in batches, and the reaction was carried out at 45 °C for 48 h. After the completion of the reaction was monitored by TLC, the reaction was quenched by adding saturated sodium thiosulfate solution, and filtered under suction. The filter residue was transferred to a mixed solution of dichloromethane and saturated sodium carbonate solution, stirred for 1 h, filtered under suction with diatomite, washed with saturated sodium carbonate solution twice, washed with saturated brine once, dried over anhydrous sodium sulfate, filtered under suction, and evaporated to dryness. Column chromatography gave 8 g of compound 1b as a white solid in a yield of 50%. 1 H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H).

[0090] Step 2: Preparation of (R)-4-(2-chloro-7-iodothieno[3,2-d]pyrimidin-4-yl)-3- methylmorpholine (compound 1c)

[0091] Compound 1b (1.5 g, 4.53 mmol) was dissolved in 15 mL of anhydrous ethanol, (R)-3-methylmorpholine (0.91 g, 9.06 mmol) and triethylamine (1.37 g, 13.6 mmol) were added, and the reaction was carried out at reflux for 6 h. After the completion of the reaction was monitored by TLC, the filter cake was washed with anhydrous ethanol, and dried to give 1.5 g of compound 1c as a white solid in a yield of 84%. 1 H NMR (300 MHz, DMSO-d6) δ 8.59 (s, 1H), 4.71-4.57 (m, 1H), 4.41-4.28 (m, 1H), 4.06-3.94 (m, 1H), 3.83-3.74 (m, 1H), 3.72-3.65 (m, 1H), 3.56-3.44 (m, 2H), 1.35 (d, 3H).

[0092] Step 3: Preparation of (R)-4-(2-chloro-7-(4-(methylsulfonyl)phenyl)thieno[3,2- d]pyrimidin-4-yl)-3-morpholine (compound 1d)

[0093] Compound 1c (500 mg, 1.26 mmol), 4-(methylsulfonyl)benzeneboronic acid (302 mg, 1.51 mmol), and potassium carbonate (523 mg, 3.78 mmol) were dissolved in a mixed solution of 20 mL of dioxane and 4 mL of water, and purged with nitrogen three times. Palladium (1,1'-bis(diphenylphosphino)ferrocene) dichloride (46 mg, 0.06 mmol) was added, and purged with nitrogen three times. The reaction was carried out at 60 °C for 12 h. After the completion of the reaction was monitored by TLC, the solvent was evaporated, extracted with ethyl acetate and water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered under suction, and evaporated to dryness. Column chromatography gave 300 mg of compound 1d as a white solid in a yield of 56%. 1H NMR (300 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.17 (d, J = 8.5 Hz, 2H), 8.05 (d, 2H), 4.77 - 4.66 (m, 1H), 4.45 - 4.33 (m, 1H), 4.08 - 3.97 (m, 1H), 3.85 - 3.76 (m, 1H), 3.74 - 3.66 (m, 1H), 3.61 - 3.44 (m, 2H), 3.27 (s, 3H), 1.37 (d, J = 6.8 Hz, 3H).

[0094] Step 4: Preparation of (R)-3-methyl-4-(7-(4-(methylsulfonyl)phenyl)-2-(1H- pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-1)

[0095] Compound Id (260 mg, 0.61 mmol), 7-azaindole-4-boronic acid pinacol ester (180 mg, 0.73 mmol), potassium carbonate (254 mg, 1.84 mmol) were dissolved in a mixture of 20 mL of N,N-dimethylformamide and 4 mL of water, replaced with nitrogen for three times, added 1,1'- bis(diphenylphosphino)ferrocene palladium dichloride (46 mg, 0.06 mmol), replaced with nitrogen for three times, reacted at 120 °C for 12 hours. After monitoring the completion of the reaction by TLC, the solvent was evaporated and extracted with ethyl acetate and water, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate and evaporated after suction filtration, column chromatography to obtain 120 mg of compound as a white solid, yield 39%. 1 H NMR (300 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.17 (d, J = 8.5 Hz, 2H), 8.05 (d, 2H), 4.77 - 4.66 (m, 1H), 4.45 - 4.33 (m, 1H), 4.08 - 3.97 (m, 1H), 3.85 - 3.76 (m, 1H), 3.74 - 3.66 (m, 1H), 3.61 - 3.44 (m, 2H), 3.27 (s, 3H), 1.37 (d, J = 6.8 Hz, 3H).

[0096] Using the synthesis method of compound I-1, intermediates 1c and the corresponding boronic acid or boronic acid ester were used to obtain compounds I-2 to I-36, I-68 to I-72, I-75 to I-140 through two-step coupling reaction.

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109] Example 2: Preparation of (R,E)-3-methyl-4-(7-(2-(pyridin-2-yl)vinyl)-2-(lH- pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-37)

[0110]

[0111] Step 1: Preparation of (R,E)-4-(2-chloro-7-(2-(pyridin-2-yl)vinyl)thieno[3,2- d]pyrimidin-4-yl)-3-methylmorpholine (Compound le)

[0112] Compound lc (500 mg, 1.26 mmol) was dissolved in 20 mL DMF, 2-vinylpyridine (265 mg, 2.52 mmol), and DIPEA (489 mg, 3.78 mmol) were added, after nitrogen replacement for three times, palladium acetate (14 mg, 0.06 mmol) and 3-(o-methylphenyl)phosphine (38 mg, 0.12 mmol) were added, after nitrogen replacement for three times again, 100 °C reaction overnight. After monitoring the reaction completion by TLC, appropriate amount of water was added and extracted with ethyl acetate, the organic layer was washed with water for four times and saturated brine once, dried over anhydrous sodium sulfate, filtered and evaporated, column chromatography gave 300 mg of compound le as a yellow solid, yield 64%. 1H NMR (300 MHz, DMSO-d6) δ 8.63 - 8.58 (m, 2H), 7.91 (d, J = 16.2 Hz, 1H), 7.84 - 7.76 (m, 1H), 7.69 (d, J = 16.2 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 7.32 - 7.25 (m, 1H), 4.74 - 4.62 (m, 1H), 4.42 - 4.32 (m, 1H), 4.06 - 3.95 (m, 1H), 3.83 - 3.66 (m, 2H), 3.61 - 3.42 (m, 2H), 1.36 (d, J = 6.8 Hz, 3H).

[0113] Step 2: Preparation of (R,E)-3-methyl-4-(7-(2-(pyridin-2-yl)vinyl)-2-(lH- pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (1-37)

[0114] Preparation Method same as Step 4 in Example 1, the product was a light yellow solid, yield 45%. 1 H NMR (300 MHz, DMSO-d6) δ 8.63 - 8.58 (m, 2H), 7.91 (d, J = 16.2 Hz, 1H), 7.84 - 7.76 (m, 1H), 7.69 (d, J = 16.2 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 7.32 - 7.25 (m, 1H), 4.74 - 4.62 (m, 1H), 4.42 - 4.32 (m, 1H), 4.06 - 3.95 (m, 1H), 3.83 - 3.66 (m, 2H), 3.61 - 3.42 (m, 2H), 1.36 (d, J = 6.8 Hz, 3H).

[0115] Example 3: Preparation of (R)-4-(7-(cyclopropynyl)-2-(lH-pyrrolo[2,3-b]pyridin-4- yl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (1-38)

[0116]

[0117] Step 1: Preparation of (R)-4-(2-chloro-7-(cyclopropynyl)thieno[3,2-d]pyrimidin-4- yl)-3-methylmorpholine (Compound If)

[0118] Compound 1c (500 mg, 1.26 mmol), bis(triphenylphosphine)palladium dichloride (44 mg, 0.06 mmol), cuprous iodide (24 mg, 0.13 mmol) were added into a two-necked flask, which was replaced by nitrogen for three times. Then cyclopropylacetylene (167 mg, 2.52 mmol), DIPEA (488 mg, 3.78 mmol) and 15 mL DMF were added in sequence by syringe, and the reaction was carried out at 40 °C overnight. After TLC monitoring the reaction was completed, water and ethyl acetate were added for extraction, the organic layer was washed with water four times and saturated brine once, dried over anhydrous sodium sulfate, filtered and evaporated to dryness, and column chromatography to obtain 270 mg of compound 1f as a white solid, with a yield of 64%. 1 H NMR (300 MHz, DMSO-d6) δ 8.38 (s, 1H), 4.70-4.58 (m, 1H), 4.39-4.26 (m, 1H), 4.05-3.95 (m, 1H), 3.82-3.73 (m, 1H), 3.72-3.64 (m, 1H), 3.58-3.44 (m, 2H), 1.67-1.57 (m, 1H), 1.33 (d, J = 6.8 Hz, 3H), 0.97-0.88 (m, 2H), 0.80-0.73 (m, 2H).

[0119] Step 2: Preparation of compound (R)-4-(7-(cyclopropylacetylenyl)-2-(1H- pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-38)

[0120] Preparation method same as step 4 in Example 1. The product was a white solid with a yield of 36%. 1 H NMR (300 MHz, DMSO-d6) δ 8.38 (s, 1H), 4.70-4.58 (m, 1H), 4.39-4.26 (m, 1H), 4.05-3.95 (m, 1H), 3.82-3.73 (m, 1H), 3.72-3.64 (m, 1H), 3.58-3.44 (m, 2H), 1.67-1.57 (m, 1H), 1.33 (d, J = 6.8 Hz, 3H), 0.97-0.88 (m, 2H), 0.80-0.73 (m, 2H).

[0121] Example 4: Preparation of (R)-2-(1H-benzo[d]imidazol-1-yl)-4-(3- methylmorpholino)-N-(pyridin-4-yl)thieno[3,2-d]pyrimidin-7-amine (I-39)

[0122]

[0123] Step 1: Preparation of compound (R)-4-(2-(lH-benzo[d]imidazol-l-yl)-7- iodothieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (compound Ig)

[0124] Compound lc (400 mg, 1.01 mmol), benzo[d]imidazole (143 mg, 1.21 mmol) were dissolved in 15 mL DMF, cesium carbonate (986 mg, 3.03 mmol) was added, and the reaction was carried out at 110 °C for 12 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated, water and ethyl acetate were added for extraction, the organic layer was washed with water once and saturated brine once, dried over anhydrous sodium sulfate, and then evaporated after suction filtration. Column chromatography gave 320 mg of product as a white solid, with a yield of 66%. 1 H NMR (300 MHz, Chloroform-d) δ 9.07 (s, 1H), 8.93 (d, J = 8.1 Hz, 1H), 7.99 (s, 1H), 7.85 (d, J = 8.0 Hz, 1H), 7.51 - 7.33 (m, 2H), 4.86 - 4.73 (m, 1H), 4.57 - 4.43 (m, 1H), 4.22 - 4.09 (m, 1H), 3.96 - 3.81 (m, 2H), 3.80 - 3.58 (m, 2H), 1.51 (d, J = 6.8 Hz, 3H).

[0125] Step 2: Preparation of (R)-2-(lH-benzo[d]imidazol-l-yl)-4-(3-methylmorpholino)-N- (pyridin-4-yl)thieno[3,2-d]pyrimidin-7-amine (I-39)

[0126] Compound Ig (200 mg, 0.42 mmol), 4-aminopyridine (59 mg, 0.63 mmol), Pd2(dba)3 (19 mg, 0.021 mmol), Xantphos (24 mg, 0.04 mmol), cesium carbonate (411 mg, 1.26 mmol) were dissolved in 20 mL of anhydrous dioxane, replaced with nitrogen three times, and the reaction was carried out at reflux for 12 hours. After the reaction was completed by TLC monitoring, an appropriate amount of water and ethyl acetate were added for extraction, the organic layer was washed with water three times and then saturated brine once, dried over anhydrous sodium sulfate, and then evaporated after suction filtration. Column chromatography gave 95 mg of product as a white solid, with a yield of 51%. 1H NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 8.77 (s, 1H), 8.40 - 8.36 (m, 1H), 8.28 (d, J = 6.5 Hz, 2H), 7.96 (s, 1H), 7.76 - 7.73 (m, 1H), 7.33 - 7.28 (m, 2H), 7.13 (d, J = 6.5 Hz, 2H), 4.90 - 4.78 (m, 1H), 4.59 - 4.48 (m, 1H), 4.12 - 4.05 (m, 1H), 3.89 - 3.84 (m, 1H), 3.82 - 3.75 (m, 1H), 3.68 - 3.59 (m, 2H), 1.44 (d, J = 6.8 Hz, 3H).

[0127] The preparation method of compounds I-40-I-48 is similar to that of compound I-39.

[0128]

[0129]

[0130] Example 5: Preparation of (R)-4-(2-(1H-benzo[d]imidazol-1-yl)-7-(pyridin-4-yl)thieno[3,2- d]pyrimidin-4-yl)-3-methylmorpholine (I-49)

[0131] Compound 1g (200 mg, 0.42 mmol), 4-pyridine boronic acid (61 mg, 0.50 mmol), potassium carbonate (174 mg, 1.26 mmol) were dissolved in a mixed solution of 25 mL of dioxane and 5 mL of water, replaced with nitrogen for three times, and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (15 mg, 0.02 mmol) was added, and then replaced with nitrogen for three times, and reacted at 80 °C for 12 hours. After the completion of the reaction was monitored by TLC, the solvent was evaporated and extracted with ethyl acetate and water, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then evaporated by suction filtration. Column chromatography gave 87 mg of the target product as a light yellow solid in a yield of 48%. 1 H NMR (400 MHz, DMSO-d6) δ 9.18 (s, 1H), 8.80 (s, 1H), 8.75 (d, J = 6.1 Hz, 2H), 8.50 (d, J = 9.5 Hz, 1H), 8.09 - 8.04 (m, 2H), 7.77 (d, J = 7.6 Hz, 1H), 7.42 - 7.29 (m, 2H), 4.90 - 4.78 (m, 1H), 4.63 - 4.48 (m, 1H), 4.15 - 4.03 (m, 1H), 3.92 - 3.74 (m, 2H), 3.68 - 3.53 (m, 2H), 1.43 (d, J = 6.8 Hz, 3H).

[0132] Example 6: Preparation of (R)-1-(4-(3-methylmorpholino)-7-(pyridin-4-yl)thieno[3,2- d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (I-50)

[0133]

[0134] Compound I-50 was obtained by a similar procedure to Example 5 as a white solid in 30% yield. 1 H NMR (300 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.75 - 8.69 (m, 2H), 8.15 (d, J = 8.5 Hz, 1H), 7.97 - 7.90 (m, 2H), 7.61 (s, 2H), 7.24 - 7.17 (m, 1H), 7.13 - 7.05 (m, 1H), 7.01 - 6.93 (m, 1H), 4.86 - 4.73 (m, 1H), 4.47 - 4.37 (m, 1H), 4.17 - 4.04 (m, 1H), 3.92 - 3.76 (m, 2H), 3.73 - 3.57 (m, 2H), 1.45 (d, J = 6.8 Hz, 3H).

[0135] The preparation method of compounds I-73-I-74 is similar to compound I-50.

[0136]

[0137] Example 7: Preparation of (R)-4-(2-(1H-benzo[d]imidazol-1-yl)-7-(methylsulfonyl)thieno[3,2- d]pyrimidin-4-yl)-3-methylmorpholine (I-51)

[0138] Compound 1g (200 mg, 0.42 mmol), sodium methanesulfinate (342 mg, 3.35 mmol), L-valine (10 mg, 0.084 mmol), cuprous iodide (8 mg, 0.042 mmol) were dissolved in 10 mL of dimethyl sulfoxide, replaced with nitrogen for three times, and reacted at 110 °C for 12 hours. After the reaction was completed by TLC monitoring, ethyl acetate and water were added for extraction, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then filtered under suction and evaporated to dryness. Column chromatography gave 86 mg of a white solid in a yield of 48%. 1H NMR (300 MHz, DMSO-d6) δ 9.22 (s, 1H), 9.10 (s, 1H), 8.69 (d, J = 6.3 Hz, 1H), 7.78 (d, J = 7.3 Hz, 1H), 7.46 - 7.31 (m, 2H), 4.94 - 4.75 (m, 1H), 4.69 - 4.51 (m, 1H), 4.13 - 4.00 (m, 1H), 3.90 - 3.72 (m, 2H), 3.68 - 3.53 (m, 2H), 3.47 (s, 3H), 1.42 (d, J = 6.8 Hz, 3H).

[0139] Example 8: Preparation of (R)-(2-(lH-benzo[d]imidazol-l-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)dimethyl phosphine oxide (I-52)

[0140] Compound 1g (200 mg, 0.42 mmol), dimethyl phosphine oxide (50 mg, 0.63 mmol), cesium carbonate (410 mg, 1.25 mmol), Pd2(dba)3(20 mg, 0.02 mmol), Xantphos (25 mg, 0.04 mmol) were added into two flasks, nitrogen was replaced for three times, 20 mL anhydrous dioxane was added, nitrogen was replaced for three times again, the reaction was refluxed for 12 hours. TLC was used to monitor the reaction completion, the solvent was evaporated, water and ethyl acetate were used to extract, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and evaporated, column chromatography was used to get 60 mg product as white solid, yield 33%. 1 H NMR (300 MHz, DMSO-d6) δ 9.22 (s, 1H), 9.10 (s, 1H), 8.69 (d, J = 6.3 Hz, 1H), 7.78 (d, J = 7.3 Hz, 1H), 7.46 - 7.31 (m, 2H), 4.94 - 4.75 (m, 1H), 4.69 - 4.51 (m, 1H), 4.13 - 4.00 (m, 1H), 3.90 - 3.72 (m, 2H), 3.68 - 3.53 (m, 2H), 3.47 (s, 3H), 1.42 (d, J = 6.8 Hz, 3H).

[0141] Example 9: Preparation of (R)-4-(2-(lH-benzo[d]imidazol-l-yl)-7- (morpholinomethyl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (I-53)

[0142] Compound 1g (200 mg, 0.42 mmol), morpholino potassium trifluoroborate (173 mg, 0.84 mmol), cesium carbonate (410 mg, 1.25 mmol), palladium acetate (10 mg, 0.04 mmol), X-phos (30 mg, 0.63 mmol) were added into a flask in turn, which was replaced by nitrogen for three times, then 20 mL of anhydrous dioxane and 2 mL of water were added, which was replaced by nitrogen for three times again, and the reaction was refluxed for 12 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated, water and ethyl acetate were added for extraction, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then evaporated and purified by column chromatography to obtain 75 mg of the product as a white solid, with a yield of 40%. 1 H NMR (300 MHz, DMSO-d6) δ 9.20 (s, 1H), 8.71 (d, J = 7.9 Hz, 1H), 8.16 (s, 1H), 7.78 (d, J = 7.5 Hz, 1H), 7.49 - 7.29 (m, 2H), 4.92 - 4.77 (m, 1H), 4.64 - 4.47 (m, 1H), 4.10 - 4.01 (m, 1H), 3.88 - 3.71 (m, 4H), 3.68 - 3.49 (m, 6H), 2.53 - 2.51 (m, 2H), 1.40 (d, J = 6.7 Hz, 3H).

[0143] Example 10: Preparation of (R)-4-(2-(lH-benzo[d]imidazol-l-yl)-7-(pyridin-4- ylsulfanyl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (I-54)

[0144] Compound 1g (200 mg, 0.42 mmol), 4-mercaptopyridine (70 mg, 0.63 mmol), Pd2(dba)3 (19 mg, 0.021 mmol), Xantphos (24 mg, 0.04 mmol), cesium carbonate (411 mg, 1.26 mmol) were dissolved in 20 mL of anhydrous dioxane, which was replaced by nitrogen for three times, and the reaction was refluxed for 12 hours. After the reaction was completed by TLC monitoring, an appropriate amount of water and ethyl acetate were added for extraction, the organic layer was washed with water for three times, and then with saturated brine once, dried over anhydrous sodium sulfate, and then evaporated and purified by column chromatography to obtain 120 mg of the product as a white solid, with a yield of 62%. 1H NMR (300 MHz, DMSO-d6) δ 9.10 (s, 1H), 8.86 (s, 1H), 8.38 - 8.34 (m, 2H), 7.95 (d, J = 8.1 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.28 - 7.21 (m, 1H), 7.18 - 7.15 (m, 2H), 7.13 - 7.06 (m, 1H), 4.86 (s, 1H), 4.71 - 4.53 (m, 1H), 4.11 - 4.02 (m, 1H), 3.89 - 3.74 (m, 2H), 3.68 - 3.53 (m, 2H), 1.44 (d, J = 6.8 Hz, 3H).

[0145] Example 11: Preparation of (R)-4-(2-(lH-benzo[d]imidazol-l-yl)-7-(pyridin-4- ylsulfonyl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (I-55)

[0146] Compound I-54 (110 mg, 0.24 mmol) was dissolved in a mixture of 10 mL of methanol and 5 mL of water, and potassium peroxymonosulfate (440 mg, 0.72 mmol) was added. After stirring overnight at room temperature, the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated, and the product was extracted with dichloromethane and water. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and then filtered and evaporated. Column chromatography gave 54 mg of the product as a white solid in a yield of 46%. 1 H NMR (300 MHz, DMSO-d6) δ 9.10 (s, 1H), 8.86 (s, 1H), 8.38 - 8.34 (m, 2H), 7.95 (d, J = 8.1 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.28 - 7.21 (m, 1H), 7.18 - 7.15 (m, 2H), 7.13 - 7.06 (m, 1H), 4.86 (s, 1H), 4.71 - 4.53 (m, 1H), 4.11 - 4.02 (m, 1H), 3.89 - 3.74 (m, 2H), 3.68 - 3.53 (m, 2H), 1.44 (d, J = 6.8 Hz, 3H).

[0147] Example 12: Preparation of (2-(lH-benzo[d]imidazol-l-yl)-4-((R)-3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)(imino)(pyridin-4-yl)-lambda 6 sulfone (I-56)

[0148] Compound I-54 (180 mg, 0.39 mmol) was dissolved in 10 mL of methanol, and ammonium carbonate (75 mg, 0.78 mmol) and iodo-benzoic acid (251 mg, 0.78 mmol) were added successively under ice bath, and the mixture was stirred slowly to room temperature for 12 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated, and the product was extracted with dichloromethane and water. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated. The product was obtained by column chromatography in the form of a white solid at a yield of 48 mg, 25%. 1 H NMR (300 MHz, DMSO-d6) δ 9.23 (s, 1H), 9.10 (s, 1H), 8.77-8.68 (m, 3H), 8.05-7.99 (m, 2H), 7.75 (d, J = 7.5 Hz, 1H), 7.45 (d, J = 16.7 Hz, 1H), 7.35 (d, J = 16.5 Hz, 1H), 5.82 (d, J = 3.2 Hz, 1H), 4.91-4.73 (m, 1H), 4.64-4.44 (m, 1H), 4.09-3.97 (m, 1H), 3.86-3.70 (m, 2H), 3.65-3.45 (m, 2H), 1.39 (d, J = 8.4 Hz, 3H).

[0149] Example 13: Preparation of (R)-4-(2-(1H-benzo[d]imidazol-1-yl)-7-(4-methylthiazol-5-yl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (I-57)

[0150] Compound 1g (200 mg, 0.42 mmol) was dissolved in 10 mL of DMF, and 4-methylthiazole (50 mg, 0.50 mmol), potassium acetate (82 mg, 0.84 mmol), and palladium acetate (10 mg, 0.04 mmol) were added successively, and the mixture was reacted at 100°C for 12 hours. After the reaction was completed by TLC monitoring, the product was extracted with water and ethyl acetate, and the organic layer was washed with water three times and saturated brine once. After drying over anhydrous sodium sulfate, the product was evaporated, and column chromatography was performed to obtain 90 mg of the product in the form of a white solid at a yield of 48%. 1 H NMR (300 MHz, DMSO-d6) δ 9.18 (d, J = 3.8 Hz, 2H), 8.59-8.51 (m, 2H), 7.80-7.74 (m, 1H), 7.40-7.29 (m, 2H), 4.94-4.80 (m, 1H), 4.64-4.50 (m, 1H), 4.16-4.02 (m, 1H), 3.90-3.75 (m, 2H), 3.72-3.55 (m, 2H), 2.59 (s, 3H), 1.44 (d, J = 6.8 Hz, 3H).

[0151] Example 14: Preparation of (R)-N-(2-(lH-benzo[d]imidazol-l-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)isonicotinamide (I-58)

[0152]

[0153] Step 1: Preparation of compound (R)-2-(lH-benzo[d]imidazol-l-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-amine (Compound 1i)

[0154] Compound 1g (200 mg, 0.42 mmol), benzophenone imine (70 mg, 0.63 mmol), Pd2(dba)3 (19 mg, 0.021 mmol), Xantphos (24 mg, 0.04 mmol), cesium carbonate (411 mg, 1.26 mmol) were dissolved in 20 mL of anhydrous dioxane, replaced with nitrogen for three times, and refluxed for 12 hours. After the reaction was completed by TLC monitoring, the reaction mixture was extracted with a proper amount of water and ethyl acetate, the organic layer was washed with water once and saturated brine once, dried over anhydrous sodium sulfate, and then concentrated by suction filtration and dried. The residue was dissolved in 10 mL of methanol, and sodium acetate (75 mg, 0.9 mmol) and hydroxylamine hydrochloride (47 mg, 0.68 mmol) were sequentially added. After stirring at room temperature for 6 hours, the mixture was suction filtered, and the filter residue was washed with methanol and dried to obtain 90 mg of the product as a yellow solid, in a two-step yield of 59%. 1 H NMR (300 MHz, DMSO-d6) δ 9.29 (s, 1H), 8.69 (d, J = 8.3 Hz, 1H), 7.76 (d, J = 7.8 Hz, 1H), 7.45 - 7.29 (m, 2H), 6.68 (s, 1H), 5.41 (s, 2H), 4.88 - 4.74 (m, 1H), 4.57 - 4.43 (m, 1H), 4.12 - 4.00 (m, 1H), 3.88 - 3.73 (m, 2H), 3.66 - 3.49 (m, 2H), 1.40 (d, J = 6.7 Hz, 3H).

[0155] Step 2: Preparation of (R)-N-(2-(lH-benzo[d]imidazol-l-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)isonicotinamide (I-58)

[0156] Compound 1i (150 mg, 0.41 mmol) was dissolved in 10 mL of anhydrous dichloromethane, and triethylamine (125 mg, 1.23 mmol) and isonicotinyl chloride hydrochloride (110 mg, 0.61 mmol) were added successively under ice bath, and the reaction was slowly warmed to room temperature for 12 hours. After the reaction was completed by TLC monitoring, an appropriate amount of dichloromethane and water were added for extraction, the organic layer was washed with saturated brine once, dried over anhydrous sodium sulfate, and then filtered and evaporated to dryness, and column chromatography was performed to obtain 92 mg of a light yellow solid, with a yield of 48%. 1 H NMR (300 MHz, DMSO-d6) δ 10.53 (s, 1H), 9.49 (s, 1H), 8.91-8.85 (m, 2H), 8.64-8.55 (m, 2H), 7.96 (d, J = 6.1 Hz, 2H), 7.78 (d, J = 8.8 Hz, 1H), 7.44-7.31 (m, 2H), 4.93-4.75 (m, 1H), 4.60-4.46 (m, 1H), 4.16-4.07 (m, 1H), 3.93-3.76 (m, 2H), 3.71-3.59 (m, 2H), 1.45 (d, J = 6.8 Hz, 3H).

[0157] Example 15: Preparation of (R)-N-(2-(lH-benzo[d]imidazol-l-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)-2-(pyridin-4-yl)acetamide (I-59)

[0158] 4-pyridine acetate hydrochloride (150 mg, 0.864 mmol) was dissolved in 10 mL of DMF, and DIPEA (330 mg, 2.54 mmol) and HATU (547 mg, 1.44 mmol) were added successively, and stirred at room temperature for 0.5 hours, and then compound 1g (211 mg, 0.576 mmol) was added, and the reaction was continued at room temperature for 12 hours. After the reaction was completed by TLC monitoring, an appropriate amount of water and ethyl acetate were added for extraction, the organic layer was washed with water three times, and then washed with saturated brine once, dried over anhydrous sodium sulfate, and then filtered and evaporated to dryness, and column chromatography was performed to obtain 89 mg of a white solid, with a yield of 32%. 1 H NMR (300 MHz, DMSO-d6) δ 10.18 (s, 1H), 9.56 (s, 1H), 8.59-8.51 (m, 3H), 8.40 (s, 1H), 7.79 (d, J = 7.7 Hz, 1H), 7.48-7.32 (m, 4H), 4.91-4.72 (m, 1H), 4.55-4.36 (m, 1H), 4.14-4.05 (m, 1H), 4.02 (s, 2H), 3.91-3.73 (m, 2H), 3.69-3.54 (m, 2H), 1.42 (d, J = 6.8 Hz, 3H).

[0159] Example 16: Preparation of (R)-1-(2-(1H-benzo[d]imidazol-1-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)-3-(pyridin-2-yl)urea (I-60)

[0160] Compound 1i (200 mg, 0.55 mmol), 2-isocyanate pyridine (79 mg, 0.66 mmol) were dissolved in 20 mL dry dioxane and refluxed for 6 h. After completion of the reaction as monitored by TLC, the solvent was evaporated, extracted with water and ethyl acetate. The organic layer was washed once with saturated brine, dried over anhydrous sodium sulfate and evaporated. The residue was column chromatographed to get 86 mg of the product as a white solid in 32% yield. 1 H NMR (300 MHz, DMSO-d6) δ 11.62 (s, 1H), 10.17 (s, 1H), 9.27 (s, 1H), 8.72 - 8.63 (m, 1H), 8.34 - 8.23 (m, 2H), 7.86 - 7.75 (m, 2H), 7.45 - 7.28 (m, 3H), 7.13 - 7.04 (m, 1H), 4.92 - 4.74 (m, 1H), 4.63 - 4.44 (m, 1H), 4.14 - 3.99 (m, 1H), 3.90 - 3.72 (m, 2H), 3.71 - 3.51 (m, 2H), 1.42 (d, J = 6.7 Hz, 3H).

[0161] Example 17: Preparation of (R)-N-(2-(1H-benzo[d]imidazol-1-yl)-4-(3- methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)pyridine-3-sulfonamide (I-61)

[0162] Compound 1i (150 mg, 0.41 mmol), 3-pyridine sulfonyl chloride (110 mg, 0.61 mmol) were dissolved in 10 mL pyridine and stirred at room temperature for 2 h. After completion of the reaction as monitored by TLC, ethyl acetate was added and the organic layer was washed twice with 1 N hydrochloric acid and once with saturated brine. It was dried over anhydrous sodium sulfate and evaporated. The residue was column chromatographed to get 55 mg of the product as a white solid in 26% yield. 1H NMR (300 MHz, DMSO-d6) δ 10.77 (s, 1H), 9.30 (s, 1H), 9.01 - 8.96 (m, 1H), 8.67 - 8.61 (m, 1H), 8.52 - 8.46 (m, 1H), 8.27 - 8.20 (m, 1H), 8.04 - 7.97 (m, 1H), 7.75 (d, J = 7.7 Hz, 1H), 7.48 - 7.31 (m, 3H), 4.83 - 4.69 (m, 1H), 4.51 - 4.38 (m, 1H), 4.10 - 4.02 (m, 1H), 3.87 - 3.71 (m, 2H), 3.62 - 3.54 (m, 2H), 1.39 (d, J = 6.8 Hz, 3H).

[0163] Example 18: Preparation of (R)-N-methyl-l-(4-(3-methylmorpholino)-7-(pyridin-4- yl)thieno[3,2-d]pyrimidin-2-yl)-lH-benzo[d]imidazol-2-amine (I-62)

[0164] (R)-4-(2-chloro-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (200 mg, 0.58 mmol), N-methyl-lH-benzo[d]imidazol-2-amine (102 mg, 0.69 mmol), Pd2(dba)3 (26 mg, 0.03 mmol), Xantphos (28 mg, 0.06 mmol), cesium carbonate (564 mg, 1.73 mmol) were dissolved in 20 mL of anhydrous dioxane, replaced with nitrogen for three times, and refluxed for 12 hours. After the reaction was completed as monitored by TLC, the solvent was evaporated, and the product was extracted with a proper amount of water and ethyl acetate. The organic layer was washed with water once and saturated brine once, dried over anhydrous sodium sulfate, and evaporated. Column chromatography gave 92 mg of the product as a white solid in a yield of 35%. 1 H NMR (300 MHz, DMSO-d6) δ 8.80 - 8.72 (m, 3H), 8.55 - 8.47 (m, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.90 (d, J = 6.1 Hz, 2H), 7.27 (d, J = 7.6 Hz, 1H), 7.14 - 7.06 (m, 1H), 7.03 - 6.95 (m, 1H), 4.83 - 4.73 (m, 1H), 4.45 - 4.34 (m, 1H), 4.16 - 4.07 (m, 1H), 3.93 - 3.76 (m, 2H), 3.74 - 3.58 (m, 2H), 3.03 (d, J = 4.8 Hz, 3H), 1.45 (d, J = 6.8 Hz, 3H).

[0165] The preparation method of compounds I-63-I-65 is similar to that of compound I-62.

[0166]

[0167]

[0168] Example 19: Preparation of (R)-3-methyl-4-(7-(pyridin-4-yl)-2-(1H-pyrrolo[2,3- b]pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)morpholine (I-66)

[0169]

[0170] Step 1: Preparation of compound 2,4-dichloro-7-iodo-5H-pyrrolo[3,2-d]pyrimidine (1k)

[0171] Compound 1j (2 g, 10.6 mmol) was dissolved in 20 mL THF, N-iodosuccinimide (4.8 g, 21.2 mmol) was added in batches, and stirred at room temperature for 12 hours. After monitoring the completion of the reaction by TLC, the reaction was quenched by adding saturated sodium thiosulfate solution, and the solid was precipitated. After stirring for 15 minutes, the filter residue was washed with water three times and dried to obtain 2.2 g of compound 1k as a white solid with a yield of 66%. 1 H NMR (300 MHz, DMSO-d6) δ 13.22 (s, 1H), 8.31 (s, 1H).

[0172] Step 2: Preparation of compound 2,4-dichloro-7-iodo-5-p-tolylsulfonyl-5H-pyrrolo[3,2- d]pyrimidine (1l)

[0173] Compound 1k (2 g, 6.37 mmol) was dissolved in 20 mL DMF, and cesium carbonate (6.2 g, 19.1 mmol) and p-toluenesulfonyl chloride (2.4 g, 12.7 mmol) were added in sequence, and stirred at room temperature for 12 hours. After monitoring the completion of the reaction by TLC, an appropriate amount of water was added, and extracted with ethyl acetate. The organic layer was washed with water three times, and then with saturated brine. After drying over anhydrous sodium sulfate, the filtrate was evaporated, and column chromatography was performed to obtain 2 g of compound 1l as a white solid with a yield of 69%. 1 H NMR (300 MHz, Chloroform-d) δ 8.43 (s, 1H), 7.77 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 8.0 Hz, 3H), 2.44 (s, 3H).

[0174] Step 3: Preparation of compound (R)-4-(2-chloro-7-iodo-5-p-tolylsulfonyl- 5H-pyrrolo[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (1m)

[0175] Compound 1m was prepared from compound 1l by the method of step 2 in Example 1 as a white solid in 57% yield. 1 H NMR (300 MHz, DMSO-d6) δ 8.26 (s, 1H), 7.51 - 7.45 (m, 2H), 7.34 - 7.28 (m, 2H), 4.68 - 4.57 (m, 1H), 3.97 - 3.67 (m, 4H), 3.63 - 3.42 (m, 2H), 2.32 (s, 3H), 1.20 (d, J = 7.5 Hz, 3H).

[0176] Step 4: Preparation of compound (R)-4-(2-chloro-7-(pyridin-4-yl-5-p-tolylsulfonyl- 5H-pyrrolo[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (1n)

[0177] Compound 1n was prepared from compound 1m by the method of step 3 in Example 1 as a white solid in 63% yield. 1 H NMR (300 MHz, Chloroform-d) δ 8.65 (d, J = 5.3 Hz, 2H), 7.97 (s, 1H), 7.81 (d, J = 4.9 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 7.13 (d, J = 8.0 Hz, 2H), 4.80 - 4.67 (m, 1H), 4.08 - 3.97 (m, 2H), 3.97 - 3.86 (m, 1H), 3.84 - 3.60 (m, 3H), 2.32 (s, 3H), 1.29 (d, J = 6.8 Hz, 3H).

[0178] Step 5: Preparation of (R)-3-methyl-4-(7-(pyridin-4-yl)-2-(lH-pyrrolo[2,3-b]pyridin-4- yl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)morpholine (I-66)

[0179] Compound 1n was prepared from compound 1m by the method of step 4 in Example 1 as a yellow solid in 36% yield. 1H NMR (300 MHz, DMSO-d6) δ 12.00 (s, 1H), 11.72 (s, 1H), 8.60 (m, J = 6.0 Hz, 2H), 8.44 - 8.32 (m, 4H), 8.09 (m, J = 5.0 Hz, 1H), 7.64 - 7.56 (m, 1H), 7.44 - 7.38 (m, 1H), 4.82 - 4.72 (m, 1H), 4.30 - 4.21 (m, 1H), 4.10 - 4.00 (m, 1H), 3.88 - 3.79 (m, 2H), 3.74 - 3.53 (m, 2H), 1.33 (d, J = 6.7 Hz, 3H).

[0180] Example 20: Preparation of (R)-3-methyl-4-(5-methyl-7-(pyridin-4-yl)-2-(1H-pyrrolo[2,3- b]pyridin-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)morpholine (I-67)

[0181] Compound I-67 was obtained by a similar method of preparation of compound I-66 as a yellow solid in 56% yield. 1 H NMR (300 MHz, DMSO-d6) δ 11.78 (s, 1H), 8.67 - 8.59 (m, 2H), 8.53 (s, 1H), 8.42 - 8.36 (m, 1H), 8.32 - 8.24 (m, 2H), 8.18 - 8.12 (m, 1H), 7.67 - 7.60 (m, 1H), 7.48 - 7.40 (m, 1H), 4.13 (s, 3H), 4.00 - 3.88 (m, 3H), 3.87 - 3.78 (m, 1H), 3.61 - 3.50 (m, 2H), 3.25 - 3.12 (m, 1H), 1.11 (d, J = 6.1 Hz, 3H).

[0182] Example 21: Inhibition effect of compounds on ATR kinase

[0183] (1) Buffer preparation

[0184] 1) 1x Kinase Buffer: 50 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES), pH 7.5, 0.0015% Brij-35, 1 M manganese chloride (MnCl2).

[0185] 2) Stop Buffer: 100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM ethylenediaminetetraacetic acid (EDTA).

[0186] (2) Compound dilution: dilute the compound to the desired concentration with DMSO, 3x gradient dilution of compound solution into 96-well assay plate, a total of 10 concentrations are needed, and another 100 μL DMSO is added to two empty wells of the same 96-well plate as compound blank and enzyme blank controls. After dilution, 40 μL per well is transferred to a 384-well plate, and on this basis, 60 nL per well is added to a 384-well assay plate.

[0187] (3) Kinase reaction: add ATR kinase to 1x kinase buffer solution to configure 2x kinase solution, then take 10 μL solution to add to the above-mentioned 384-well assay plate, incubate at room temperature for 10 min, then add 10 μL of prepared room temperature peptide solution, react at 28°C for a specific time, and finally add 30 μL of termination buffer to stop the kinase reaction.

[0188] (4) Data reading and processing: read and collect data with Caliper, and convert the data to percentage inhibition: percentage inhibition = (maximum value - conversion value) / (maximum value - minimum value) x 100. Curve fitting is performed using analysis software GraphPad Prism to obtain the IC 50 values of the corresponding compounds, as shown in Table 1.

[0189] Table 1 IC 50 values of compounds for ATR kinase

[0190]

[0191]

[0192] Note: ATR 0.1 nM < IC 50 ≤ 10 nM: ***, ATR 10 nM < IC 50 ≤ 100 nM: **, ATR 100 nM < IC 50 ≤ 1000 nM: *.

[0193] As shown in Table 1, all the tested compounds have good inhibition on ATR, with IC 50 values reaching the nanomolar concentration level; among them, compounds I-6, I-9, I-13 to I-16, I-21 to I-22, I-24, I-29, I-31 to I-34, I-41 to I-43, I-45 to I-47, I-49 to I-52, I-62, I-78, I-82, I-86 to I-93, I-95 to I-96, I-100, I-103, I-107, I-109 to I-111, I-113 to I-115, I-117 to I-118, I-120, I-122 to I-124, I-128, I-132 to I-138, and I-140 have IC50 lower than 10 nM, even the IC50 of compounds I-50, I-52 is lower than 1 nM. 50 lower than 1 nM. The above shows that the compounds of the present application have the advantage of developing as ATR inhibitors at the molecular level.

[0194] Example 22: Cell proliferation experiment (test the proliferation inhibition activity of compounds on Lovo and HCT-116 colon cancer cells by CCK-8 method, and take Lovo cell activity test as an example)

[0195] Experimental materials and instruments: F-12K medium (Gibco), fetal bovine serum (Gibco), PBS (Thermo), 0.25% trypsin (Thermo), 96-well cell culture plate (Thermo), CCK-8 (Kaijibio). CO2 cell incubator (Thermo), inverted microscope (Jiangnan Company), Multiskan FC enzyme marker (Thermo), double clean bench (Thermo), blood cell counting board (Shanghai Qiexin biochemical reagent instrument).

[0196] Experimental procedure:

[0197] 1. The Lovo cell strain was cultured to the logarithmic growth phase, the cells were digested and centrifuged. After centrifugation, the cells were resuspended with 1 mL of fresh culture medium, diluted to an appropriate concentration and counted with a cell counter.

[0198] 2. According to the appropriate density, the cells were inoculated into the 96-well plate. Because the evaporation rate of the culture solution in the edge hole of the 96-well plate is faster, the edge 32 holes are not inoculated with cells, and 200 μL of PBS is supplemented.

[0199] 3. The inoculated 96-well plate was placed in a 37℃, 4% CO2 cell incubator for culture. After the cells adhered, drug treatment was performed, 10 concentrations were set for each compound, 2 replicate wells were set for each concentration, and the compound concentration and name were labeled.

[0200] 4. After 72 hours of compound stimulation, the cell state after drug stimulation was observed.

[0201] 5. After 72 hours, the 96-well plate was taken out, CCK-8 solution was added in the dark, and 20 μL of CCK-8 detection reagent was added to each well.

[0202] 6. Place in a 37℃, 4% CO2 incubator for 3-4 hours.

[0203] 7. Take out the 96-well plate, set the enzyme marker program, shake the 96-well plate for 30 s, and then measure the absorbance value at 450 nm. Process the data with Graphpad Prism 8 software, and calculate the IC50 , the results are shown in Tables 2 and 3.

[0204] Table 2 IC inhibition of Lovo cell proliferation by compounds 50 value

[0205] Compound Lovo IC 50 (μM) Compound Lovo IC 50 (μM) Compound Lovo IC 50 (μM) I-1 1.799 I-21 0.726 I-52 1.101 I-3 0.816 I-22 0.930 I-62 0.220 I-6 0.584 I-24 0.255 I-68 0.108 I-9 1.055 I-25 1.303 I-69 0.111 I-10 1.03 I-31 0.743 I-70 0.223 I-11 1.013 I-32 0.461 I-71 0.434 I-12 1.56 I-33 0.726 I-72 0.489 I-13 0.175 I-34 0.690 I-73 0.593 I-14 0.246 I-43 2.433 I-74 0.124 I-15 0.462 I-50 0.461 I-75 0.738 I-16 0.553 I-51 0.648 I-76 1.032 I-77 2.765 I-97 1.11 I-118 0.151 I-78 0.213 I-98 2.70 I-119 1.47 I-79 0.362 I-99 0.884 I-120 0.044 I-80 0.932 I-101 0.648 I-121 0.931 I-81 0.281 I-102 0.635 I-122 0.221 I-82 0.129 I-103 0.177 I-123 0.145 I-83 0.362 I-104 0.819 I-124 0.430 I-84 0.932 I-105 ＞10 I-126 0.388 I-85 0.281 I-106 3.03 I-127 0.214 I-86 0.129 I-107 0.125 I-128 0.582 I-87 0.064 I-108 1.49 I-129 0.441 I-88 0.360 I-109 0.264 I-130 0.268 I-89 0.073 I-110 0.117 I-131 0.035 I-90 0.381 I-111 0.113 I-135 0.178 I-91 0.073 I-112 0.371 I-136 0.163 I-92 0.089 I-113 0.266 I-137 0.377 I-93 0.107 I-114 0.259 I-138 0.370 I-94 0.626 I-115 0.119 I-139 0.694 I-95 0.224 I-116 0.527 I-140 0.060 I-96 0.051 I-117 0.017 AZD-6738 0.598

[0206] Note: AZD-6738 was used as a positive control.

[0207] As shown in Table 2, the tested compounds all had good inhibitory activity against Lovo cell proliferation, IC 50 are less than 5 μM; among them, compounds I-3, I-6, I-13~I-16, I-21~I-22, I-24, I-31~I-34, I-50~I-51, I-62, I -ICs for 68~I-75, I-78~I-81, I-83~I-96, I-99~I-104, I-107, I-109~I-118 and I-120~I-140 50 Less than 1 μM, and compounds I-6, I-13~I-16, I-24, I-32, I-50, I-62, I-68~I-74, I-78~I-79, I-81~I- 83. ICs of I-85~I-93, I-95~I-96, I-103, I-107, I-109~I-118, I-120, I-122~I-138 and I-140 50 It is comparable to the positive control and even better, reaching a concentration level of ten nanomolar.

[0208] Table 3 IC of compounds against HCT-116 cells 50 value

[0209]

[0210]

[0211] Note: BAY-1895344 was used as a positive control.

[0212] As shown in Table 3, the tested compounds all had a good inhibitory effect on HCT-116 cell proliferation, IC 50 All of them reached nanomolar concentrations, among which compounds I-87, I-89, I-92, I-93, I-110 and I-117 had an anti-proliferative effect on HCT-116 cells. 50 The concentrations of the compounds in this application were all less than 100 nM, which was superior to the positive control BAY-1895344. The above results indicate that the compounds in this application have the advantage of inhibiting tumors at the cellular level. Based on the activities at both the molecular and cellular levels, the compounds in this application have highly effective anti-tumor activity.

[0213] Example 23: In vitro liver microsomal metabolic stability study of compounds

[0214] The compounds of the preferred part were subjected to in vitro liver microsomal metabolic stability studies of different species. The test compound was co-incubated with human / rat / mouse liver microsomes (liver microsomes: 0.5 mg·mL -1 Incubation buffer: phosphate buffer (100 mM, pH 7.4); incubation volume: 0.2 mL; incubation time: 0, 5, 15, 30, 60 min; MgCl2: 3 mM; NADPH: 1 mM). The compound concentration in the incubation supernatant was detected, and the data was processed using GraphPad Prism 8.0 software, and the elimination half-life (T 1 / 2 = 0.693 / K) of the test compound in vitro liver microsomal metabolism of different species was obtained, and the results are shown in Table 4. 1 / 2

[0215] Table 4. Elimination half-life (T 1 / 2 ) of compounds in vitro liver microsomal metabolism of different species

[0216]

[0217] As shown in Table 4, the compound I-117 of the present application showed good metabolic stability in vitro liver microsomes of different species.

[0218] In summary, the compounds of the present application showed potent ATR kinase inhibitory activity, and could significantly inhibit the proliferation of cells carrying DNA damage repair defects, while showing good in vitro metabolic stability, which had the advantage of developing as ATR inhibitors.​

Claims

1. A pyrimidocyclic compound, characterized in that: Any of the following compounds: (R)-3-methyl-4-(7-(2-methyl-6-(methylsulfonyl)pyridin-3-yl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-13), Imino(methyl)(6-methyl-5-(4-((R)-3-methylmorpholino)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-7-yl)pyridin-2-yl)-λ 6 -sulfenyl (I-14), (R)-3-methyl-4-(7-(4-methyl-6-(methylsulfonyl)pyridin-3-yl-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-15), Imino(methyl)(4-methyl-5-(4-((R)-3-methylmorpholino)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-7-yl)pyridin-2-yl)-λ 6 -sulfenyl (I-16), (R)-3-methyl-4-(7-(pyrimidin-5-yl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-22), (R)-3-methyl-4-(7-(2-methylsulfonyl)phenyl)-2-(1H-pyrrolo[2,3-b]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-24), (R)-3-methyl-4-(7-(pyridin-4-yl)-2-(1H-pyrrolo[2,3-c]pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)morpholine (I-31), (R)-(3-(4-(3-methylmorpholino)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-2-yl)phenyl)methanol (I-32), (R)-2-(1H-benzo[d]imidazol-1-yl)-4-(3-methylmorpholino)-N-(4-(methylsulfonyl)phenyl)thiophene[3,2-d]pyrimidin-7-amine (I-41), (R)-2-(1H-benzo[d]imidazol-1-yl)-N-(1-methyl-1H-pyrazol-5-yl)-4-(3-methylmorpholino)thienyl[3,2-d]pyrimidin-7-amine (I-42), (R)-((2-(1H-benzo[d]imidazol-1-yl)-4-(3-methylmorpholino)thienyl[3,2-d]pyrimidin-7-yl)imino)dimethyl-λ 6 -sulfenyl (I-43), (R)-4-(2-(1H-benzo[d]imidazol-1-yl)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-4-yl)-3-methylmorpholine (I-49), (R)-1-(4-(3-methylmorpholino)-7-(pyridin-4-yl)thieno[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (I-50), (R)-(2-(1H-benzo[d]imidazol-1-yl)-4-(3-methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)dimethylphosphine oxide (I-52), (R)-N-methyl-1-(4-(3-methylmorpholino)-7-(methylsulfonyl)thieno[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (I-91), (R)-1-(7-(Ethylsulfonyl)-4-(3-methylmorpholino)thieno[3,2-d]pyrimidin-2-yl)-N-methyl-1H-benzo[d]imidazol-2-amine (I-92), (R)-1-(7-(cyclopropylsulfonyl)-4-(3-methylmorpholino)thieno[3,2-d]pyrimidin-2-yl)-N-methyl-1H-benzo[d]imidazol-2-amine (I-93), (R)-1-(7-(isopropylsulfonyl)-4-(3-methylmorpholino)thieno[3,2-d]pyrimidin-2-yl)-N-methyl-1H-benzo[d]imidazol-2-amine (I-95), (R)-N-methyl-1-(4-(3-methylmorpholino)-7-(methylsulfoxy)thieno[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (I-109), (R)-N-methyl-1-(4-(3-methylmorpholino)-7-(methylsulfoximino)thieno[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine (I-110), (R)-(2-(2-(Methylamino)-1H-benzo[d]imidazol-1-yl)-4-(3-methylmorpholino)thieno[3,2-d]pyrimidin-7-yl)dimethylphosphine oxide (I-111).

2. A pharmaceutically acceptable salt of the pyrimidocyclic compound according to claim 1, characterized in that: The salts are salts formed between the compound and hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid or trifluoroacetic acid.

3. A pharmaceutical composition, characterized in that It comprises the pyrimidocyclic compound according to claim 1 or the pharmaceutically acceptable salt according to claim 2 and a pharmaceutically acceptable carrier.

4. Use of the pyrimidocyclic compound according to claim 1 or the pharmaceutically acceptable salt according to claim 2 in the preparation of an ATR inhibitor drug.

5. Use of the pyrimidocyclic compound according to claim 1 or the pharmaceutically acceptable salt according to claim 2 in the preparation of a drug for preventing and / or treating ATR kinase-mediated diseases.

6. The use according to claim 5, characterized in that The ATR kinase-mediated disease is selected from breast cancer, multiple myeloma, bone cancer, bladder cancer, cervical cancer, non-small cell lung cancer, small cell lung cancer, bronchioloalveolar carcinoma, ovarian cancer, esophageal cancer, colorectal cancer, liver cancer, head and neck tumors, kidney cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, testicular cancer, bile duct cancer or leukemia.

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