Heteroaromatic ring compound as well as preparation method and application thereof

Abstract

The invention discloses a heteroaromatic ring compound as well as a preparation method and application thereof. Specifically, the invention relates to compounds of formula (I), and stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope labeled analogues thereof. The invention also relates to a preparation method and application of the compound. # imgabs0 #

Description

Heteroaromatic ring compound and its preparation method and use Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a class of heteroaromatic compounds and their preparation methods and uses. Background Art

[0002] Cancer is one of the leading causes of death in humans. Excessive proliferation of cancer cells is often accompanied by chromosomal instability, as DNA fragments or even entire chromosomes may be duplicated, mutated, or completely lost as cancer cells divide rapidly. This instability is primarily related to cancer invasion. The more unstable the chromosomes, the more likely it is that DNA fragments from these chromosomes will end up in places where they do not originally exist, such as floating in the cytoplasm outside the cell nucleus. Cells recognize these free DNA fragments as evidence of viral invasion, thereby triggering an autoimmune response within the cell, inducing inflammation, and recruiting immune cells to the tumor site to launch an attack.

[0003] The cGAS-STING pathway is an intracellular early warning system involved in this process. When viral or cancer cell chromosomal DNA enters the cytoplasm, cGAS binds to it, forming the second messenger cyclic guanosine monophosphate-adenosine monophosphate (cGAMP). This acts as a warning signal, activating the STING pathway within the cell, leading to the production of type I interferon (IFN) and other co-stimulatory molecules that trigger an immune response. This is a key anti-cancer innate immune pathway, initiating an immune attack against virally infected or cancerous cells. While cancer cells theoretically attract immune cell attacks due to DNA fragments scattered in the cytoplasm, in reality, they can effectively evade immune surveillance, metastasize, and spread. In 2020, scientists at Memorial Sloan Kettering Cancer Center discovered the mechanism that explains this phenomenon: a molecule on the outside of cancer cells destroys the cGAMP signal before it reaches immune cells. This scissor-like protein on the cancer cell surface is ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1), the primary known hydrolase of cGAMP. When cGAMP appears outside the cell, ENPP1 uses its ATPase activity to quickly hydrolyze cGAMP into AMP and GMP, preventing the signal from activating the STING pathway and thus immune cells. AMP also further releases adenosine, an immunosuppressive molecule. Therefore, ENPP1 inhibitors can achieve a "two-pronged approach" in immune regulation.

[0004] Several companies are currently developing small-molecule ENPP1 inhibitors for cancer treatment (Scheme 1). Key international research institutions include Mavupharma (acquired by AbbVie in 2019), Stingray Therapeutics, and Riboscience, LLC. Key domestic research institutions include Nanjing Zhengxiang Pharmaceutical and Betta Pharmaceuticals. Riboscience, LLC's RBS2418 is the most advanced in research, with Phase I enrollment commencing in March 2022. Compounds from other companies are in preclinical development.

[0005] Scheme 1. Structures of major small molecule compounds under development for ENPP1

[0006] RBS2418 is a product under development by Riboscience, LLC. Its structure has not yet been disclosed, and activity data for this compound have not yet been found. The company currently has two patents published on ENPP1 small molecule inhibitors (WO2020210649 and WO2020140001). Its structure is a quinoline and phosphate compound (Scheme 1). Some compounds have in vitro enzyme activity IC 50 The drug is currently enrolling in Phase I clinical trials, with plans to conduct escalation studies of the drug both as a single agent and in combination with the PD-1 antibody perbrolizumab, starting with 100mg BID to 800mg BID, administered orally. Once the dose is determined, efficacy and safety evaluations of both the single agent and combination therapy will be conducted. Phase I clinical trials are expected to be completed in 2024.

[0007] Stingray Therapeutics has published six patents on ENPP1 small molecule inhibition to date, covering two structural types. The main type is quinoline compounds, which differ from Riboscience's compounds mainly in the aminosulfonamide tail portion; the other type is thioether compounds, which have significant structural differences from other molecules under development, and most compounds have an in vitro enzyme activity IC50 of above 100nM.

[0008] MV-626 is an ENPP1 inhibitor developed by MavuPharma, whose structure has not yet been disclosed. In July 2019, AbbVie announced its acquisition of MavuPharma, acquiring its investigational ENPP1 inhibitor, MV-626. Analysis of the company's publicly available patents reveals that its structure primarily consists of variations of the quinoline ring, either bicyclic or tricyclic, with an aminosulfonamide tail. The compound also exhibits nanomolar inhibitory activity against ENPP1 and exhibits favorable PK properties, with 100% oral availability in rats and mice.

[0009] Although some ENPP1 inhibitor small molecules have been disclosed, no ENPP1 inhibitor has been developed and marketed yet. Therefore, there is still an urgent need to develop new compounds with potential for marketing and better efficacy and pharmacokinetic results.

[0010] Summary of the Invention

[0011] The present invention designs a series of compounds having novel structures represented by the general formula, and finds that compounds having such structures exhibit excellent effects and functions, which has positive significance for the development of ENPP1 inhibitors.

[0012] The object of the present invention is to provide a compound with a novel structure as an ENPP1 inhibitor, a preparation method thereof and use thereof in treating diseases mediated by ENPP1.

[0013] The present invention provides a compound of formula (I), and stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs thereof.

[0014] in

[0015] L is selected from -(NR L3 )SO2N(R L1 )(R L2 ),-SO2R L1 、-C(O)OR L1 、-C(O)N(R L1 )(R L2 ),-C(O)NHOR L1 、-P(O)(OR L1 )(OR L2 )、-P(O)(NHR L1 )(OR L2 )、-P(O)(OR L1 )(R L2 )、-P(O)(NHR L1 )(NHR L2 )、-P(O)(NHR L1 )(R L2 )、-P(O)(R L1 )(R L2 )、-P(O)(SR L1 )(SR L2 )、-P(O)(NHR L1 )(SR L2 )、-P(O)(SR L1 )(R L2 ) or -B(OH)2, where R L1 、R L2and R L3 R is independently selected from hydrogen, deuterium, halogen, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, or optionally substituted aryl; or R L1 and R L2 are linked to form an optionally substituted ring structure;

[0016] A is selected from optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, optionally substituted cycloalkylenealkylene, optionally substituted heterocyclylenealkylene, optionally substituted arylenealkylene, or optionally substituted heteroarylenealkylene;

[0017] Z is selected from - (chemical bond), -O-, -S-, -CO-, -SO-, -SO2-, -C(R Z1 )(R Z2 )-、-N(R Z1 )-、-CH2N(R Z1 )-、-N(R Z1 )CH2-、-CON(R Z1 )-、-N(R Z1 )CO-、-SON(R Z1 )-、-SO2N(R Z1 )-、-N(R Z1 )SO-、-N(R Z1 )SO2- or where R Z1 and R Z2 is independently selected from hydrogen, deuterium, halogen, hydroxy, thiol, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;

[0018] Y is selected from -N(R Y )-, -O- or -S-, where R Y is selected from hydrogen, deuterium, optionally substituted alkyl or optionally substituted cycloalkyl;

[0019] n = 0, 1, 2 or 3;

[0020] R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3、-CONR 3 R 4 、-NR 3 COR 4 、-COR 3 、-OCOR 3 、-SONR 3 R 4 、-SO2NR 3 R 4 、-NR 3 SOR 4 、-NR 3 SO2R 4 or -Si(R 3 )(R 4 )(R 5 ),

[0021] R 2 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 or-COR 3 ,

[0022] where R 3 、R 4 and R 5 independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;

[0023] X 1 and X 2 independently selected from N or C(R X ), where R X are independently selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR a 、-SR a 、-NR a R b 、-COOR a 、-CONR a R b 、-NR b COR a 、-COR a 、-OCOR a、-SONR a R b 、-SO2NR a R b 、-NR b SOR a 、-NR b SO2R a or -Si(R a )(R b )(R c ), where R a 、R b and R c R is independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or R 2 With X 1 or X 2 are linked to form an optionally substituted ring structure.

[0024] The present invention also provides a method for treating a disease mediated by ENPP1, comprising administering a compound of formula (I) to a patient in need thereof.

[0025] The present invention also provides use of the compound of formula (I) in preparing a medicament for treating a disease mediated by ENPP1. DETAILED DESCRIPTION

[0026] definition

[0027] As used herein, the term "alkyl", alone or in combination, refers to a straight or branched saturated hydrocarbon chain, including C 1-6 Alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 4-methylpentyl, neopentyl, n-hexyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1,2,2-trimethylpropyl, and the like.

[0028] As used herein, the term "alkenyl", alone or in combination, refers to a straight or branched unsaturated hydrocarbon chain having at least one carbon-carbon double bond, for example, a C-1-1-2-6 carbon atom. 2-6 -alkenyl, such as vinyl, allyl, 1-butenyl, 2-butenyl, isobutenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 1-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, and the like.

[0029] As used herein, the term "alkynyl", alone or in combination, refers to a straight or branched unsaturated hydrocarbon chain having at least one carbon-carbon triple bond, for example a C-1-6 carbon atom. 2-6 -alkynyl, such as ethynyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, 1-hexynyl, 2-hexynyl, 3-hexenyl, and the like.

[0030] As used herein, the term "cycloalkyl", alone or in combination, refers to a saturated non-aromatic cyclic hydrocarbon group, such as a C1-C12-C16 group having 3 to 6 carbon atoms. 3-6 - cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0031] As used herein, the term "heterocyclyl", alone or in combination, refers to a non-aromatic cyclic group containing one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from O, S, and N, such as a 4-6 membered heterocyclyl having 4 to 6 ring atoms or a 4-10 membered heterocyclyl having 4 to 10 ring atoms. Examples of heterocyclyl groups include pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, indolinyl, pyranyl, tetrahydropyran, thiomorpholinyl, tetrahydrothiophenyl, isoxazolidinyl, isothiazolidinyl, oxazolidinyl, thiazolidinyl, azepanyl, benzazepine, and the like.

[0032] As used herein, the term "aryl", alone or in combination, refers to a monocyclic or polycyclic aromatic ring, for example a C1-C2-C10 ring having 6 to 10 carbon atoms. 6-10 Aryl groups such as phenyl, naphthyl, indenyl and the like.

[0033] As used herein, the term "heteroaryl", alone or in combination, refers to a monovalent aromatic heterocyclic monocyclic or polycyclic ring system containing one or more (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, the remaining ring atoms being carbon, e.g., a C 5-6 Heteroaryl, C 5-10 Heteroaryl and C having 5 to 12 ring atoms 5-12 Examples of heteroaryl groups include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, aza Base, diazepine benzothiophene, indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, benzothiophenyl, benzoquinolyl and benzoisoquinolyl.

[0034] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0035] As used herein, the term "hydroxy" refers to an -OH group.

[0036] As used herein, the term "thiol" refers to a -SH group.

[0037] As used herein, the term "amino" refers to a -NH2 group.

[0038] As used herein, the term "cyano" refers to a -CN group.

[0039] As used herein, the term "carbonyl" refers to a -CHO group.

[0040] As used herein, the term "nitro" refers to a -NO2 group.

[0041] As used herein, the term "stereoisomer" refers to a compound composed of the same atoms bonded by the same bonds but having different three-dimensional structures that are not interchangeable. "Stereoisomer" includes enantiomers, diastereomers, and other stereoisomeric forms defined by absolute stereochemistry. The present invention is intended to include all such possible isomers, as well as their racemic and optically pure forms.

[0042] As used herein, the term "geometric isomers" refers to isomeric forms due to carbon-carbon double bonds or other centers of geometric asymmetry.

[0043] As used herein, the term "tautomer" refers to isomeric forms resulting from the interchange of a single bond with an adjacent double bond, along with the concomitant migration of a proton.

[0044] As used herein, the term "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, and sulfuric acid, and salts with organic carboxylic and sulfonic acids such as acetic acid, trifluoroacetic acid, propionic acid, maleic acid, fumaric acid, malic acid, citric acid, tartaric acid, lactic acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, and naphthalenedisulfonic acid; or salts with conventional bases such as alkali metal salts (e.g., sodium and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), ammonium salts derived from ammonia and organic amines (e.g., diethylamine, triethylamine, ethyldiisopropylamine, procaine, dibenzylamine, N-methylmorpholine, dihydroabiethylamine, methylpiperidine, L-arginine, creatine, choline, L-lysine, ethylenediamine, N,N-dibenzylethylenediamine (benzathine), ethanolamine, meglumine, and tromethamine).

[0045] As used herein, the term "prodrug" refers to a compound that undergoes biotransformation before exhibiting its pharmacological effect. Prodrugs can be prepared using well-known methods, such as those described in Burgers Medicinal Chemistry and Drug Development (1995) 172-178, 949-982 (Manfred E. Wolff).

[0046] As used herein, the term "solvate" refers to a complex formed by coordination with a solvent molecule. When the solvent molecule is water, the solvate is a hydrate.

[0047] The compounds described herein may be isotopically labeled, i.e., one or more atoms therein are replaced by atoms having a different atomic mass or mass number. Such isotopically labeled compounds are considered to be within the scope of the present invention. Examples of isotopes that may be incorporated into the compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as, but not limited to, respectively 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I.

[0048] In one aspect, the present invention provides a compound of formula (I), and stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotopically labeled analogs thereof,

[0049] in

[0050] L is selected from -(NR L3 )SO2N(R L1 )(R L2 ),-SO2R L1 、-C(O)OR L1 、-C(O)N(R L1 )(R L2 ),-C(O)NHOR L1 、-P(O)(OR L1 )(OR L2 )、-P(O)(NHR L1 )(OR L2 )、-P(O)(OR L1 )(R L2 )、-P(O)(NHR L1 )(NHR L2 )、-P(O)(NHR L1 )(R L2 )、-P(O)(R L1 )(R L2 )、-P(O)(SR L1 )(SR L2 )、-P(O)(NHR L1 )(SR L2 )、-P(O)(SR L1 )(R L2 ) or -B(OH)2, where R L1 、R L2 and R L3 R is independently selected from hydrogen, deuterium, halogen, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, or optionally substituted aryl; or R L1 and R L2 are linked to form an optionally substituted ring structure;

[0051] A is selected from optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, optionally substituted cycloalkylenealkylene, optionally substituted heterocyclylenealkylene, optionally substituted arylenealkylene, or optionally substituted heteroarylenealkylene;

[0052] Z is selected from -(chemical bond), -O-, -S-, -CO-, -SO-, -SO2-, -C(R Z1 )(R Z2 )-、-N(R Z1 )-、-CH2N(R Z1 )-、-N(R Z1 )CH2-、-CON(R Z1 )-、-N(R Z1 )CO-、-SON(R Z1 )-、-SO2N(R Z1 )-、-N(R Z1 )SO-、-N(R Z1 )SO2- or where R Z1 and R Z2 is independently selected from hydrogen, deuterium, halogen, hydroxy, thiol, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;

[0053] Y is selected from -N(R Y )-, -O- or -S-, where R Y is selected from hydrogen, deuterium, optionally substituted alkyl or optionally substituted cycloalkyl;

[0054] n = 0, 1, 2 or 3;

[0055] R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 、-NR 3 COR 4 、-COR 3 、-OCOR 3 、-SONR 3 R 4 、-SO2NR 3 R 4 、-NR 3 SOR 4 、-NR 3 SO2R 4 or -Si(R 3 )(R 4 )(R 5 ),

[0056] R 2 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR 3 、-SR 3 、-NR 3 R 4 、COOR 3 、-CONR 3 R 4 or-COR 3 ,

[0057] where R 3 、R 4 or R 5 independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;

[0058] X 1 and X 2 independently selected from N or C(R X ), where R X are independently selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR a 、-SR a 、-NR a R b 、-COOR a 、-CONR a R b 、-NR b COR a 、-COR a 、-OCOR a 、-SONR a R b 、-SO2NR a R b 、-NR b SOR a 、-NR b SO2R a or -Si(R a )(R b )(R c ), where R a 、R b and R cR is independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or R 2 With X 1 or X 2 are linked to form an optionally substituted ring structure.

[0059] In some embodiments, R L1 、R L2 and R L3 are independently selected from hydrogen, deuterium, halogen, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group or optionally substituted C 6-10 Aryl.

[0060] In some embodiments, R L1 、R L2 and R L3 independently selected from hydrogen, deuterium, halogen, amino, cyano; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl and aryl; and alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl and aryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclyl (e.g., 4-6 membered heterocyclyl), -OC(O)-alkyl (e.g., -OC(O)-C 1-6 alkyl), -C(O)O-alkyl (e.g. -C(O)OC 1-6 Alkyl) or -OC(O)O-alkyl (e.g. -OC(O)OC 1-6 alkyl).

[0061] In some embodiments, L is selected from: -(NR L3 )SO2N(R L1 )(R L2 ),-SO2R L1 、-C(O)OR L1 、-C(O)NHOR L1 、-P(O)(OR L1 )(OR L2 )、-P(O)(OR L1 )(R L2 )、-P(O)(NHRL1 )(NHR L2 )、-P(O)(NHR L1 )(OR L2 ) or -B(OH)2.

[0062] In some embodiments, R L1 、R L2 and R L3 are independently selected from hydrogen, deuterium, amino, optionally substituted alkyl (e.g., C 1-6 alkyl) and optionally substituted aryl (e.g., C 6-10 aryl).

[0063] In some embodiments, R L1 、R L2 and R L3 independently selected from hydrogen, deuterium, amino, unsubstituted alkyl and phenyl, and alkyl substituted with one or more substituents independently selected from the group consisting of deuterium, -OC(O)-C 1-6 Alkyl, -C(O)OC 1-6 Alkyl, -OC(O)OC 1-6 alkyl.

[0064] In some embodiments, L is selected from: -P(O)(OH)2, -C(O)OH, -C(O)NHOH, -P(O)(OH)(OC2H5), -P(O)(OC2H5)2, -SO2NH2, -NHSO2NH2, -P(O)(OH)(CH3), -P(O)(OCH2OC(O)C(CH3)3)2, -P(O)(NHCH(CH3)C(O)OC2H5)2, -P(O)(NHCH(CH3)C(O)OCH(CH3)2)2, -P(O)(OCH2OC(O)OCH(CH3)2)2, -P(O)(OH)(OCH2OC(O)C(CH3)3), -P(O)(OH)(OC6H5), -P(O)(NHCH(CH3)C(O)OCH(CH3)2)(OC6H5).

[0065] In some embodiments, R L1 and R L2 They are linked to form an optionally substituted 5- or 6-membered ring structure.

[0066] In some embodiments, R L1 and R L2 connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl, optionally substituted 4- to 6-membered heterocyclyl, optionally substituted phenyl (eg, halophenyl), or optionally substituted 5- to 6-membered heteroaryl.

[0067] In some embodiments, R L1 and R L2 connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the following: deuterium; halogen; hydroxyl; thiol; amino; cyano; unsubstituted C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 cycloalkyl, 4-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl; or C substituted by substituents independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, carboxyl, alkyl, cycloalkyl or alkoxy 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 cycloalkyl, 4-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl.

[0068] In some embodiments, L is -P(O)(OR L1 )(OR L2 ), R L1 and R L2 are independently optionally substituted alkyl (e.g., C 1-3 alkyl, such as methyl or ethyl), and R L1 and R L2 The ring structure is connected to form an optionally substituted 5-membered or 6-membered ring structure. In some embodiments, the ring structure is further substituted with phenyl or halophenyl.

[0069] In some embodiments, L is -P(O)(OR L1 )(OR L2 ), R L1 is a methyl group substituted by a halogenated phenyl group, R L2 is ethyl, and R L1 and R L2 They are connected to form a 6-membered ring structure substituted with a halogenated phenyl group.

[0070] In some embodiments, A is selected from optionally substituted C 1-6 Alkylene, optionally substituted C 3-6 Cycloalkylene, optionally substituted 4-10 membered heterocyclylene, optionally substituted C 6-10 Arylene, optionally substituted 5-10 membered heteroarylene, optionally substituted C3-6 Cycloalkylene C 1-3 Alkylene, optionally substituted 4-10 membered heterocyclylene C 1-3 Alkylene, optionally substituted C 6-10 Arylene C 1-3 Alkylene or optionally substituted 5-10 membered heteroarylene C 1-3 Alkylene.

[0071] In some embodiments, A is selected from unsubstituted alkylene, cycloalkylene, heterocyclylene, arylene, heteroarylene, cycloalkylenealkylene, heterocyclylenealkylene, arylenealkylene, or heteroarylenealkylene; and alkylene, cycloalkylene, heterocyclylene, arylene, heteroarylene, cycloalkylenealkylene, heterocyclylenealkylene, arylenealkylene, or heteroarylenealkylene substituted with one or more substituents independently selected from deuterium, halogen, hydroxyl, thiol, amino, cyano, acetyl, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclic group (e.g. 4-6 membered heterocyclic group), -O-alkyl (e.g. -OC 1-6 Alkyl), -NH-alkyl (e.g. -NHC 1-6 Alkyl), -S-alkyl (e.g. -SC 1-6 Alkyl), -O-cycloalkyl (e.g. -OC 1-6 Cycloalkyl), -NH-cycloalkyl (e.g., -NHC 1-6 Cycloalkyl) or -S-cycloalkyl (e.g. -SC 1-6 cycloalkyl).

[0072] In some embodiments, A is selected from methylene, ethylene, propylene, butylene; optionally substituted cyclohexylene, piperidinylene, piperazinylene, diazepanylene, bicyclo[1.1.1]pentylene, phenylene, furanylene, cyclohexylenealkylene, piperidinylenealkylene, piperazinylenealkylene, diazepanylenealkylene, bicyclo[1.1.1]pentylenealkylene, phenylenealkylene, or furanylenealkylene.

[0073] In some embodiments, A is selected from methylene, ethylene, propylene, butylene; unsubstituted or substituted with alkyl (e.g., C 1-6 alkyl, such as methyl) substituted cyclohexylene, piperidinylene, piperazinylene, diazepanylene, bicyclo[1.1.1]pentylene, phenylene or furylene; and unsubstituted or substituted alkyl (e.g. C 1-6 alkyl, such as methyl) substituted cyclohexylene C 1-2 Alkylene, piperidinylene C1-2 Alkylene, piperazinyl C 1-2 Alkylene, diazepanyl C 1-2 Alkylene, bicyclo[1.1.1]pentyl C 1-2 Alkylene, phenylene C 1-2 Alkylene or furylene C 1-2 Alkylene.

[0074] In some embodiments, R Z1 and R Z2 are independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl or an optionally substituted 4- to 6-membered heterocyclic group.

[0075] In some embodiments, R Z1 and R Z2 independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl; and alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from deuterium, halogen, hydroxyl, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 cycloalkyl) or a heterocyclic group (e.g., a 4- to 6-membered heterocyclic group).

[0076] In some embodiments, Z is selected from -(chemical bond), -N(R Z1 )-or-N(R Z1 )CH2-.

[0077] In some embodiments, R Z1 and R Z2 are independently selected from hydrogen, deuterium and optionally substituted alkyl (e.g., C 1-6 alkyl groups, such as methyl groups).

[0078] In some embodiments, Z is selected from -(chemical bond), -NH-, -N(CH3)-, or -NHCH2-.

[0079] In some embodiments, R Y is selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl or optionally substituted C 3-6 Cycloalkyl.

[0080] In some embodiments, RY is selected from hydrogen; deuterium; unsubstituted alkyl or cycloalkyl; and alkyl or cycloalkyl substituted with one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, mercapto, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), -OC(O)-alkyl (e.g. -OC(O)-C 1-6 Alkyl) or -C(O)O-alkyl (e.g. -C(O)OC 1-6 alkyl).

[0081] In some embodiments, R Y Selected from hydrogen, deuterium, unsubstituted or replaced by -OC(O)-C 1-6 Alkyl substituted C 1-6 alkyl.

[0082] In some embodiments, Y is selected from -NH-, -N(CH3)-, -N(CH2OC(O)C(CH3)3)-, -O-, or -S-.

[0083] In some embodiments, R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 Aryl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 、-NR 3 COR 4 、-COR 3 、-OCOR 3 、-SONR 3 R 4 、-SO2NR 3 R 4 、-NR 3 SOR 4 、-NR 3 SO2R 4 or -Si(R 3 )(R 4 )(R 5 ).

[0084] In some embodiments, R 1 is selected from unsubstituted alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl; and alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

[0085] In some embodiments, R 1 selected from alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl substituted with one or more substituents independently selected from the group consisting of deuterium; halogen; hydroxyl; mercapto; amino; cyano; nitro; carboxyl; and alkyl, cycloalkyl or alkoxy substituted with substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, carboxyl, alkyl, cycloalkyl or alkoxy.

[0086] In some embodiments, R 1 Selected from alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl substituted with one or more substituents independently selected from the group consisting of deuterium, fluorine, chlorine, bromine, iodine, hydroxyl, mercapto, amino, cyano, nitro, alkyl, cycloalkyl, alkoxy, alkylamino, carboxyl, haloalkyl, haloalkoxy.

[0087] In some embodiments, R 2 is selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclyl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 or-COR 3 .

[0088] In some embodiments, R 2 is selected from unsubstituted alkyl, alkenyl, cycloalkyl or heterocyclyl; and alkyl, alkenyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

[0089] In some embodiments, R 2selected from alkyl, alkenyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from the group consisting of deuterium; halogen; hydroxyl; mercapto; amino; cyano; nitro; carboxyl; and alkyl, cycloalkyl or alkoxy substituted with substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, carboxyl, alkyl, cycloalkyl or alkoxy.

[0090] In some embodiments, R 2 Selected from alkyl, alkenyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from the group consisting of deuterium, fluorine, chlorine, bromine, iodine, hydroxyl, mercapto, amino, cyano, nitro, alkyl, cycloalkyl, alkoxy, alkylamino, carboxyl, haloalkyl, haloalkoxy.

[0091] In some embodiments, R 3 、R 4 and R 5 are independently selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 aryl or optionally substituted 5-6 membered heteroaryl.

[0092] In some embodiments, R 3 、R 4 and R 5 independently selected from hydrogen; deuterium; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; and alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

[0093] In some embodiments, R 3 、R 4 and R 5 independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl substituted with one or more substituents independently selected from the group consisting of deuterium; halogen; hydroxyl; thiol; amino; cyano; nitro; carboxyl; and alkyl, cycloalkyl or alkoxy substituted with substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, thiol, amino, cyano, nitro, carboxyl, alkyl, cycloalkyl or alkoxy.

[0094] In some embodiments, R 3 、R 4 and R 5independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl substituted with one or more substituents independently selected from the group consisting of deuterium, fluorine, chlorine, bromine, iodine, hydroxyl, mercapto, amino, cyano, nitro, alkyl, cycloalkyl, alkoxy, alkylamino, carboxyl, haloalkyl, haloalkoxy.

[0095] In some embodiments, R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano; unsubstituted or halogen-substituted alkyl, alkenyl, cycloalkyl or aryl (e.g. phenyl); -OR 3 、-NR 3 R 4 、-COOR 3 or -NR 3 COR 4 .

[0096] In some embodiments, R 2 Selected from hydrogen, deuterium, halogen, hydroxyl; alkyl, alkenyl or cycloalkyl which is unsubstituted or substituted by deuterium or halogen; -NR 3 R 4 、-OR 3 or -SR 3 .

[0097] In some embodiments, R 3 and R 4 independently selected from hydrogen; deuterium; alkyl groups which are unsubstituted or substituted with halogen (e.g., C 1-6 alkyl) or cycloalkyl (e.g. C 3-6 cycloalkyl).

[0098] In some embodiments, R 1 Selected from halogen, hydroxy, amino, cyano, -CF3, ethyl, vinyl, cyclopropyl, phenyl substituted with -F, -OCH3, -OCH2CH3, -COOH, -NHCOCH3, -NH-cyclopentyl, -NH(CH3)2.

[0099] In some embodiments, R 2 is selected from halogen, hydroxy, methyl, -CF3, -CHD2, ethyl, propyl, isopropyl, propenyl, cyclopropyl, -NHCH3, -SCH3 or -OCH3.

[0100] In some embodiments, R X are independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, nitro, optionally substituted C 1-6 Alkyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclyl, -OR a 、-SR a 、-NRa R b 、-COOR a 、-CONR a R b 、-NR b COR a 、-COR a 、-OCOR a 、-SONR a R b 、-SO2NR a R b 、-NR b SOR a 、-NR b SO2R a or -Si(R a )(R b )(R c ), where R a 、R b and R c are independently selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 aryl or optionally substituted 5-6 membered heteroaryl; and R X The substituents in the optional substitution are selected from one or more of the following: hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclic group (e.g. 4-6 membered heterocyclic group), aryl group (e.g. C 6-10 aryl) or heteroaryl (e.g., 5-6 membered heteroaryl).

[0101] In some embodiments, X 1 and X 2 are independently selected from N or CH.

[0102] In some embodiments, R 2 With X 1 or X 2 They are linked to form an optionally substituted 5- or 6-membered ring structure.

[0103] In some embodiments, R 2 With X 1 or X 2connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl, optionally substituted 4- to 6-membered heterocyclyl, optionally substituted aryl, or optionally substituted 5- to 6-membered heteroaryl.

[0104] In some embodiments, R 2 With X 1 or X 2 connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the following: deuterium; halogen; hydroxyl; thiol; amino; cyano; unsubstituted C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 cycloalkyl, 4-6 membered heterocyclyl, aryl or 5-6 membered heteroaryl; or C substituted by substituents independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, carboxyl, alkyl, cycloalkyl or alkoxy 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 cycloalkyl, 4-6 membered heterocyclyl, aryl or 5-6 membered heteroaryl.

[0105] In some embodiments, the compound of formula (I), and stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or isotopically labeled analogs thereof are selected from the following compounds:

[0106] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) of the present invention and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or isotope-labeled analogs as an active ingredient. The pharmaceutical composition can be used to prevent and / or treat diseases mediated by ENPP1.

[0107] In some embodiments, the pharmaceutical composition of the present invention may also include additional drugs (such as small molecules, polypeptides, nucleic acids, antibodies, antibody-drug conjugates) for preventing and / or treating cancer or tumors. When the compound of formula (I) of the present invention and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs are administered in combination with additional drugs for preventing and / or treating cancer or tumors, the compound of the present invention or its pharmaceutically acceptable salts, etc. can provide enhanced anti-cancer effects.

[0108] In some embodiments, representative examples of drugs used to treat cancer or tumors include, but are not limited to, cell signaling inhibitors, chlorambucil, melphalan, cyclophosphamide, ifosfamide, busulfan, carmustine, lomustine, streptozotocin, cisplatin, carboplatin, oxaliplatin, dacarbazine, temozolomide, procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine, mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, topotecan, irinotecan, etoposide, trabectedin, dactinomycin, doxorubicin, epirubicin, daunorubicin, mitoxantrone, bleomycin, mitomycin C, ixabepilone, Tamoxifen, flutamide, gonadorelin analogs, megestrol acetate, prednisone, dexamethasone, methylprednisolone, thalidomide, interferon alfa, leucovorin, sirolimus, temsirolimus, everolimus, afatinib, alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crizotinib, dabrafenib, dacomitinib, danuceritid, dasatinib, dovitinib, erlotinib, foretinib, ganetespib, gefitinib, ibrutinib, icotinib, imatinib, iniparib, la Patinib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, saracutinib, saridegib, sorafenib, sunitinib, telatinib, tivan tinib, tivozanib, tofacitinib, trametinib, vandetanib, veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, berentuzumab vedotin, catumaxomab, cetuximab, denosumab, gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, PI3K inhibitors, CSF1R inhibitors, A2A and / or A2B receptor antagonists, IDO inhibitors, anti-PD-1 antibodies, anti-PD-L1 antibodies, LAG3 antibodies, TIM-3 antibodies, TIGIT antibodies, CD47 antibodies, CLAUDIN 18.2 antibodies, anti-CTLA-4 antibodies, or any combination thereof.

[0109] In some embodiments, the pharmaceutical composition of the present invention may further comprise an additional antibody-drug conjugate (ADC) for preventing and / or treating cancer or tumors. When the compound of formula (I) of the present invention and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs are administered in combination with another antibody-drug conjugate for preventing and / or treating cancer or tumors, the compound of the present invention or its pharmaceutically acceptable salt may provide an enhanced anti-cancer effect.

[0110] In some embodiments, representative examples of antibody-drug conjugates for preventing and / or treating cancer or tumors may include, but are not limited to, emtansine trastuzumab, vedicizumab, osartuzumab, vedicizumab, gosartan, delutec-trastuzumab, or any combination thereof.

[0111] In some embodiments, the pharmaceutical composition of the present invention may also include additional therapeutic agents for treating inflammatory diseases, autoimmune diseases and immune-mediated diseases. When the compound of formula (I) of the present invention and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs are administered in combination with additional therapeutic agents for treating inflammatory diseases, autoimmune diseases and immune-mediated diseases, the compound of the present invention or its pharmaceutically acceptable salt, etc. can provide enhanced therapeutic effects.

[0112] In some embodiments, representative examples of therapeutic agents for treating inflammatory diseases, autoimmune diseases, and immune-mediated diseases may include, but are not limited to, steroidal drugs (e.g., prednisone, hydroprednisolone, methylhydroprednisolone, cortisone, hydroxycortisone, betamethasone, dexamethasone, etc.), methotrexate, leflunomide, anti-TNFα agents (e.g., etanercept, infliximab, adalimumab, etc.), calcineurin inhibitors (e.g., tacrolimus, pimecrolimus, etc.), antihistamines (e.g., diphenhydramine, hydroxyzine, loratadine, ebastine, ketotifen, cetirizine, levocetirizine, fexofenadine, etc.), or any combination thereof.

[0113] In yet another aspect, the present invention provides a method for preventing and / or treating a disease mediated by ENPP1, comprising administering the compound of formula (I) or the pharmaceutical composition of the present invention to a patient in need thereof.

[0114] In yet another aspect, the present invention provides use of the compound of formula (I) or pharmaceutical composition of the present invention in the preparation of a medicament for preventing and / or treating a disease mediated by ENPP1.

[0115] In some embodiments, the disease mediated by ENPP1 includes cancer, tumor, inflammatory disease, autoimmune disease, neurodegenerative disease, attention-related disease, or immune-mediated disease.

[0116] In some embodiments, the disease mediated by ENPP1 comprises cancer, tumor, inflammatory disease, autoimmune disease, or immune-mediated disease.

[0117] In some embodiments, the disease mediated by ENPP1 is selected from cancer or tumor. Representative examples of cancer and tumor include, but are not limited to, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis carcinoma, hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, testicular cancer, urinary cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal Tumors, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, leukemia, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder cancer, bronchogenic carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myeloma, liposarcoma, fibrosarcoma, Ewing sarcoma and plasmacytoma.

[0118] In some embodiments, the disease mediated by ENPP1 is selected from inflammatory diseases, autoimmune diseases and immune-mediated diseases, representative examples of which include, but are not limited to, arthritis, rheumatoid arthritis, spondyloarthritis, gouty arthritis, osteoarthritis, juvenile arthritis, other arthritic conditions, lupus, systemic lupus erythematosus (SLE), skin-related diseases, psoriasis, eczema, dermatitis, atopic dermatitis, pain, lung disease, lung inflammation, adult respiratory distress syndrome (ARDS), pulmonary sarcoidosis, chronic inflammatory lung disease, chronic obstructive pulmonary disease, COPD, cardiovascular disease, atherosclerosis, myocardial infarction, congestive heart failure, myocardial ischemia-reperfusion injury, inflammatory bowel disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, asthma, Sjögren's syndrome, autoimmune thyroid disease, urticaria (wheal), multiple sclerosis, scleroderma, organ transplant rejection, xenotransplantation, idiopathic thrombocytopenic purpura (ITP), Parkinson's disease, Alzheimer's disease, diabetes-related diseases, inflammation, pelvic inflammatory disease, allergic rhinitis, allergic bronchitis, and allergic sinusitis.

[0119] Example

[0120] The invention is hereinafter illustrated by the following non-limiting examples.

[0121] The starting materials in the examples of this application are known and can be obtained from commercial suppliers, or can be synthesized according to methods known in the art.

[0122] The structures of the compounds were confirmed by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS) and / or liquid chromatography (HPLC). NMR measurements were performed using a Bruker AVANCE NEO 400 MHz (or 600 MHz); LC-MS measurements were performed using a LCMS WATERS ACQUITY UPLC H-Class PLUS and / or SQD2; and HPLC measurements were performed using a WATERS ACQUITY UPLC and / or Agilent 1260.

[0123] The following abbreviations are used in the examples: AcOH: acetic acid; DCM: dichloromethane; DIBAL-H: diisobutylaluminum hydride; DIEA: N,N-diisopropylethylamine; DMF: N,N-dimethylformamide; DMAP: 4-N,N-dimethylaminopyridine; DMSO: dimethyl sulfoxide; EA: ethyl acetate; HATU: O-(7-nitrobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate; LCMS: liquid chromatography-mass spectrometry; h or hrs: hours; PE: petroleum ether; MeOH: methanol; min: minutes; NCS: N-chlorosuccinimide; rt or RT: room temperature; TFA: trifluoroacetic acid; THF: tetrahydrofuran; TLC: thin layer chromatography; 1N: 1 mol.L -1 (2N: 2mol.L -1 wait).

[0124] The starting materials in the examples of the present invention are known and commercially available, or can be synthesized using or according to methods known in the art.

[0125] Preparation of intermediates:

[0126] Preparation Example 1 (Intermediate 1): Preparation of 7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-2,4-diol:

[0127] Step 1: Preparation of (4-methoxy-2-(4-methoxybenzyl)amino)benzonitrile:

[0128] 2-Fluoro-4-methoxybenzonitrile (CAS: 94610-82-9) (32 g, 0.211 mol, 1.0 equiv) was dissolved in dimethyl sulfoxide (200 mL) at room temperature. 4-Methoxybenzylamine (31.8 g, 0.232 mol, 1.2 equiv) and potassium carbonate (32.0 g, 0.232 mol, 1.2 equiv) were then added. The reaction mixture was heated to 100°C and stirred under argon for 16 hours. The reaction was monitored for completion by LCMS. The reaction mixture was filtered, the filtrate diluted with water (500 mL), and the mixture extracted three times with ethyl acetate (500 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (44 g, 0.164 mol, 77.7% yield) as a yellow solid. LCMS (ESI) [M+H] + =269.1.

[0129] Step 2: Preparation of ethyl 3-amino-6-methoxy-1-(4-methoxybenzyl)-1H-indole-2-carboxylate:

[0130] (4-Methoxy-2-(4-methoxybenzyl)amino)benzonitrile (4 g, 0.011 mol, 1.0 equiv) and ethyl bromoacetate (14.7 g, 0.088 mol, 8.0 equiv) were dissolved in anhydrous N,N-dimethylformamide (80 mL) at room temperature. Potassium tert-butoxide (4.9 g, 0.044 mol, 4 equiv) was then added. The reaction solution was stirred at room temperature for 1 hour under argon. Potassium tert-butoxide (4.9 g, 0.044 mol, 4 equiv) was then added over the next hour, maintaining a light brown color. The mixture was stirred at room temperature for 16 hours, and the reaction was monitored for completion by LCMS. The filtrate was diluted with saturated aqueous ammonium chloride (100 mL), and the mixture was extracted three times with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 6 / 1)) to afford the title compound (1.2 g, 0.003 mol, 30.8% yield) as a yellow oily liquid. LCMS (ESI) [M+H] + =355.0.

[0131] Step 3: Preparation of ethyl 6-methoxy-1-(4-methoxybenzyl)-3-(3-(2,2,2-trichloroacetyl)ureido)-1H-indole-2-carboxylate:

[0132] Ethyl 3-amino-6-methoxy-1-(4-methoxybenzyl)-1H-indole-2-carboxylate (4.5 g, 0.013 mol, 1.0 equivalent) was dissolved in anhydrous tetrahydrofuran (40 mL) at room temperature, followed by the addition of trichloroacetyl isocyanate (2.4 g, 0.013 mol, 1 equivalent). The reaction mixture was stirred at room temperature under argon for 10 minutes. The reaction was monitored for completion by LCMS. The reaction mixture was directly concentrated under reduced pressure to afford the crude title compound (5.8 g, 0.011 mol, 82.4% yield) as a yellow solid. LCMS (ESI) [MH] - =540.2.

[0133] Step 4: Preparation of 7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-2,4-diol:

[0134] Ethyl 6-methoxy-1-(4-methoxybenzyl)-3-(3-(2,2,2-trichloroacetyl)ureido)-1H-indole-2-carboxylate (5.8 g, 0.011 mol, 1.0 equivalent) was dissolved in methanol (60 mL) at room temperature, followed by the addition of sodium hydroxide (2.6 g, 0.066 mol, 6 equivalents). The reaction mixture was heated to 90°C under argon and stirred for 1 hour. The reaction was monitored for completion by LCMS. The reaction mixture was cooled to room temperature, and the pH of the mixture was adjusted to 2 with 1N hydrochloric acid. The resulting suspension was filtered, and the solid was dried to afford the title compound (3.0 g, 0.008 mol, 72.72% yield) as a yellow solid. LCMS (ESI) [M+H] + =352.2.

[0135] Using commercially available substituted 2-fluoro-benzonitrile as the starting material, Intermediate 2, Intermediate 3, Intermediate 4 and Intermediate 5 were prepared according to Steps 1-4 of the preparation method of Preparation Example 1 (Intermediate 1).

[0136] Preparation Example 2 (Intermediate 7): Preparation of 8-bromo-5H-pyrimidin[5,4-b]indole-2,4-diol

[0137] Step 1: Preparation of (4-bromo-2-cyanophenyl)glycinamide:

[0138] 2-Fluoro-5-bromobenzonitrile (30 g, 50 mmol, 1 eq) was added to dimethyl sulfoxide (200 mL), followed by glycinamide hydrochloride (33 g, 300 mmol, 2 eq) and potassium carbonate (51 g, 375 mmol, 2.5 eq). The reaction was heated to 120°C and allowed to react for 16 hours. LCMS monitored the reaction completion. The reaction solution was concentrated under reduced pressure, added with 100 mL of water, filtered, and dried to obtain the title compound (40 g, 149.2 mmol, 99%). LCMS (ESI): [MH] - =252.0,254.0.

[0139] Step 2: Preparation of 3-amino-5-bromo-1H-indole-2-carboxamide:

[0140] (4-Bromo-2-cyanophenyl)glycinamide (20 g, 75.0 mmol) was added to ethanol (150 mL), followed by sodium ethoxide (6.12 g, 90 mmol, 1.2 equivalents), and the reaction was incubated at 82°C for 1 hour. After completion of the reaction, the reaction solution was spin-dried, 100 mL of water was added, filtered, and dried to obtain the title compound (18 g, 67.5 mmol, 90%). LCMS (ESI): [MH] - =251.9,253.9.

[0141] Step 3: Preparation of 8-bromo-5H-pyrimidin[5,4-b]indole-2,4-diol:

[0142] 3-Amino-5-bromo-1H-indole-2-carboxamide (18 g, 67.5 mmol, 1 equivalent) was added to 1,4-dioxane (140 mL), followed by triphosgene (30 g, 101.2 mmol, 1.99 equivalent). The reaction was heated to 120°C and allowed to react for 1 hour. LCMS monitored the reaction completion, and the reaction solution was cooled to room temperature, filtered, and dried to obtain the product 8-bromo-5H-pyrimidin[5,4-b]indole-2,4-diol (15 g, 53.7 mmol, 80%). LCMS (ESI): [MH] - =278.1,280.1.

[0143] Using commercially available substituted 2-fluoro-benzonitrile as the starting material, intermediates 8, 9, 10, 11 and 12 were prepared according to steps 1-3 of the preparation method of Preparation Example 2 (Intermediate 7).

[0144] Preparation Example 3 (Intermediate 13): Preparation of 7,9-difluoro-5H-pyrimido[5,4-b]indole-2,4-diol:

[0145] Step 1: Preparation of 4,6-difluoro-3-nitro-1H-indole-2-carboxylic acid:

[0146] On ice, add concentrated nitric acid (1 ml) to acetic anhydride (9 ml), then add 4,6-difluoro-1H-indole-2-carboxylic acid (CAS: 247564-66-5, 5 g, 25.4 mmol, 1 equivalent). Incubate for two hours on ice until a solid forms. After completion of the reaction, monitored by LCMS, the reaction mixture was filtered to yield the title compound (5.8 g, yield: 94.4%). LCMS (ESI): [MH] - =241.2.

[0147] Step 2: Preparation of methyl 4,6-difluoro-3-nitro-1H-indole-2-carboxylate:

[0148] 4,6-Difluoro-3-nitro-1H-indole-2-carboxylic acid (2 g, 8.26 mmol, 1 eq) was added to methanol (50 mL), followed by thionyl chloride (2 mL, 16.8 mmol, 2.04 eq). The reaction was heated to 80°C and continued for 1 hour. LCMS monitoring confirmed the completion of the reaction. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, neutralized with saturated sodium bicarbonate aqueous solution, and extracted three times with ethyl acetate (20 mL x 3). The mixture was concentrated under reduced pressure to obtain the title compound (600 mg, 28.3%). LCMS (ESI): [MH] - =255.2.

[0149] Step 3: Preparation of 4,6-difluoro-3-nitro-1H-indole-2-carboxamide:

[0150] Methyl 4,6-difluoro-3-nitro-1H-indole-2-carboxylate (2 g, 7.81 mmol, 1 equivalent) was added to 1,4-dioxane (24 mL), followed by aqueous ammonia (24 mL, 0.68 mol, 87.7 equivalents). The reaction was stirred at 100°C for 16 hours. LCMS monitored the completion of the reaction, and the mixture was concentrated under reduced pressure. After dilution with ethyl acetate, the mixture was washed three times with aqueous ammonium chloride solution, and the organic phase was concentrated to obtain the title compound (1.4 g, 74.3%). LCMS (ESI): [MH] - =240.1.

[0151] Step 4: Preparation of 3-amino-4,6-difluoro-1H-indole-2-carboxamide:

[0152] Dissolve 4,6-difluoro-3-nitro-1H-indole-2-carboxamide (250 mg, 1.04 mmol, 1 equivalent) in water (5 mL) and ethanol (5 mL). Add iron powder (289 mg, 5.18 mmol, 5 equivalents) and solid ammonium chloride (554 mg, 10.4 mmol, 10 equivalents). Heat to 80°C and continue stirring for 1 hour. LCMS monitoring indicates completion of the reaction. Cool the reaction solution, concentrate under reduced pressure, add dichloromethane, sonicate for 10 minutes, and filter. The filtrate is extracted three times and concentrated to yield the title compound (150 mg, 68.5%). LCMS (ESI): [M+H] + =212.1.

[0153] Step 5: Preparation of 7,9-difluoro-5H-pyrimido[5,4-b]indole-2,4-diol:

[0154] 3-Amino-4,6-difluoro-1H-indole-2-carboxamide (150 mg, 0.71 mmol, 1 equivalent) was dissolved in 1,4-dioxane (8 mL), and triphosgene (316 mg, 1.07 mmol, 1.5 equivalents) was added. The mixture was heated to 100°C and reacted for 2 hours. LCMS monitored the reaction completion. The reaction solution was cooled and concentrated under reduced pressure to obtain the crude product 7,9-difluoro-5H-pyrimido[5,4-b]indole-2,4-diol (165 mg, 99%), which was used directly in the next reaction. LCMS (ESI): [MH] - =236.1.

[0155] Using commercially available substituted indole-2-carboxylic acid as the starting material, intermediates 14 and 15 were prepared according to steps 1 to 5 of the preparation method of Preparation Example 3 (intermediate 13).

[0156] Preparation Example 4 (Intermediate 16): Preparation of 8-bromo-4-chloro-2-methyl-5H-pyrimidin[5,4-b]indole:

[0157] Step 1: Preparation of ethyl 5-bromo-3-nitro-1H-indole-2-carboxylate:

[0158] Concentrated nitric acid (1 mL) was slowly added to acetic anhydride (9 mL) at 0°C in an ice bath, followed by the addition of ethyl 5-bromo-1H-indole-2-carboxylate (CAS: 16732-70-0, 10 g, 37.3 mmol, 1 equivalent). The reaction mixture was stirred in an ice bath for 1 hour. After completion of the reaction, as monitored by LCMS, the reaction solution was filtered to obtain the title compound (10 g, yield: 85.6%) as a yellow solid. LCMS (ESI): [M+H] + =313.0,315.0.

[0159] Step 2: Preparation of ethyl 3-amino-5-bromo-1H-indole-2-carboxylate:

[0160] Ethyl 5-bromo-3-nitro-1H-indole-2-carboxylate (8.0 g, 25.6 mmol, 1 equivalent) was dissolved in water (40 mL) and ethanol (40 mL). Iron powder (7.1 g, 0.13 mol, 5 equivalents) and ammonium chloride (13.6 g, 0.26 mol, 10 equivalents) were added. The reaction was heated to 80°C and stirred for 1 hour. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, and then extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude title compound (3.8 g, yield: 52.5%) as a yellow solid. LCMS (ESI): [M+H] +=283.2,285.2.

[0161] Step 3: Preparation of 8-bromo-2-methyl-3,5-dihydro-4H-pyrimidin[5,4-b]indol-4-one:

[0162] Ethyl 3-amino-5-bromo-1H-indole-2-carboxylate (2.0 g, 7.06 mmol, 1 equivalent) was dissolved in acetonitrile (5 mL), and dioxane hydrochloride (5 mL, 6 N) was added. The reaction system was sealed and heated to 120°C, where it was stirred for 16 hours. After completion of the reaction, as monitored by LCMS, the reaction solution was diluted with ethyl acetate (50 mL), and the mixture was washed with saturated aqueous sodium bicarbonate (25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (1.5 g, yield: 76.4%) as a yellow solid. LCMS (ESI): [M+H] + =278.2,280.2.

[0163] Step 4: Preparation of 8-bromo-4-chloro-2-methyl-5H-pyrimidin[5,4-b]indole:

[0164] 8-Bromo-2-methyl-3,5-dihydro-4H-pyrimido[5,4-b]indol-4-one (1.5 g, 5.39 mmol, 1 equivalent) was added to phenylphosphonic dichloride (5 mL) at room temperature, and the mixture was stirred at 180°C for 1 hour. After the reaction was completed, the reaction solution was cooled to room temperature and then added to a saturated aqueous sodium bicarbonate solution (100 mL). The mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound (1.2 g, yield: 75.1%) as a yellow solid. LCMS (ESI): [M+H] + =296.1,298.1.

[0165] Commercially available substituted 1H-indole-2-carboxylic acid ethyl ester was used as the starting material and Intermediates 17, 18, 19 and 20 were prepared according to Steps 1-4 of Preparation Example 4 (Intermediate 16).

[0166] Preparation Example 5 (Intermediate 21): Preparation of 8-bromo-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indole-4-ol

[0167] Step 1: Preparation of 8-bromo-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indole-4-ol:

[0168] 5-Bromo-3-nitro-1H-indole-2-carboxamide (the intermediate obtained in Step 2 of Preparation Example 2 (Intermediate 7)) (6 g, 23.7 mmol, 1 equivalent) was dissolved in DCM solvent (120 mL). Triethylamine (7.14 g, 70.9 mmol, 3 equivalents) and trifluoroacetic anhydride (24.8 g, 118.2 mmol, 5 equivalents) were then added to the reaction solution. The reaction mixture was stirred at 25°C for 4 hours. After the reaction was completed, saturated aqueous sodium bicarbonate solution (300 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (120 mL x 3). The organic phase was collected, dried, and concentrated to obtain the title compound (6.05 g, yield: 80.3%). LCMS (ESI): [MH] - =329.9,331.9.

[0169] Using 5-chloroindole-2-carboxylic acid methyl ester as the starting material, the intermediate 3-amino-5-chloro-1H-indole-2-carboxamide was synthesized according to steps 1-3 of the preparation method of Preparation Example 4 (Intermediate 16), and then the intermediate 22 was prepared according to step 1 of the preparation method of Preparation Example 5 (Intermediate 21).

[0170] Intermediate 23 was prepared according to step 1 of the preparation method of Preparation Example 5 (Intermediate 21) using 3-amino-4,6-difluoro-1H-indole-2-carboxamide (the intermediate obtained in step 5 of the synthesis of Preparation Example 3 (Intermediate 8)) as the starting material.

[0171] Preparation Example 6 (Intermediate 24): Preparation of 7,9-difluoro-5H-pyrimido[5,4-b]indol-4-ol:

[0172] Step 1: Preparation of 7,9-difluoro-5H-pyrimido[5,4-b]indol-4-ol:

[0173] 3-Amino-4,6-difluoro-1H-indole-2-carboxamide (the intermediate obtained in Step 5 of Preparation Example 3 (Intermediate 8)) (2 g, 9.47 mmol, 1 equivalent) was dissolved in ethanol (20 mL), and formamidine acetate (2 g, 18.9 mmol, 2 equivalents) was added. The mixture was heated to 100°C and reacted for 2 hours. LCMS monitored the reaction completion. The reaction solution was cooled and concentrated under reduced pressure. The filter cake was washed with ethanol, and the solid was collected and dried to obtain the title compound (1.6 g, 76.4%), which was used directly in the next reaction. LCMS (ESI): [MH] - =220.1.

[0174] Using 5,6-difluoro-1H-indole-2-nonatrimethyl ester as the starting material, the intermediate 3-amino-5,6-difluoro-1H-indole-2-carboxamide (the synthetic starting material of Intermediate 25) was synthesized according to Steps 1-3 of the preparation method of Preparation Example 4 (Intermediate 16).

[0175] Using substituted 3-amino-1H-indole-2-carboxamide as the starting material, intermediates 25 and 26 were prepared according to step 1 of the preparation method of Preparation Example 6 (intermediate 24).

[0176] Preparation Example 7 (Intermediate 27): Preparation of 8-bromobenzo[4,5]thiophene[3,2-d]pyrimidine-2,4(1H,3H)-dione

[0177] Step 1: Preparation of methyl 3-amino-5-bromobenzothiophene-2-carboxylate

[0178] The raw materials 5-bromo-2-fluorobenzonitrile (5 g, 25 mmol, 1 equivalent) and methyl 2-mercaptoacetate (3.18 g, 30 mmol, 1.2 equivalents) were dissolved in DMF (100 mL), cooled to 0°C, and potassium tert-butoxide (5.61 g, 50 mmol, 2 equivalents) was added portionwise. The mixture was stirred at room temperature overnight. The reaction solution was added to water and extracted three times with ethyl acetate (120 mL x 3). The organic phase was washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (4 g, 56%). LCMS (ESI) [M+H] + =286.0,288.0.

[0179] Step 2: Preparation of methyl 5-bromo-3-ureidobenzo[b]thiophene-2-carboxylate

[0180] Methyl 3-amino-5-bromobenzothiophene-2-carboxylate (2.2 g, 7.69 mmol, 1 equivalent) was dissolved in dichloromethane (100 mL), cooled to 0°C, and chlorosulfonyl isocyanate (1.63 g, 11.53 mmol, 1.5 equivalents) was added dropwise. The mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated, and the crude product was dissolved in 6N aqueous hydrochloric acid, heated to 100°C, and stirred for 2 hours. The reaction solution was added to ice water and the pH was adjusted to 8 with 2N sodium hydroxide. A solid precipitated and was filtered to obtain the title compound (2.4 g, 95%) as a brown solid. LCMS (ESI) [M+1] + =329.0,331.0.

[0181] Step 3: Preparation of 8-bromobenzo[4,5]thiophene[3,2-d]pyrimidine-2,4(1H,3H)-dione

[0182] Methyl 5-bromo-3-ureidobenzo[b]thiophene-2-carboxylate (2 g, 6.08 mmol, 1.0 equivalent) was dissolved in methanol (50 mL) and 2N (molar concentration) sodium hydroxide (13.6 mL) was added. The reaction mixture was reacted at 70°C for 3 hours. LCMS confirmed the reaction was complete. The reaction mixture was poured into water, adjusted to pH 3 with 6N (molar concentration) hydrochloric acid, and filtered to obtain the title compound (1.4 g, 78% yield) as a yellow solid. LCMS (ESI) [M+1] + =297.0,299.0.

[0183] Using substituted 2-fluorobenzonitrile and methyl 2-mercaptoacetate as starting materials, intermediate 28 was prepared according to steps 1-3 of the preparation method of Preparation Example 7 (intermediate 27).

[0184] Example 1

[0185] Preparation of (3-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0186] Step 1: Preparation of 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole:

[0187] 7-Methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indole-2,4-diol (1.0 g, 0.003 mol, 1.0 equiv) was dissolved in phosphorus oxychloride (10 mL), followed by the addition of diisopropylethylamine (774 mg, 0.006 mol, 2 equiv). The reaction mixture was heated to 110°C and stirred for 1 hour under argon. The reaction was monitored for completion by LCMS. The reaction mixture was concentrated under reduced pressure, the residue neutralized with saturated aqueous sodium bicarbonate, and the mixture was extracted three times with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, petroleum ether: dichloromethane (V / V = 1 / 1)) to afford the title compound (430 mg, 0.85 mmol, 40.35% yield) as a yellow solid. LCMS: (ESI) [M+H] + =388.1.

[0188] Step 2: Preparation of diethyl (3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0189] Dissolve 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole (200 mg, 0.516 mmol, 1 eq) in acetonitrile (3 mL). Add diethyl (3-aminopropyl)phosphonate (151 mg, 0.774 mmol, 1.5 eq) and triethylamine (156 mg, 1.548 mmol, 3 eq). Heat the reaction mixture to 80°C under argon and stir for 3 hours. LCMS monitoring indicates reaction completion. Add saturated aqueous ammonium chloride (10 mL) to the reaction mixture, and extract the mixture three times with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, dichloromethane:ethyl acetate (V / V = 10 / 1)) to afford the title compound (100 mg, 0.183 mmol, 35.4% yield) as a colorless oil. LCMS (ESI) [M+H] + =547.2.

[0190] Step 3: Preparation of (3-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0191] Diethyl (3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (100 mg, 0.183 mmol, 1 equivalent) was dissolved in dichloromethane (3 mL). Trimethylsilyl bromide (140 mg, 0.915 mmol, 5 equivalents) was then slowly added dropwise to the reaction solution. After addition, the reaction solution was heated to 50°C and stirred for 1 hour. The reaction was monitored for completion by LCMS. The reaction solution was concentrated, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to yield the title compound (7 mg, 0.504 mmol, 48.9% yield) as a white solid. LCMS (ESI) [M+H] + =371.1; 1 H NMR (400 MHz, DMSO-d 6, ppm)δ7.84(d,J=8.8Hz,1H),7.09(s,1H),6.83(d,J=8.4Hz,1H),5.34-5.3 1(m,1H),3.85(s,3H),3.56(br,2H),2.01-1.88(m,2H),1.74-1.69(m,2H).

[0192] Example 2

[0193] Preparation of 5-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)pentanoic acid:

[0194] Step 1: Preparation of ethyl 5-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)pentanoate:

[0195] 2,4-Dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole (100 mg, 0.374 mmol, 1 eq) was dissolved in acetonitrile (2 mL) at room temperature. Ethyl 5-aminopentanoate (163 mg, 1.122 mmol, 3 eq) and triethylamine (113 mg, 1.122 mmol, 3 eq) were then added and stirred at 80°C for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was diluted with saturated aqueous ammonium chloride (50 mL), and the mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by thin-layer chromatography (silica gel, ethyl acetate) to afford the title compound (70 mg, 0.186 mmol, 49.8% yield) as a yellow oil. LCMS: (ESI) [M+H] + =375.2.

[0196] Step 2: Preparation of 5-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)pentanoic acid:

[0197] Ethyl 5-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)pentanoate (70 mg, 0.186 mmol, 1.0 equiv) was dissolved in tetrahydrofuran (1 mL) and water (1 mL) at room temperature. Lithium hydroxide (23 mg, 0.930 mmol, 5 equiv) was then added, and the reaction mixture was stirred at room temperature for half an hour under argon. The reaction was monitored for completion by LCMS. The pH of the reaction mixture was adjusted to 5-6 with 1N hydrochloric acid, and the mixture was extracted three times with ethyl acetate (5 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase HPLC (C18, 10 mmol / L formic acid, acetonitrile) to afford the title compound (5 mg, 0.014 mol, 7.72% yield) as a white solid. LCMS (ESI) [M+H] + =349.0; 1H NMR (400MHz, CD3OD) δ7.95 (d, J=8.8Hz, 1H), 7.00 (d, J=2.0Hz, 1H), 6.86 (dd, J= 8.8Hz,2.0Hz,1H),3.89(s,3H),3.63(br,2H),2.40(br,2H),1.78-1.75(m,4H).

[0198] Example 3

[0199] Similarly, using intermediate 3 as the starting material, Example 3 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI) [M+H] + =419.1. 1 H NMR (400MHz, DMSO-d6, ppm) δ9.35(br,1H),7.89(d,J=8.4Hz,1H),7.79(s,1H),7.28(d,J=8.0Hz,1H),3.54(br,2H),1.94(br,2H),1.76-1.69(m,2H).

[0200] Example 4

[0201] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as the starting material, Example 4 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI) [M+H] + =334.9. 1 H NMR (400MHz, DMSO-d6, ppm) δ7.87(d,J=8.4Hz,1H),7.14(s,1H),7.85(d,J=10.8H z,1H),3.87(s,3H),3.54-3.48(m,2H),2.38(t,J=7.2Hz,2H),2.01-1.98(m,2H).

[0202] Example 5

[0203] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as the starting material, Example 5 was prepared according to steps 2-3 of the preparation method of Example 1. LCMS (ESI) [M+H] + =341.7; 1H NMR (400MHz, DMSO-d6, ppm) δ13.0(br,1H),8.86(br,1H),7.73(d,J=8.8Hz,1H),7.13( br, 2H), 6.96 (s, 1H), 6.75 (d, J = 8.4Hz, 2.0Hz, 1H), 3.90 (s, 3H), 3.60 (d, J = 9.6Hz, 2H).

[0204] Examples 6 and 7

[0205] Preparation of (3-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid (Example 6) and (3-(8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid monoethyl ester (Example 7)

[0206] Step 1: Preparation of 8-bromo-2,4-dichloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole

[0207] 8-Bromo-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-2,4-diol (Intermediate 4) (0.6 g, 1.31 mmol, 1 equivalent) was dissolved in phenylphosphonic dichloride (25 mL) and heated to 150°C under argon with stirring for 3 hours. The reaction was monitored for completion by LCMS. The reaction solution was cooled in an ice bath and then slowly quenched with saturated aqueous sodium bicarbonate solution, adjusting the pH to 6-7. The mixture was extracted three times with ethyl acetate (300 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude title compound (500 mg, 1.15 mmol, 77.5%) as a pale yellow solid. LCMS (ESI) [M+H] + =437.8.

[0208] Step 2: Preparation of diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate

[0209] 8-Bromo-2,4-dichloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole (60 mg, 0.18 mmol, 1 equiv) was dissolved in acetonitrile (10 mL), followed by the addition of diethyl (3-aminopropyl)phosphonate (45 mg, 0.3 mmol, 1.5 equiv) and N,N-diisopropylethylamine (0.06 mL). The reaction was stirred at 80°C for 2 hours. LCMS monitored the reaction completion. The reaction solution was diluted with saturated aqueous ammonium chloride (50 mL), and the mixture was extracted three times with dichloromethane (50 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by thin-layer chromatography (silica gel, ethyl acetate) to afford the title compound (50 mg, 0.084 mmol, 61.8% yield) as a yellow oil. LCMS (ESI) [M+H] + =595.9.

[0210] Step 3: Preparation of diethyl (3-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate

[0211] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.08 mmol, 1 equivalent) was dissolved in trifluoroacetic acid (6 mL), and then 1 drop of anisole (0.02 mL) was added. The reaction solution was stirred at 80°C for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was spin-dried, diluted with ice water (50 mL), and the pH was adjusted to 6-7 with triethylamine. The mixture was extracted three times with dichloromethane (50 mL*3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by thin layer chromatography (silica gel, ethyl acetate) to obtain the title compound (30 mg, 0.064 mmol, 75.3% yield) as a yellow oily liquid. LCMS (ESI) [M+H] + =475.0,477.0.

[0212] Step 4: Preparation of (3-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid (Example 6) and (3-(8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid monoethyl ester (Example 7)

[0213] Diethyl (3-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (30 mg, 0.064 mmol, 1.0 equivalent) was dissolved in dichloromethane (8 mL), and trimethylsilyl bromide (0.5 mL) was added. The reaction solution was heated to 50°C and stirred for 2 hours under argon protection. The reaction was monitored for completion by LCMS. The reaction solution was concentrated under reduced pressure, and the crude product was prepared by reverse phase HPLC (C18 separation column, 10 mmol / L ammonia water, acetonitrile) to give Example 6 (13.94 mg, 0.031 mol, 46.8% yield) and Example 7 (11.97 mg, 0.026 mol, 40.6% yield) as white solids.

[0214] Example 6:

[0215] LCMS (ESI) [M+H] + =419.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.22(d,J=1.6Hz,1H),7.58(dd,J=8.8Hz,1.6Hz,1 H),7.50(d,J=8.8Hz,1H),3.70-3.68(m,2H),2.05-1.99(m,2H),1.85-1.72(m,2H).

[0216] Example 7:

[0217] LCMS (ESI) [M+H] + =447.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.24(d,J=1.6Hz,1H),7.60(dd,J=8.8Hz,2.0Hz,1H),7.51(d,J=8.8Hz,1 H),3.94(q,J=7.2Hz,2H),3.70-3.68(m,2H),2.04-1.96(m,2H),1.78-1.69(m,2H),1.24(t,J=7.2Hz,3H).

[0218] Examples 8 and 9

[0219] Similarly, using intermediate 1 as the starting material, following steps 1-4 of the preparation method of Example 6 and Example 7, Examples 8 and 9 were prepared.

[0220] Example 8

[0221] LCMS (ESI) [M+H] + =427.0; 1H NMR(400MHz,Methanol-d4,ppm)δ7.91(d,J=8.8Hz,1H),7.03(d,J=2.4Hz,1H),6.89(dd,J=8.8Hz,2.4Hz,1H),4.15-4.09(m,4H), 3.90(s,3H),3.75-3.72(m,2H),2.02-1.93(m,4H),1.33(t,J=6.8Hz,6H).

[0222] Example 9

[0223] LCMS (ESI) [M+H] + =399.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.95(d,J=8.8Hz,1H),7.02(d,J=2.0Hz,1H),6.86(dd,J=8.8Hz,2.4Hz,1H),3. 94(q,J=7.2Hz,2H),3.90(s,3H),3.68-3.65(m,2H),2.01-1.99(m,2H),1.79-1.70(m,2H),1.25(t,J=7.2Hz,3H).

[0224] Example 10

[0225] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 10 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI) [M+H] + =321.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.97 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 2.0 Hz, 1H), 6.89 (dd, J = 8. 8Hz, 2.0Hz, 1H), 5.35-5.33 (m, 1H), 3.90 (s, 3H), 2.75 (t, J = 6.0Hz, 2H), 2.19 (t, J = 7.6Hz, 2H).

[0226] Example 11

[0227] Preparation of (3-((7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propylphosphonic acid:

[0228] Step 1: Preparation of diethyl (3-((7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0229] To a solution of diethyl (3-((7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (100 mg, 0.18 mmol) in methanol (5 mL) was added 10% palladium on carbon (20 mg) at room temperature. The reaction mixture was allowed to react at room temperature under a hydrogen atmosphere for 0.5 hour. The reaction was monitored for completion by LCMS. The reaction mixture was filtered and the filtrate was concentrated to obtain the title compound (65 mg, 0.13 mmol, 69%) as a yellow oil. LCMS (ESI) [M+H]+ = 513.3.

[0230] Step 2: Preparation of (3-((7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0231] To a solution of diethyl (3-((7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (20 mg, 0.04 mmol) in acetonitrile (2 mL) was added trimethylsilyl bromide (0.2 mL) at room temperature. The reaction mixture was heated to 70°C for 3 hours. LCMS monitored the completion of the reaction, and the reaction mixture was concentrated. The crude product was purified by reverse phase HPLC (C18, 10 mmol / L ammonia water, acetonitrile) to afford the title compound (1.34 mg, 0.004 mmol, 10%) as a white solid. LCMS (ESI) [M+H] + =337.1; 1 H NMR (400MHz, DMSO-d6, ppm) δ8.24 (s, 1H), 7.85 (dd, J = 8.8Hz, 1H), 7.07 (br, 1H), 6.78 (d ,J=8.0Hz,1H),3.75(br,3H),3.58-3.42(m,2H),2.02-1.94(m,2H),1.80-1.73(m,2H).

[0232] Example 12

[0233] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as the starting material, Example 12 was prepared according to steps 2-3 of the preparation method of Example 1. LCMS (ESI) [M+H]+ =385.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ7.94(d,J=8.8Hz,1H),7.01(d,J=2.4Hz,1H),6.8 5(dd,J=8.8Hz,2.0Hz,1H),3.89(s,3H),3.63(t,J=6.0Hz,2H),1.85-1.66(m,6H).

[0234] Example 13

[0235] Preparation of (3-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)phenyl)phosphonic acid

[0236] Step 1: Preparation of dimethyl (3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)phenyl)phosphonate:

[0237] Dimethyl (3-aminophenyl)phosphonate (47 mg, 0.23 mmol, 1 equivalent), 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole (90 mg, 0.23 mmol, 1 equivalent), tris(dibenzylideneacetone)dipalladium (42 mg, 0.046 mmol, 0.2 equivalent), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (40 mg, 0.069 mmol, 0.3 equivalent) and potassium fluoride (48 mg, 0.82 mmol, 3.5 equivalents) were dissolved in 1,4-dioxane (10 mL), and the reaction solution was heated to 120° C. and reacted for 16 hours. The reaction solution was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to obtain the title compound (30 mg, 0.054 mmol, 24%). LCMS (ESI) [M+H] + =553.2.

[0238] Step 2: Preparation of (3-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)phenyl)phosphonic acid:

[0239] Dissolve dimethyl (3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)phenyl)phosphonate (30 mg, 0.054 mmol) in acetonitrile (5 mL) and add trimethylsilyl chloride (5 mL). Heat the reaction mixture to 90°C for 30 hours. Monitor the reaction by LCMS. The reaction mixture is concentrated under reduced pressure, and the crude product is purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to give the title compound (6 mg, 0.0149 mmol, 28%). LCMS (ESI) [M+H] + =405.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.34(d,J=8.0Hz,1H),8.01(d,J=8.8Hz,1H),7.85(d,J=9.6Hz,1H),8.57(dd ,J=12.0Hz,7.6Hz,1H),7.44-7.39(m,1H),7.11(d,J=1.2Hz,1H),6.89(dd,J=8.8Hz,1.6Hz,1H),3.92(s,3H).

[0240] Example 14

[0241] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as the starting material, Example 14 was prepared according to steps 1-2 of the preparation method of Example 13. LCMS (ESI) [M+H] + =405.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.98 (d, J = 9.2 Hz, 1H), 7.91-7.79 (m, 4H), 7.03 (d, J = 2.4 Hz, 1H), 6.89 (dd, J = 8.8 Hz, 2.0 Hz, 1H), 3.90 (s, 3H).

[0242] Example 15

[0243] Preparation of (2-(1-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)phosphonic acid:

[0244] Step 1: Preparation of dimethyl (E)-(2-(1-benzylpiperidin-4-yl)vinyl)phosphonate:

[0245] 1-Benzylpiperidine-4-carboxaldehyde (2.50 g, 12.3 mmol, 1 equivalent) was dissolved in anhydrous tetrahydrofuran (16 ml), and sodium hydride (295 mg, 12.3 mmol, 1 equivalent) and dimethyl [(dimethoxyphosphonyl)methyl]phosphonate (2.85 g, 12.3 mmol, 1 equivalent) were added. The mixture was stirred at room temperature under argon for two hours. The reaction was monitored for completion by LCMS. After cooling, the reaction was diluted with ethyl acetate (50 mL), and the mixture was extracted three times with saturated aqueous ammonium chloride (25 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude title compound (3.00 g, 9.7 mmol, 78.86%) as a colorless oil. LCMS (ESI): [M+H] + =310.1.

[0246] Step 2: Preparation of dimethyl (2-(piperidin-4-yl)ethyl)phosphonate

[0247] Dissolve dimethyl (E)-(2-(1-benzylpiperidin-4-yl)vinyl)phosphonate (220 mg, 0.71 mmol, 1 eq) in methanol (15 mL) and add palladium on carbon (75 mg, 0.71 mmol, 1 eq). Allow to react at room temperature for 1 hour. LCMS monitored the reaction completion. The reaction solution was filtered and evaporated under reduced pressure to yield the title compound (140 mg, 0.63 mmol, 88.98%) as a light yellow oil. LCMS (ESI): [M+H] + =312.1.

[0248] Step 3: Preparation of dimethyl bis(2-(1-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)phosphonate:

[0249] 2,4-Dichloro-7-methoxy-5-(4-methoxyphenyl)-5H-pyrimido[5,4-b]indole (400 mg, 1.03 mmol, 1 equiv) and dimethyl (2-(piperidin-4-yl)ethyl)phosphonate (456 mg, 2.06 mmol, 2 equiv) were dissolved in acetonitrile (10 mL). N,N-diisopropylethylamine (665 mg, 5.15 mmol, 5 equiv) was added, and the reaction solution was stirred at 100°C for 1 hour. The reaction was monitored for completion by LCMS. The reaction solution was diluted with saturated aqueous ammonium chloride (30 mL), and the mixture was extracted three times with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was isolated and purified by flash chromatography (silica gel, dichloromethane:methanol (v / v = 10 / 1)) to obtain the title compound (450 mg, 76.3%) as a yellow oily solid. LCMS: (ESI) [M+H]+ = 573.2.

[0250] Step 4: Preparation of (2-(1-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)phosphonic acid:

[0251] Dimethyl (2-(1-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)phosphonate (100 mg, 0.18 mmol, 1 eq) was dissolved in dichloromethane (3 mL), and trimethylsilyl bromide (138 mg, 0.9 mmol, 5 eq) was added. The reaction was stirred at 50°C for 1 hour. The reaction was monitored for completion by LCMS. The reaction was quenched with water (2 mL), and the crude mixture was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to give the title compound (25 mg, 0.06 mmol, 32.69%) as a white solid. LCMS: (ESI) [M+H] + =425.2; 1 H NMR(400MHz,Methanol-d4,ppm)δ7.98(d,J=8.8Hz,1H),7.03(d,J=2.0Hz,1H),6.86(dd,J=8.8Hz,2.0Hz,1H) ,4.63-4.59(m,4H),3.90(s,3H),3.21-3.13(m,2H),1.99-1.92(m,2H),1.70-1.59(m,3H),1.39-1.29(m,2H).

[0252] Example 16

[0253] Preparation of 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-sulfonamide:

[0254] Step 1: Preparation of tert-butyl 4-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-carboxylate:

[0255] 2,4-Dichloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indole (116 mg, 0.3 mmol, 1 equiv) was dissolved in acetonitrile (5 mL), followed by the addition of tert-butyl piperazine-1-carboxylate (223 mg, 1.2 mmol, 4 equiv) and N,N-diisopropylethylamine (193 mg, 1.49 mmol, 5 equiv). The reaction mixture was heated to 80°C in a microwave oven under argon protection and stirred for 1 hour. The reaction was monitored for completion by LCMS. The reaction mixture was concentrated, and the crude product was purified by flash chromatography (silica gel, petroleum ether:dichloromethane (V / V=3 / 1)) to afford the title compound (55 mg, 0.1 mmol, 34.07% yield) as a white solid. LCMS: (ESI) [M+H] + =538.4.

[0256] Step 2: Preparation of 1-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine

[0257] Dissolve tert-butyl 4-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-carboxylate (50 mg, 0.09 mmol, 1 equivalent) in trifluoroacetic acid (2.5 mL) and dichloromethane (25 mL). Stir the reaction mixture at room temperature for 2 hours. Saturated aqueous sodium bicarbonate solution was added to the reaction mixture to quench the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude title compound (41 mg, 0.09 mmol) as an orange-brown solid. LCMS: (ESI) [MH] + =438.2.

[0258] Step 3: Preparation of 4-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-sulfonamide:

[0259] 1-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine (41 mg, 0.09 mmol, 1 eq) and sulfonamide (9 mg, 0.09 mmol, 1 eq) were dissolved in 1,4-dioxane (6 mL). Triethylamine (28 mg, 0.28 mmol, 3 eq) was added, and the reaction mixture was heated to 100°C and stirred for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude title compound (58 mg) as an orange-brown solid. LCMS (ESI) [MH] - =515.3.

[0260] Step 4: Preparation of 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-sulfonamide:

[0261] 4-{2-chloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indol-4-yl]piperazine-1-sulfonamide (58 mg, 0.11 mmol, 1 equivalent) was dissolved in trifluoroacetic acid (15 mL), and the mixture was heated to 100°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L formic acid, acetonitrile) to obtain the title compound (1.98 mg, 0.005 mmol, 4.54% yield) as a white solid. LCMS (ESI) [M+H] + =397.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.01(d,J=9.2Hz,1H),7.03(d,J=2.0Hz,1H),6 .89(dd,J=9.2Hz,2.0Hz,1H),4.04-3.99(m,4H),3.91(s,3H),3.33-3.30(m,4H).

[0262] Example 17

[0263] Preparation of 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}benzoic acid:

[0264] Step 1: Preparation of 2,4-dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole

[0265] Dissolve 2,4-dichloro-7-methoxy-5-[(4-methoxyphenyl)methyl]-5H-pyrimidin[5,4-b]indole (93 mg, 0.24 mmol, 1 equivalent) in trifluoroacetic acid (10 mL) and stir in a microwave oven at 90°C under argon protection for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude title compound (107 mg) as a brown oily liquid. LCMS: (ESI) [M+H] + =268.0.

[0266] Step 2: Preparation of methyl 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl]benzoate

[0267] 2,4-Dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole (100 mg, 0.37 mmol, 1 eq) was dissolved in water (1 mL) and 1,4-dioxane (5 mL). Methyl 4-(4,4,5,5-tetramethyl-1,3,2-bis(boronato-2-yl)benzoate (98 mg, 0.37 mmol, 1 eq), potassium carbonate (155 mg, 1.12 mmol, 3 eq), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (55 mg, 0.07 mmol, 0.2 eq) were added. The reaction solution was protected by argon and heated to 80°C with stirring for 1 hour. The reaction solution was cooled to room temperature and diluted with water. The mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (130 mg, 0.35 mmol, 95.53% yield) as a black solid. LCMS: (ESI) [MH] - =366.1.

[0268] Step 3: Preparation of 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}benzoic acid:

[0269] Methyl 4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl]benzoate (130 mg, 0.35 mmol, 1 equivalent) was dissolved in water (5 mL) and tetrahydrofuran (5 mL). Lithium hydroxide monohydrate (148 mg, 3.53 mmol, 10 equivalents) was added, and the reaction mixture was stirred at 30°C for 2 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L formic acid, acetonitrile) to obtain the title compound (5.91 mg, 0.02 mmol, 4.77% yield) as a yellow solid. LCMS (ESI) [MH] -=352.1, [M+H] - =354.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.23(d,J=8.4Hz,2H),8.16(d,J=8.8Hz,1H),8.14 (d,J=8.0Hz,2H),7.11(d,J=2.0Hz,1H),6.97(dd,J=8.8Hz,2.0Hz,1H),3.95(s,3H).

[0270] Example 18

[0271] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as the starting material, Example 18 was prepared according to steps 2-3 of the preparation method of Example 1. LCMS (ESI) [M+H] + =355.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.94 (dd, J = 7.6Hz, 2.0Hz, 1H), 6.99 (s, 1H), 6. 85(dd,J=8.4Hz,1.6Hz,1H),3.90(s,3H),3.90-3.83(m,2H),2.07-2.01(m,2H).

[0272] Example 19

[0273] Preparation of (2-chloro-4-(3-phosphinopropyl)amino)-5H-pyrimidin[5,4-b]indole-7-carboxylic acid:

[0274] Step 1: Preparation of methyl 2,4-dihydroxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-7-carboxylate:

[0275] 7-Bromo-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-2,4-diol (2.0 g, 0.005 mol, 1.0 equiv) and triethylamine (15 ml, 148.2 mmol, 30 equiv) were dissolved in N,N-dimethylformamide (15 mL) and methanol (40 mL). 1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (350 mg, 0.48 mmol, 0.1 equiv) was added to the mixture. The reaction mixture was heated to 125°C under a carbon monoxide atmosphere (3.5 MPa). After 16 hours at this temperature, the reaction mixture was cooled to room temperature, filtered, and the solid was washed with methanol and dried to give the title compound (1.6 g, 4.22 mmol, 84.35%). LCMS (ESI) [MH] - =380.3.

[0276] Step 2: Preparation of methyl 2,4-dichloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-7-carboxylate:

[0277] Methyl 2,4-dihydroxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-7-carboxylate (200 mg, 0.58 mmol, 1.0 equiv) was dissolved in phosphorus oxychloride (3 mL). The reaction solution was heated to 160°C and stirred for 1 hour under argon. The reaction was monitored for completion by LCMS. The reaction solution was concentrated under reduced pressure, the residue was neutralized with saturated aqueous sodium bicarbonate, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, petroleum ether: dichloromethane (V / V = 1 / 1)) to obtain the title compound (60 mg, 0.85 mmol, 40.35% yield) as a yellow solid. LCMS: (ESI) [M+H] + =416.1.

[0278] Step 3: Preparation of methyl 2-chloro-4-((3-(diethoxyphosphino)propyl)amino)-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-7-carboxylate:

[0279] Methyl 2,4-dichloro-5-[(4-methoxybenzyl)methyl]-5H-pyrimido[5,4-b]indole-7-carboxylate (200 mg, 0.48 mmol, 1 equiv) and diethyl 3-aminopropylphosphonate (375 mg, 1.92 mmol, 4 equiv) were dissolved in acetonitrile (5 mL), followed by the addition of diisopropylethylamine (186 mg, 1.44 mmol, 3 equiv). The reaction mixture was heated to 80°C for 16 hours. The reaction mixture was cooled and diluted with saturated aqueous sodium chloride solution (20 mL). The compound was extracted three times with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash preparative plate chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 2 / 1)) to afford the title compound (80 mg, 0.14 mmol, 28.99%) as a yellow, transparent liquid. LCMS (ESI) [M+H] + =575.2.

[0280] Step 4: Preparation of (3-((2-chloro-7-(methoxycarbonyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0281] Dissolve methyl 2-chloro-4-((3-(diethoxyphosphino)propyl)amino)-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole-7-carboxylate (17 mg, 0.030 mmol) in trimethylsilyl chloride (1 mL). Heat the reaction to 90°C for 5 hours. Concentrate under reduced pressure to obtain the crude title compound (10 mg, 0.025 mmol) as a brown solid, which is used directly in the next step.

[0282] Step 5: Preparation of (2-chloro-4-(3-phosphinopropyl)amino)-5H-pyrimidin[5,4-b]indole-7-carboxylic acid:

[0283] Dissolve (3-((2-chloro-7-(methoxycarbonyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid (10 mg, 0.025 mmol) in tetrahydrofuran (3 mL), then add lithium hydroxide (3 mg, 0.075 mmol) and water (0.5 mL). The reaction is allowed to react at room temperature for 16 hours. The reaction solution is directly separated and purified by flash reverse-phase chromatography (C18 column, water, acetonitrile) to obtain the title compound (1.62 mg, 0.004 mmol, 17%) as a white solid. LCMS (ESI) [M+H] + =385.1; 1H NMR (400MHz, Methanol-d4, ppm) δ7.97 (s, 1H), 7.86 (d, J = 8.4Hz, 1H), 7.67 (dd, J =8.4Hz,1.2Hz,1H),3.47(t,J=6.8Hz,2H),1.91-1.85(m,2H),1.70-1.63(m,2H).

[0284] Example 20

[0285] Similarly, using intermediate 4 as the starting material, Example 20 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI) [M+H] + =384.9; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.24(d,J=1.6Hz,1H),7.61(dd,J=8.8Hz,2.0Hz,1H), 7.52(d,J=8.8Hz,1H), 3.69(t,J=6.8Hz,2H), 2.47(t,J=7.2Hz,2H), 2.06-2.01(m,2H).

[0286] Example 21

[0287] Preparation of 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetic acid:

[0288] Step 1: Preparation of methyl 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetate:

[0289] 2,4-Dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole (120 mg, 0.45 mmol, 1 equiv) and methyl 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)phenyl]acetate (87 mg, 0.315 mmol, 0.7 equiv) were dissolved in a mixture of 1,4-dioxane (2.5 mL) and water (0.5 mL) at room temperature. Potassium carbonate (187 mg, 1.35 mmol, 3 equiv) and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) chloride (33 mg, 0.045 mmol, 0.1 equiv) were then added. The mixture was stirred at 80°C under argon for one hour. The reaction was monitored for completion by LCMS. The reaction solution was cooled to room temperature and quenched with water. The mixture was then extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated ammonium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting solid was slurried with petroleum ether:ethyl acetate = 1:1 to afford the target compound, methyl 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetate (90 mg, 0.24 mmol, 52.66%), as a black solid. LCMS: (ESI) [M+H] + =382.0.

[0290] Step 2: Preparation of 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetic acid:

[0291] Methyl 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetate (80 mg, 0.21 mmol, 1 eq) was dissolved in a mixture of water (1.5 mL) and tetrahydrofuran (1.5 mL) at room temperature. Lithium hydroxide (5.02 mg, 0.21 mmol, 1 eq) was added, and the reaction mixture was heated to 50°C and stirred overnight. LCMS monitored the reaction for completion. The reaction mixture was cooled to room temperature, then dilute hydrochloric acid was added to quench the reaction and adjust the solution to neutral. The mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (C18, 10 mmol / L formic acid, acetonitrile) to afford the title compound, 2-(4-{2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl}phenyl)acetic acid (14.36 mg, 0.04 mmol, 18.63%), as a white solid. LCMS (ESI) [M+H] + =368.1; 1H NMR (400MHz, CD3OD, ppm) δ8.15(d,J=8.8Hz,1H),8.07(d,J=8.0Hz,2H),7.58(d,J=8.4H z, 2H), 7.09 (d, J = 2.4Hz, 1H), 6.96 (dd, J = 8.4Hz, 2.4Hz, 1H), 3.94 (s, 3H), 3.77 (s, 2H).

[0292] Example 22

[0293] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 22 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI) [M+H] + =375.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.48(br,1H),7.95(d,J=8.8Hz,1H),6.99(d,J=1.6Hz,1H),6.85(dd,J=8.8Hz,2 .0Hz,1H),4.12-4.07(m,1H),3.89(s,3H),2.32-2.19(m,3H),2.12-2.08(m,2H),1.68-1.60(m,2H),1.45-1.41(m, 2H).

[0294] Example 23

[0295] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 23 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI) [M+H] + =375.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.95 (d, J=8.8Hz, 1H), 7.00 (s, 1H), 6.86 (dd, J=8.8Hz, 2.8Hz ,1H),4.21-4.14(m,1H),3.89(s,3H),2.55-2.42(m,1H),2.20-1.92(m,4H),1.60-1.41(m,4H).

[0296] Example 24

[0297] Preparation of (4-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)benzyl)phosphonic acid:

[0298] Step 1: Preparation of diethyl (4-bromobenzyl)phosphonate:

[0299] 4-Bromo-benzyl bromide (5.0 g, 20.01 mmol, 1 equivalent) was added to triethyl phosphite (50 mL) at room temperature. The reaction mixture was sealed under argon and heated to 80°C with stirring overnight. The reaction was monitored for completion by LCMS. The reaction solution was concentrated under reduced pressure to obtain the crude title compound (5.2 g, 16.93 mmol, 84.62%) as a colorless, transparent liquid. LCMS: (ESI) [M+H] + =306.9.

[0300] Step 2: Preparation of diethyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)benzyl)phosphonate:

[0301] Diethyl (4-bromobenzyl)phosphonate (4 g, 13.02 mmol, 1 equiv) was dissolved in dioxane (20 mL) at room temperature. Diboronic acid pinacol bis(pyrazol)diol (6600 mg, 26.05 mmol, 2 equiv), potassium acetate (2560 mg, 26.05 mmol, 2 equiv), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1450 mg, 1.95 mmol, 0.15 equiv) were added. The reaction solution was protected by argon and heated to 100°C with stirring overnight. The reaction was monitored for completion by LCMS. The reaction solution was cooled to room temperature and diluted with ethyl acetate (200 mL). The mixture was washed with saturated aqueous ammonium chloride (500 mL), and the aqueous phase was extracted three times with ethyl acetate (200 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, ethyl acetate) to give the title compound (3 g, 8.47 mmol, 65.03%) as a colorless liquid. LCMS: (ESI) [M+H] + =355.1.

[0302] Step 3: Preparation of diethyl (4-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)benzyl)phosphonate:

[0303] 2,4-Dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole (100 mg, 0.37 mmol, 1 eq) and diethyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)benzyl)phosphonate (132 mg, 0.37 mmol, 1 eq) were dissolved in a mixture of dioxane (2.5 mL) and water (0.5 mL) at room temperature. Potassium carbonate (155 mg, 1.12 mmol, 3 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (28 mg, 0.04 mmol, 0.1 eq) were then added. The reaction mixture was protected by argon and heated to 80°C with stirring for 2 hours. The reaction was monitored for completion by LCMS. The reaction mixture was cooled to room temperature and diluted with saturated aqueous ammonium chloride (20 mL). The mixture was then extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by preparative thin-layer chromatography (silica gel, dichloromethane:methanol (V / V=10 / 1)) to give the title compound (105 mg, 0.18 mmol, 32.36%) as a yellow solid. LCMS: (ESI) [M+H] + =460.0.

[0304] Step 4: Preparation of (4-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)benzyl)phosphonic acid:

[0305] Diethyl (4-(2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)benzyl)phosphonate (50 mg, 0.11 mmol, 1 equivalent) was dissolved in dichloromethane (1 mL) at room temperature, and trimethylsilyl bromide (85 mg, 0.55 mmol, 5 equivalents) was added. The reaction solution was heated to 50°C and stirred for 2 hours under argon protection. The reaction was monitored for completion by LCMS. The reaction solution was concentrated, and the crude product was prepared by reverse phase HPLC (C18, 10 mmol / L ammonia water, acetonitrile) to give the title compound (5.08 mg, 0.01 mmol, 11.44%) as a white solid. LCMS (ESI) [M+H] + =404.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.12 (dd, J=8.8Hz, 4.0Hz, 1H), 7.99 (d, J=7.2Hz, 2H), 7.59 (d, J=7.6Hz,2H),7.08(s,1H),6.93(dd,J=8.4Hz,2.0Hz,1H),3.94(s,3H),3.13(d,J=21.2Hz,2H).

[0306] Example 25

[0307] Preparation of (3-((8-acetylamino-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0308] Step 1: Preparation of diethyl (3-((8-acetylamino-2-chloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0309] 7-Bromo-1-methyl-2-(trifluoromethyl)quinolin-4(1H)-one (150 mg, 0.25 mmol, 1 equiv) and acetamide (60 mg, 1 mmol, 4 equiv) were suspended in 1,4-dioxane (3 mL). Tris(dibenzylideneacetone)dipalladium(0) (23 mg, 0.03 mmol, 0.1 equiv), 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (29 mg, 0.05 mmol, 0.2 equiv), and cesium carbonate (163 mg, 0.5 mmol, 2 equiv) were added. After purging the system with argon three times, the mixture was heated to 120°C under an argon atmosphere and stirred for 2 hours. The reaction solution was cooled to room temperature, diluted with ethyl acetate, and washed three times with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L formic acid, acetonitrile) to give the title compound (80 mg, 0.14 mmol, 55.75%) as a yellow solid. LCMS: (ESI) [M+H] + =574.1.

[0310] Step 2: Preparation of diethyl (3-((8-acetylamino-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0311] Diethyl (3-((8-acetylamino-2-chloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (80 mg, 0.14 mmol) was dissolved in trifluoroacetic acid (3 mL), and the reaction mixture was stirred at 100°C for 2 hours. The reaction mixture was concentrated under reduced pressure to give the crude title compound (50 mg, 0.11 mmol, 79.05%) as a yellow solid. LCMS: (ESI) [M+H] + =454.1.

[0312] Step 3: Preparation of (3-((8-acetylamino-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0313] Diethyl (3-((8-acetylamino-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.11 mmol, 1 eq) was dissolved in dichloromethane (3 mL). Trimethylsilyl bromide (135 mg, 0.88 mmol, 5 eq) was added, and the reaction mixture was stirred at 50°C for 1 hour. 2 mL of water was added to the reaction mixture, and the aqueous phase was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (3 mg, 0.01 mmol, 4.28%) as a white solid. LCMS: (ESI) [M+H] + =398.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.24 (s, 1H), 7.67 (d, J = 7.2 Hz, 1H), 7.52 (d, J = 8. 8Hz, 1H), 3.69 (t, J = 6.8Hz, 2H), 2.16 (s, 3H), 2.05-1.99 (m, 2H), 1.77-1.67 (m, 2H).

[0314] Example 26

[0315] Preparation of (3-((7-methoxy-2-(methylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0316] Step 1: Preparation of tert-butyl (4-((3-(diethoxyphosphino)propyl)amino)-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-2-yl)(methyl)carbamate:

[0317] Diethyl (3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (105 mg, 0.19 mmol, 1 eq) and tert-butyl methylcarbamate (75 mg, 0.57 mmol, 3 eq) were dissolved in dioxane (9 mL), and tris(dibenzylideneacetone)dipalladium (35 mg, 0.038 mmol, 0.2 eq), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (33 mg, 0.057 mmol, 0.3 eq) and potassium fluoride (40 mg, 0.68 mmol, 4 eq) were added. The reaction system was protected with argon and heated to 120°C for 16 hours. The reaction mixture was concentrated, and the crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to give the title compound (55 mg, 0.086 mmol, yield: 45%) as a brown oil. LCMS (ESI): [M+H] + =642.2.

[0318] Step 2: Preparation of diethyl (3-((7-methoxy-2-(methylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0319] Dissolve tert-butyl (4-((3-(diethoxyphosphino)propyl)amino)-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-2-yl)(methyl)carbamate (55 mg, 0.086 mmol) in trifluoroacetic acid (3 mL), and then heat the reaction mixture under reflux for three hours. The reaction mixture was concentrated to give the crude title compound (30 mg, crude yield: 83%) as a brown oil. LCMS (ESI): [M+H] + =422.2.

[0320] Step 3: Preparation of (3-((7-methoxy-2-(methylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0321] Diethyl (3-((7-methoxy-2-(methylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (30 mg, 0.071 mmol, 1 equivalent) was dissolved in acetonitrile (4 mL) and trimethylsilyl chloride (2 mL) was added. The reaction solution was then heated to 70°C and reacted for 6 hours. The reaction solution was concentrated, and the resulting crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (1.13 mg, yield: 4.3%) as a white solid. LCMS (ESI): [M+H] + =366.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ7.91(d,J=8.8Hz,1H),6.93(s,1H),6.72(d,J=8.8Hz,1H ), 3.86 (s, 3H), 3.60 (t, J = 6.0Hz, 2H), 2.98 (s, 3H), 2.03-1.99 (m, 2H), 1.68-1.63 (m, 2H).

[0322] Example 27

[0323] Preparation of (2-(4-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperazin-1-yl)ethyl)phosphonic acid:

[0324] Step 1: Preparation of diethyl (2-(piperazin-1-yl)ethyl)phosphonate:

[0325] Piperazine (500 mg, 5.8 mmol, 1 eq) was dissolved in ethanol (10 mL), and then diethyl vinylphosphonate (953 mg, 5.8 mmol, 1 eq) was added. The mixture was heated to 80°C and stirred for 2 hours. The ethanol was removed by distillation under reduced pressure to obtain the crude title compound (1.50 g, 5.99 mmol, crude yield: 100%) as a yellow oil. LCMS (ESI): [M+H] + =251.2.

[0326] Step 2: Preparation of diethyl (2-(4-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperazin-1-yl)ethyl)phosphonate:

[0327] Diethyl (2-(piperazin-1-yl)ethyl)phosphonate (406 mg, 1.63 mmol, 1 eq) and 2,4-dichloro-7-methoxy-5H-pyrimido[5,4-b]indole (1.30 g, 4.88 mmol, 3 eq) were dissolved in acetonitrile (5 mL). N,N-diisopropylethylamine (0.5 mL) was added, and the reaction mixture was heated to 90°C and stirred for 2 hours. The reaction mixture was cooled to room temperature, ethyl acetate (5 mL) was added, and the mixture was washed with saturated aqueous ammonium chloride (30 mL). The organic phase was dried and concentrated to give the crude title compound (210 mg, crude yield: 26.8%) as a white solid. LCMS (ESI): [M+H] + =482.2.

[0328] Step 3: Preparation of (2-(4-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperazin-1-yl)ethyl)phosphonic acid:

[0329] Dissolve diethyl (2-(4-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperazin-1-yl)ethyl)phosphonate (50 mg, 0.1 mmol, 1 equivalent) in dichloromethane (2 mL). Add trimethylsilyl bromide (0.5 mL). Heat the reaction mixture to 50°C and stir for 1 hour. Cool the reaction mixture to room temperature and concentrate. The crude product is purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (2 mg, 4.5% yield) as a brown solid. LCMS (ESI): [MH] - =424.1; 1H NMR (400MHz, Methanol-d4, ppm) δ8.00(d,J=8.8Hz,1H),7.03(d,J=2.0Hz,1H),6.88(dd,J=8.8Hz,2.4 Hz, 1H), 3.94 (t, J = 5.2Hz, 4H), 3.91 (s, 3H), 2.85-2.81 (m, 2H), 2.78-2.75 (m, 4H), 1.88-1.77 (m, 2H).

[0330] Example 28

[0331] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 28 was prepared according to steps 2-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =419.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.94(d,J=8.8Hz,1H),7.65(d,J=8.0Hz,2H),7.41(d,J=8 .0Hz, 2H), 7.07 (s, 1H), 6.74 (dd, J = 8.8Hz, 2.0Hz, 1H), 3.88 (s, 3H), 2.91 (d, J = 19.6Hz, 2H).

[0332] Example 29

[0333] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 29 was prepared according to steps 1-4 of the preparation method of Example 16. LCMS (ESI): [M+H] + =411.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.98(d,J=8.8Hz,1H),7.14(d,J=2.0Hz,1H),7.01(dd,J=8.8Hz,2.0Hz,1H),4.31( t,J=5.6Hz,2H),4.24(t,J=6.0Hz,2H),3.94(s,3H),3.71(t,J=5.6Hz,2H),3.46(t,J=6.0Hz,2H),2.14-2.11(m,2H).

[0334] Example 30

[0335] Preparation of (3-((8-amino-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0336] Step 1: Preparation of diethyl (3-((8-amino-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0337] Diethyl (3-((8-acetylamino-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (140 mg, 0.24 mmol, 1 eq) was suspended in methanolic hydrochloric acid (2 mL, 4 M, 8.0 mmol, 33 eq) and the reaction was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and neutralized with saturated aqueous sodium bicarbonate. The mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude title compound (110 mg, yield: 87.5%) as a yellow oil. LCMS (ESI): [M+H] + =532.1.

[0338] Step 2: Preparation of (3-((8-amino-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid

[0339] Diethyl (3-((8-amino-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.094 mmol, 1 equivalent) was dissolved in dichloromethane (1 mL), and trimethylsilyl bromide (0.09 mL) was added. The reaction solution was stirred at 50°C for 1 hour. 1 mL of water was added to the reaction solution, and the aqueous phase was purified by reverse-phase HPLC (C18 column, 10 mmol / L, aqueous ammonia, acetonitrile) to obtain the title compound (1.7 mg, yield: 5.1%) as a white solid. LCMS (ESI): [M+H] + =356.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.40-8.34 (m, 2H), 7.41 (d, J = 8.8Hz, 1H), 7.37 (d, J=2.0Hz,1H),7.02(s,1H),3.64-3.62(m,2H),2.04-2.02(m,2H),1.68-1.64(m,2H).

[0340] Example 31

[0341] Similarly, using intermediate 17 as the starting material, Example 31 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =351.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.01(d,J=8.8Hz,1H),7.08(d,J=2.0Hz,1H),6.84(dd,J=8.8Hz, 2.0Hz,1H),3.91(s,3H),3.65(t,J=6.4Hz,2H),2.59(s,3H),2.08-2.04(m,2H),1.72-1.64(m,2H).

[0342] Example 32

[0343] Preparation of (3-((7-methoxy-2-ethyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0344] Step 1: Preparation of diethyl (3-((7-methoxy-2-vinyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0345] Diethyl (3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (200 mg, 0.47 mmol, 1 equiv) and vinylboronic acid pinacol ester (87 mg, 0.56 mmol, 1.2 equiv) were dissolved in 1,4-dioxane (5 mL) and water (1 mL). To this mixture was added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35 mg, 0.047 mmol, 0.1 equiv) and cesium carbonate (0.46 g, 1.41 mmol, 3 equiv). The reaction system was stirred at 120°C for 4 hours under argon protection. The reaction solution was cooled to room temperature and diluted with saturated aqueous ammonium chloride (20 mL). The mixture was extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 2 / 1)) to give the title compound (100 mg, yield: 50.9%) as a brown solid. LCMS (ESI): [M+H] + =419.1.

[0346] Step 2: Preparation of diethyl (3-((7-methoxy-2-ethyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0347] Diethyl (3-((7-methoxy-2-vinyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (95 mg, 0.23 mmol, 1 equivalent) was dissolved in methanol (1 mL). Palladium on carbon (24 mg, 25 wt%) was then added to the mixture. The reaction was stirred at room temperature under a hydrogen balloon atmosphere for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to provide the crude title compound (90 mg, crude yield: 94.3%) as a brown solid. LCMS (ESI): [M+H] + = 421.2.

[0348] Step 3: Preparation of (3-((7-methoxy-2-ethyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0349] Diethyl (3-((7-methoxy-2-ethyl-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.12 mmol, 1 eq) was dissolved in acetonitrile (1 mL). Trimethylsilyl chloride (65 mg, 0.60 mmol, 5 eq) was then added to the reaction mixture, and the reaction was stirred at 95°C for 2 hours. The organic phase was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to afford the title compound (7 mg, 16% yield) as a yellow solid. LCMS (ESI): [M+H] + =365.13; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.01(d,J=8.8Hz,1H),7.05(d,J=1.6Hz,1H),6.81(dd,J=8.8Hz,2.0Hz,1H),3.89 (s,3H),3.67(t,J=6.4Hz,2H),2.83(q,J=7.6Hz,2H),2.09-2.02(m,2H),1.73-1.64(m,2H),1.35(t,J=7.6Hz,3H).

[0350] Example 33

[0351] Similarly, using intermediate 4 as the starting material, Example 33 was prepared according to steps 1-3 of the preparation method of Example 2. LCMS (ESI): [M+H] + =439.3; 1H NMR (400MHz, DMSO-d6, ppm) δ11.81(brs,1H),8.15(s,1H),7.65-7.62(m,2H),4.57(d,J=13.2Hz,2H),3.15(t, J=12.0Hz,2H),2.29(t,J=7.6Hz,2H),1.86-1.82(m,2H),1.63(br,1H),1.54-1.48(m,2H),1.26-1.23(m,2H).

[0352] Example 34

[0353] Preparation of (3-((2,9-dibromo-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0354] Step 1: Preparation of 9-bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole:

[0355] 9-Bromo-5H-pyrimido[5,4-b]indole-2,4-diol (330 mg, 1.18 mmol, 1 equivalent) was suspended in phenylphosphonyl chloride (20 mL). The mixture was heated to 180°C under an argon atmosphere and stirred for 2 hours. The reaction was monitored for completion by LCMS. After cooling to room temperature, the reaction solution was neutralized with aqueous sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude title compound (310 mg, yield: 83.0%) as a yellow oily liquid. LCMS (ESI): [M+H] + =316.1,318.1,320.1.

[0356] Step 2: Preparation of diethyl (3-((9-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate

[0357] 9-Bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole (310 mg, 0.98 mmol, 1 eq) was dissolved in acetonitrile (3 mL), and diethyl (3-aminopropyl)phosphonate (191 mg, 0.98 mmol, 1 eq) and ethylbis(propan-2-yl)amine (2 mL, 15.5 mmol, 15.8 eq) were added. The reaction was stirred at 100°C for 1 hour. After completion of the reaction, the reaction mixture was added with saturated aqueous ammonium chloride, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase HPLC (C18 column, 10 mmol / L formic acid, acetonitrile) to afford the title compound (300 mg, 64.5% yield) as a yellow solid. LCMS (ESI): [M+H] + =474.9,476.9.

[0358] Step 3: Preparation of (3-((2,9-dibromo-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0359] Diethyl (3-((9-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (150 mg, 0.32 mmol, 1 equivalent) was suspended in dichloromethane (2 mL), and trimethylsilyl bromide (0.3 mL, 1.96 mmol, 6 equivalents) was added. After the system was purged with argon three times, the mixture was heated to 50°C under an argon atmosphere and stirred for 2 hours. After completion of the reaction, monitored by LCMS, 1 mL of water was added, and the aqueous phase was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (30 mg, yield: 20.2%) as a white solid. LCMS (ESI): [MH] - =463.0; 1 H NMR (400MHz, DMSO-d6, ppm) δ11.47(s,1H),7.72-7.68(m,1H),7.45-7.38(m,2H),3.60-3.54(m,2H),1.90-1.82(m,2H),1.72-1.63(m,2H).

[0360] Example 35

[0361] Similarly, using 9-bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole from step 1 of Example 34 as a starting material, Example 35 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI): [M+H]+ =421.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.01 (d, J = 7.6Hz, 1H), 7.50 (d, J = 7.2Hz, 1H), 6. 90(t,J=7.6Hz,1H),3.65(t,J=6.8Hz,2H),2.08-2.01(m,2H),1.67-1.58(m,2H).

[0362] Example 36

[0363] Similarly, using Intermediate 3 as the starting material, Example 36 was prepared according to Steps 1-3 of the preparation method of Example 34. LCMS (ESI) [M+H] + =463.0. 1 H NMR (400MHz, DMSO-d6, ppm) δ9.71 (br, 1H), 7.90 (dd, J = 8.0Hz, 4.0Hz, 1H), 7.79 (s,1H),7.26(d,J=8.4Hz,1H),3.56-3.49(m,2H),1.87(br,2H),1.64(br,2H).

[0364] Example 37

[0365] Preparation of 3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoic acid:

[0366] Step 1: Preparation of methyl 3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoate:

[0367] At room temperature, 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indole (100 mg, 0.26 mmol, 1 eq) and methyl 3-aminobenzoate (59 mg, 0.39 mmol, 1 eq) were dissolved in 1,4-dioxane (3 mL). Cesium fluoride (119 mg, 0.78 mmol, 2 eq), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (30 mg, 0.052 mmol, 0.2 eq), and tris(dibenzylideneacetone)dipalladium (24 mg, 0.026 mmol, 0.1 eq) were added. The reaction mixture was heated to 120°C under argon and stirred for 2 hours. After completion of the reaction, monitored by LCMS, the reaction solution was cooled to room temperature, diluted with ethyl acetate (10 mL), and then washed with saturated aqueous ammonium chloride (10 mL). The aqueous phase was extracted twice with ethyl acetate (10 mL x 2). The organic phases were combined and concentrated. The crude product was purified by silica gel preparative plate chromatography (silica gel, ethyl acetate (100%)) to obtain the title compound (120 mg, yield: 91.8%) as a brown oil. LCMS (ESI): [MH] - =501.4.

[0368] Step 2: Preparation of methyl 3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoate:

[0369] Methyl 3-((2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoate (100 mg, 0.20 mmol, 1 equivalent) was dissolved in trifluoroacetic acid (2 mL) at room temperature. The reaction solution was heated to 100°C and stirred for 2 hours. After completion of the reaction, as monitored by LCMS, the reaction solution was concentrated, the residue was diluted with ethyl acetate (5 mL), and saturated aqueous sodium bicarbonate solution (10 mL) was added to neutralize the solution. The mixture was extracted twice with ethyl acetate (5 mL x 2). The organic phases were combined and concentrated under reduced pressure to give the crude title compound (50 mg, yield: 65.7%) as a yellow solid. LCMS (ESI): [M+H] + =383.3.

[0370] Step 3: Preparation of 3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoic acid:

[0371] Methyl 3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)benzoate (45 mg, 0.12 mmol, 1 equiv) was dissolved in water (1 mL) and tetrahydrofuran (1 mL) at room temperature. Lithium hydroxide (28 mg, 1.18 mmol, 10 equiv) was added, and the reaction mixture was stirred at 50°C for 2 hours. Dilute hydrochloric acid was added to adjust the pH to acidic, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined and concentrated under reduced pressure. The crude product was purified by reverse-phase HPLC (C18, 10 mmol / L, formic acid, acetonitrile) to afford the title compound (1 mg, 2.3% yield) as a white solid. LCMS (ESI): [M+H] + =369.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.34(s,1H),8.19(d,J=6.4Hz,1H),8.02(d,J=8.8Hz,1H),7.76(d,J =8.0Hz, 1H), 7.48 (t, J = 8.0Hz, 1H), 7.08 (d, J = 2.0Hz, 1H), 6.91 (dd, J = 8.8Hz, 2.0Hz, 1H), 3.92 (s, 3H).

[0372] Example 38

[0373] Similarly, using intermediate 4 as the starting material, Example 38 was prepared according to steps 1-3 of the preparation method of Example 15. LCMS (ESI): [M+H] + =475.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.26 (s, 1H), 7.74 (d, J = 9.2 Hz, 1H), 7.64 (d, J = 8.8 Hz,1H),3.53-3.41(m,4H),2.08-2.00(m,2H),1.79-1.60(m,5H),1.46-1.36(m,2H).

[0374] Example 39

[0375] Similarly, using intermediate 4 as the starting material, Example 39 was prepared according to steps 1-3 of the preparation method of Example 2. LCMS (ESI): [M+H] + =425.2; 1H NMR (400MHz, Methanol-d4, ppm) δ8.26(d,J=1.6Hz,1H),7.62(dd,J=8.8Hz,2.0Hz,1H),7.52(d,J=8.8Hz,1H),4.6 9-4.65(m,2H),3.31-3.22(m,2H),2.31(d,J=7.2Hz,2H),2.20-2.15(m,1H),1.98-1.95(m,2H),1.46-1.40(m,2H).

[0376] Example 40

[0377] Similarly, using intermediate 4 as the starting material, Example 40 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): (MH) - =459.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.13(d,J=2.0Hz,1H),7.47(d,J=8.4Hz,1H),7.25(dd,J=8.8Hz,2.0H z,1H),5.39-5.35(m,2H),3.15-3.13(m,2H),2.18-2.14(m,2H),1.53-1.46(m,2H),1.41-1.33(m,3H).

[0378] Example 41

[0379] Preparation of (4-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)cyclohexyl)methyl)phosphonic acid:

[0380] Step 1: Preparation of tert-butyl (4-((dimethoxyphosphonyl)methylene)cyclohexyl)carbamate:

[0381] N-Boc-4-aminocyclohexanone (500 mg, 2.34 mmol, 1 equivalent) was dissolved in tetrahydrofuran (10 mL). Tetramethyl methylenediphosphonate (1.01 g, 4.68 mmol, 2 equivalents) and sodium hydride (187 mg, 4.68 mmol, 2 equivalents) were slowly added sequentially in an ice-water bath. The mixture was stirred at 0°C for 30 minutes. After completion of the reaction, as monitored by LCMS, the reaction was quenched with methanol (10 mL) and water (10 mL). The mixture was concentrated to remove the organic solvent, and the aqueous phase was extracted three times with ethyl acetate (50 mL x 3). The combined organic phases were concentrated to yield the crude title compound (500 mg, crude yield: 66.8%) as a colorless oil. LCMS (ESI): [M+Na]+ =342.2.

[0382] Step 2: Preparation of tert-butyl (4-((dimethoxyphosphonyl)methyl)cyclohexyl)carbamate

[0383] Dissolve tert-butyl (4-((dimethoxyphosphonyl)methylene)cyclohexyl)carbamate (330 mg, 1.03 mmol, 1 equivalent) in methanol (10 mL). Add 10% palladium on carbon (250 mg, 75 wt%), replace the hydrogen atmosphere, and stir at room temperature for 1 hour. Remove the palladium on carbon by filtration, and concentrate the filtrate under reduced pressure to obtain the crude title compound (250 mg, 70.7% yield) as a colorless oil. LCMS (ESI): [M+Na] + =344.4.

[0384] Step 3: Preparation of dimethyl ((4-aminocyclohexyl)methyl)phosphonate

[0385] Dissolve tert-butyl (4-((dimethoxyphosphonyl)methyl)cyclohexyl)carbamate (250 mg, 0.78 mmol, 1 equivalent) in dichloromethane (20 mL). Add trifluoroacetic acid (5 mL) and stir at room temperature for 4 hours. The reaction mixture is concentrated under reduced pressure to afford the crude title compound (150 mg, 87.2% yield) as a brown oil.

[0386] Step 4: Preparation of dimethyl ((4-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)cyclohexyl)methyl)phosphonate

[0387] 8-Bromo-2,4-dichloro-5H-pyrimidin[5,4-b]indole (100 mg, 0.31 mmol, 1 eq) and dimethyl ((4-aminocyclohexyl)methyl)phosphonate (140 mg, 0.62 mmol, 2 eq) were dissolved in acetonitrile (8 mL). Diisopropylethylamine (1 mL) was added and the reaction was heated to 80°C with stirring for 16 hours. The reaction solution was concentrated and then diluted with water (20 mL). The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound (100 mg, crude yield: 63.2%). LCMS (ESI): [M+H] + =5.01.0,503.0.

[0388] Step 5: Preparation of ((4-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)cyclohexyl)methyl)phosphonic acid

[0389] Dimethyl ((4-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)cyclohexyl)methyl)phosphonate (100 mg, 0.20 mmol, 1 equivalent) was dissolved in dichloromethane (10 mL), and trimethylsilyl bromide (3 mL) was added. The reaction was heated to 50°C and stirred for 2 hours. After completion of the reaction, the reaction solution was concentrated, and the crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (10 mg, yield: 10.6%) as a white solid. LCMS (ESI): [M+H] + =473.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.24 (s, 1H), 7.60 (d, J = 7.2Hz, 1H), 7.49 (dd, J = 8.8Hz, 4.0Hz, 1H), 4.13-4.07 (m 1H),2.18-2.10(m,2H),1.89-1.79(m,2H),1.69-1.58(m,2H),1.45-1.23(m,5H).

[0390] Examples 42 and 43

[0391] Preparation of (3-((2-hydroxy-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid (Example 42) and (3-((2-hydroxy-7-methylhydroxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid (Example 43):

[0392] Diethyl (3-((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (300 mg, 0.70 mmol) was dissolved in 6 M HCl / H2O solution (20 mL). The reaction mixture was reacted at 100°C for 16 hours. The reaction solution was concentrated, and the crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the two title compounds:

[0393] Example 42 (6.8 mg, 2.8% yield) was obtained as a white solid. LCMS (ESI): [M+H] + =353.1; 1H NMR (400MHz, Methanol-d4) δ7.79 (d, J = 8.9 Hz, 1H), 6.99 (d, J = 2.1 Hz, 1H), 6.75 (dd, J = 8. 9, 2.2Hz, 1H), 3.89 (s, 3H), 3.63 (t, J = 6.4Hz, 2H), 2.09-1.95 (m, 2H), 1.75-1.61 (m, 2H).

[0394] Example 43 (7.3 mg, 3.1% yield) was obtained as a white solid. LCMS (ESI): [M+H] + =339.1; 1 H NMR (400MHz, Methanol-d4) δ7.47 (d, J = 8.8 Hz, 1H), 6.60-6.49 (m, 2H), 3.59 (t, J = 6.4 Hz, 2H), 2.03-1.93 (m, 2H), 1.73-1.49 (m, 2H).

[0395] Example 44

[0396] Preparation of (4-(((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)methyl)benzyl)phosphonic acid:

[0397] Step 1: Preparation of 2-(4-methylbenzyl)isoindoline-1,3-dione:

[0398] Dissolve 1-(Bromomethyl)-4-methylbenzene (5 g, 26.99 mmol, 1 equivalent) and potassium phthalimide (5 g, 26.99 mmol, 1 equivalent) in N,N-dimethylformamide (20 mL). Stir the reaction mixture at 90°C for 16 hours. Pour the reaction mixture into ice water, stir for half an hour, and filter. The solid is collected and dried to yield the title compound (6 g, 88.4% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.95-7.75(m,4H),7.30-6.98(m,4H),4.72(s,2H),2.26(s,3H).

[0399] Step 2: Preparation of 2-(4-(bromomethyl)benzyl)isoindoline-1,3-dione:

[0400] 2-(4-Methylbenzyl)isoindoline-1,3-dione (6 g, 23.88 mol, 1 equivalent) was dissolved in carbon tetrachloride (50 mL). N-bromosuccinimide (4.3 g, 23.88 mol, 1 equivalent) and dibenzoyl peroxide (578 mg, 2.39 mol, 0.1 equivalent) were added to the reaction mixture. The reaction mixture was refluxed for 1 hour. The solvent of the reaction mixture was dried and purified on a silica gel column (200-300 mesh silica gel, ethyl acetate:petroleum ether = 0-5%) to obtain a crude product. The crude product was slurried with methanol, filtered, and the solid was collected and dried to obtain the title compound (4.3 g, 54.5% yield) as a white solid. LCMS (ESI) [M+23] + =352.1.

[0401] Step 3: Preparation of di-tert-butyl (4-((1,3-dicarbonylisoindolin-2-yl)methyl)benzyl)phosphonate:

[0402] Di-tert-butyl phosphonate (1 g, 5.15 mmol, 1 eq) was dissolved in tetrahydrofuran (20 mL). Sodium hydride (309 mg, 7.73 mmol, 1.5 eq, 60%) was added to the mixture under ice-cooling, and the reaction mixture was stirred at 0°C for 1 hour. 2-(4-(bromomethyl)benzyl)isoindoline-1,3-dione (2.2 g, 6.66 mmol, 1.3 eq) was then added, and the mixture was heated to 70°C and allowed to react for 30 minutes. The reaction mixture was concentrated, and the crude product was purified by flash chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 0-50%) to afford the title compound (700 mg, 30.7% yield) as a colorless oil. LCMS (ESI) [M+23] + =466.2.

[0403] Step 4: Preparation of di-tert-butyl (4-(aminomethyl)benzyl)phosphine ester:

[0404] Dissolve di-tert-butyl (4-((1,3-dicarbonylisoindolin-2-yl)methyl)benzyl)phosphine (700 mg, 1.7 mmol, 1 equivalent) in ethanol (50 mL). Add hydrazine hydrate (425 mg, 8.5 mmol, 5 equivalents) to the reaction mixture, and reflux for 1 hour. Cool the reaction mixture, filter it, and spin-dry the filtrate to obtain the desired product (450 mg, approximately 70% purity) as a colorless oil. LCMS (ESI) [M+1-56-56] + =202.1.

[0405] Step 5: Preparation of di-tert-butyl (4-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)methyl)benzyl)phosphonate:

[0406] 2,4-Dichloro-7-methoxy-5H-pyrimidin[5,4-b]indole (50 mg, 0.19 mmol, 1 eq), di-tert-butyl (4-(aminomethyl)benzyl)phosphonate (59 mg, 0.19 mmol, 1 eq), and triethylamine (38 mg, 0.38 mmol, 2 eq) were added to acetonitrile (5 mL), and the mixture was stirred at 80°C for 16 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (20 mL). The mixture was washed three times with saturated aqueous ammonium chloride (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (30 mg, 29% yield) as a yellow solid. LCMS (ESI) [M+1] + =545.0.

[0407] Step 6: Preparation of ((4-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)methyl)benzyl)phosphonic acid:

[0408] Dissolve di-tert-butyl (4-((2-chloro-7-methoxy-5H-pyrimidin[5,4-b]indol-4-yl)amino)methyl)benzyl)phosphonate (30 mg, 0.05 mmol, 1 eq) in dichloromethane (2 mL) and add trifluoroacetic acid (1 mg, 0.01 mmol, 0.01 eq). Stir at room temperature for 2 hours. Filtration afforded a white solid. The crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to afford the title compound (24 mg, 100%). LCMS (ESI) [M+1] + =433.1; 1 H NMR (400MHz, DMSO-d6) δ2.92(d,J=21.6Hz,2H),3.86(d,J=2.7Hz,3H),4.67(d,J=5.2Hz,2H),6.80(dd,J=2 .3,8.7Hz,1H),7.12(d,J=2.3Hz,1H),7.19-7.41(m,4H),7.87(d,J=8.8Hz,1H),7.94(s,1H),11.01(s,1H).

[0409] Example 45

[0410] Similarly, using the intermediate 2,4-dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole of step 1 of Example 1 as a starting material, Example 45 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI): [M+H] + =383.3; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.00(d,J=8.8Hz,1H),7.76(d,J=8.4Hz,2H),7.32(d,J=8 .8Hz, 2H), 7.05 (d, J = 2.0Hz, 1H), 6.89 (dd, J = 8.8Hz, 2.0Hz, 1H), 3.91 (s, 3H), 3.61 (s, 2H).

[0411] Example 46

[0412] Similarly, using the intermediate of step 1 of Example 34 as the starting material, Example 46 was prepared according to steps 1-2 of the preparation method of Example 2. LCMS (ESI): (MH) - =419.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.55 (d, J = 7.2Hz, 1H), 7.26-7.21 (m, 2H), 3.63 (t, J = 6.8Hz, 2H), 2.07-2.02 (m, 2H), 1.70-1.65 (m, 2H).

[0413] Example 47

[0414] Preparation of (3-((2-chloro-8-(4-fluorophenyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0415] Step 1: Preparation of diethyl (3-((2-chloro-8-(4-fluorophenyl)-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0416] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.084 mmol, 1 equiv) and 4-fluorophenylboronic acid (14 mg, 0.10 mmol, 1.2 equiv) were dissolved in 1,4-dioxane (2 mL) and water (0.4 mL). 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (13 mg, 0.018 mmol, 0.2 equiv) and potassium carbonate (35 mg, 0.25 mmol, 3 equiv) were then added. The mixture was heated to 100°C under nitrogen and stirred for 4 hours. The reaction mixture was diluted with saturated aqueous ammonium chloride (20 mL) and extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 2 / 1)) to give the title compound (50 mg, 97.5% yield) as a brown solid. LCMS (ESI): [M+H] + =611.1.

[0417] Step 2: Preparation of (3-((2-chloro-8-(4-fluorophenyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0418] Diethyl (3-((2-chloro-8-(4-fluorophenyl)-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (45 mg, 0.074 mmol, 1 eq) was dissolved in acetonitrile (1 mL), followed by the addition of trimethylsilyl chloride (40 mg, 0.37 mmol, 5 eq). The reaction was stirred at 95°C for 16 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to afford the title compound (15 mg, 49.3% yield) as a white solid. LCMS (ESI): [M+H] + =435.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.28 (d, J = 1.2Hz, 1H), 7.74-7.64 (m, 4H), 7. 21-7.15(m,2H),3.65(t,J=6.0Hz,2H),2.11-2.01(m,2H),1.75-1.66(m,2H).

[0419] Example 48

[0420] Preparation of (3-((8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)amino)propyl)phosphonic acid:

[0421] Step 1: Preparation of 8-bromo-2,4-dichlorobenzo[4,5]thieno[3,2-d]pyrimidine:

[0422] 8-Bromobenzo[4,5]thiophene[3,2-d]pyrimidine-2,4(1H,3H)-dione (500 mg, 1.68 mmol) was added to phenylphosphonyl dichloride (10 mL), and the reaction mixture was reacted at 180°C for 4 hours. LCMS monitoring confirmed the completion of the reaction. Saturated sodium carbonate solution was added dropwise to the reaction mixture until the pH became weakly alkaline. The mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound (600 mg) as a yellow solid. LCMS: (ESI) [M+H] + =338.8.

[0423] Step 2: Preparation of diethyl (3-(8-bromo-2-chlorobenzo[4,5]thiophene[3,2-d]pyrimidin-4-yl)amino)propyl)phosphonate

[0424] 8-Bromo-2,4-dichlorobenzo[4,5]thieno[3,2-d]pyrimidine (300 mg, 0.90 mmol, 1.0 equiv) was dissolved in MeCN (10 mL). Triethylamine (364 mg, 3.59 mmol, 4.0 equiv) and diethyl (3-aminopropyl)phosphonate (351 mg, 1.80 mmol, 2.0 equiv) were added, and the reaction mixture was allowed to react at 80°C for 18 hours. The reaction mixture was poured into water, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, DCM:MeOH = 30:1) to afford the title compound (380 mg, 86% yield) as a white solid. LCMS (ESI) [M+1]+ = 492.0.

[0425] Step 3: Preparation of (3-((8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)amino)propyl)phosphonic acid

[0426] Diethyl (3-(8-bromo-2-chlorobenzo[4,5]thiophene[3,2-d]pyrimidin-4-yl)amino)propyl)phosphonate (100 mg, 0.20 mmol, 1 equiv) was dissolved in acetonitrile (4 mL) and trimethylsilyl bromide (155 mg, 1.01 mmol, 5.0 equiv) was added. The reaction mixture was reacted at 50°C for 18 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L formic acid, acetonitrile) to obtain the title compound (25 mg, 28% yield) as a white solid. LCMS (ESI): [M+H] + =435.92, [M+3H] + =437.92; 1 H NMR (400MHz, DMSO-d6) δ8.67(t,J=5.3Hz,1H),8.32(d,J=1.8Hz,1H),8.17(d,J=8.6Hz,1H),7.84(d d, J=8.6, 2.0Hz, 1H), 3.55 (dd, J=12.4, 6.5Hz, 2H), 1.84 (dt, J=17.4, 8.9Hz, 2H), 1.70-1.54 (m, 2H).

[0427] Example 49

[0428] Preparation of (3-((8-bromo-2-chlorobenzofuran[3,2-d]pyrimidin-4-yl)amino)propyl)phosphonic acid:

[0429] Step 1: Preparation of 2-(4-bromo-2-cyanophenoxy)acetamide:

[0430] 5-Bromo-2-hydroxybenzonitrile (CAS: 40530-18-5) (5 g, 25.25 mmol, 1.0 eq) was dissolved in DMF (70 mL), potassium carbonate (10.47 g, 75.75 mmol, 3.0 eq) and 2-chloroacetamide (CAS: 79-07-2) (2.83 g, 30.3 mmol, 1.2 eq) were added, and the reaction mixture was stirred at 80°C for 18 hours. After completion of the reaction, the reaction solution was poured into water and filtered. The filter cake was collected to obtain the title compound (5.1 g, 79% yield) as a white solid. LCMS (ESI) [M+1] + =254.9.

[0431] Step 2: Preparation of 3-amino-5-bromobenzofuran-2-carboxamide:

[0432] 2-(4-Bromo-2-cyanophenoxy)acetamide (1 g, 3.92 mmol, 1.0 equiv) was dissolved in EtOH (12 mL) and potassium hydroxide (660 mg, 11.76 mmol, 3.0 equiv) was added. The reaction mixture was allowed to react at 80°C for 2 hours. TLC confirmed the reaction was complete. The reaction mixture was spin-dried and washed with water. The filter cake was collected to yield the title compound (870 mg, 87% yield) as a yellow solid. LCMS (ESI) [M+1] + =255.0.

[0433] Step 3: Preparation of 8-bromobenzofurano[3,2-d]pyrimidine-2,4(1H,3H)-dione:

[0434] 3-Amino-5-bromobenzofuran-2-carboxamide (900 mg, 3.53 mmol, 1.0 equiv) was dissolved in 1,4-dioxane (10 mL), and diphosgene (768 mg, 3.88 mmol, 1.1 equiv) was added. The reaction mixture was reacted at 102°C for 2 hours. LCMS confirmed the reaction was complete. The reaction mixture was poured into water and filtered. The filter cake was collected to yield the title compound (990 mg, 88% yield) as a yellow solid. LCMS (ESI) [M+1] + =281.0.

[0435] Step 4: Using the intermediate of step 3 as a raw material, according to steps 1-3 of the preparation method of Example 48, Example 49 was prepared. LCMS (ESI) [M+1] + =422.0; 1 H NMR (400MHz, DMSO-d6) δ 8.94 (br, 1H), 8.17 (s, 1H), 7.76 (d, J = 20.5Hz, 2H), 3.54 (br, 2H), 1.85 (br, 2H), 1.52 (br, 3H).

[0436] Example 50

[0437] Preparation of (4-(((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)boronic acid:

[0438] 8-Bromo-2,4-dichloro-5H-pyrimidin[5,4-b]indole (100 mg, 0.32 mmol, 1 eq) and 4-aminomethylphenylboronic acid (95 mg, 0.63 mmol, 2 eq) were dissolved in acetonitrile (2 mL), followed by the addition of potassium carbonate (130 mg, 0.95 mmol, 3 eq). The reaction was heated to 70°C under nitrogen and stirred overnight. After completion of the reaction, as monitored by LCMS, the reaction solution was concentrated, and the crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L, formic acid, acetonitrile) to afford the title compound (18.5 mg, 13.4% yield) as a white solid. LCMS (ESI): [M+H] + =431.1,433.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.23 (s, 1H), 7.76 (d, J = 6.8Hz, 2H), 7.58 (dd, J = 8.8Hz, 1.6Hz, 1H), 7.45 (d, J = 8.8Hz, 1H), 7.42 (d, J = 7.2Hz, 2H), 4.58 (s, 2H).

[0439] Example 51

[0440] Similarly, using intermediate 16 as the starting material, Example 51 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): (MH) - =437.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.32 (s, 1H), 7.56 (d, J = 8.8Hz, 1H), 7.50 (d, J = 8.8Hz, 1H), 4.63-4. 57(m,2H),3.23-3.12(m,2H),2.58(s,3H),2.21-2.17(m,3H),2.05-2.01(m,2H),1.54-1.48(m,2H).

[0441] Example 52

[0442] Similarly, using intermediate 16 as the starting material, Example 52 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =399.1; 1H NMR (400MHz, Methanol-d4, ppm) δ8.27 (s, 1H), 7.51 (br, 2H), 3.65 (t, J = 6.0Hz, 2H), 2.58 (s, 3H), 2.09-2.02 (m, 2H), 1.70-1.65 (m, 2H).

[0443] Example 53

[0444] Similarly, using intermediate 20 as the starting material, Example 53 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =467.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.34(d,J=1.6Hz,1H),7.54(dd,J=8.8Hz,1.6Hz,1H),7.48(d,J=8.8Hz,1H),4.60-4.56(m,2 H),3.15-3.12(m,2H),2.20-2.13(m,4H),1.51(d,J=6.4Hz,1H),1.47(d,J=6.4Hz,1H),1.34-1.28(m,2H),1.13-1.07(m,4H).

[0445] Example 54

[0446] Similarly, using intermediate 20 as the starting material, Example 54 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): (MH) - =425.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.30(d,J=1.6Hz,1H),7.52-7.45(m,2H),3.63(t,J=6.0Hz,2H) ,2.13-2.10(m,1H),2.04-1.98(m,2H),1.66-1.60(m,2H),1.13-1.10(m,2H),0.92-0.88(m,2H).

[0447] Example 55

[0448] Preparation of (3-((2-chloro-8-cyano-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0449] Step 1: Preparation of diethyl (3-((2-chloro-8-cyano-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0450] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.084 mmol, 1 equiv) and zinc cyanide (20 mg, 0.17 mmol, 2 equiv) were dissolved in N,N-dimethylformamide (1 mL). Palladium acetate (4 mg, 0.018 mmol, 0.2 equiv) and 1,1'-bis(diphenylphosphino)ferrocene (19 mg, 0.034 mmol, 0.4 equiv) were then added to the mixture. The reaction solution was protected with nitrogen and heated to 120°C in a microwave oven for 1 hour. The reaction solution was diluted with saturated aqueous ammonium chloride (20 mL) and extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by thin-layer preparative plate chromatography (silica gel, petroleum ether:ethyl acetate (V / V=3 / 1)) to afford the title compound (25 mg, yield: 55.0%) as a brown solid. LCMS (ESI): [M+H] + =542.0.

[0451] Step 2: Preparation of (3-((2-chloro-8-cyano-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0452] Diethyl (3-((2-chloro-8-cyano-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (20 mg, 0.04 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl chloride (0.2 mL) was then added to the mixture, and the reaction mixture was stirred at 95°C for 16 hours. After completion of the reaction, which was monitored by LCMS, the reaction mixture was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to yield the title compound (5 mg, 34.2% yield) as a white solid. LCMS (ESI): [MH] - =364.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.44 (s, 1H), 7.75-7.68 (m, 2H), 3.69 (t, J = 6.4Hz, 2H), 2.13-2.02 (m, 2H), 1.83-1.74 (m, 2H).

[0453] Example 56

[0454] Preparation of (3-((2-chloro-8-(cyclopentylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0455] Step 1: Preparation of diethyl (3-((2-chloro-8-(cyclopentylamino)-5-(4-methoxybenzyl)-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0456] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (100 mg, 0.17 mmol, 1 equiv) and cyclopentylamine (29 mg, 0.34 mmol, 2 equiv) were dissolved in N,N-dimethylformamide (1 mL). Methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (27 mg, 0.034 mmol, 0.2 equiv) and cesium carbonate (164 mg, 0.5 mmol, 3 equiv) were added to the mixture. The reaction solution was heated to 120°C in a microwave under nitrogen protection for 1 hour. The reaction mixture was diluted with saturated aqueous ammonium chloride (30 mL), and the mixture was extracted three times with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative thin-layer plate chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 3 / 1)) to afford the title compound (50 mg, yield: 49.7%) as a brown solid. LCMS: (ESI) [M+H] + =600.3.

[0457] Step 2: Preparation of (3-((2-chloro-8-(cyclopentylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0458] Diethyl (3-((2-chloro-8-(cyclopentylamino)-5-(4-methoxybenzyl)-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.083 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl chloride (0.2 mL) was then added to the mixture, and the reaction was stirred at 95°C for 16 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to yield the title compound (4 mg, 0.01 mmol, 11.33%) as a white solid. LCMS: (ESI) [M+H] +=424.2; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.37(d,J=8.8Hz,1H),7.33(d,J=2.0Hz,1H),7.01(dd,J=8.8Hz,2.0Hz,1H),3.9 0-3.82(m,1H),3.62(t,J=6.0Hz,2H),2.11-2.01(m,4H),1.79-1.74(m,2H),1.73-1.63(m,4H),1.59-1.52(m,2H).

[0459] Example 57

[0460] Preparation of ((1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0461] Step 1: Preparation of diethyl ((1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphonate:

[0462] 8-Bromo-2,4-dichlorobenzo[4,5]thieno[3,2-d]pyrimidine (300 mg, 0.90 mmol, 1 equiv) was dissolved in MeCN (10 mL), and TEA (363 mg, 3.59 mmol, 4 equiv) and diethyl (piperidin-4-ylmethyl)phosphonate hydrochloride (488 mg, 1.80 mmol, 2.0 equiv) were added. The reaction mixture was allowed to react at 25°C for 1 hour. LCMS monitored the reaction completion. The reaction solution was poured into water and extracted with E. The EA phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then purified by flash chromatography (200-300 mesh silica gel, DCM:MeOH = 30:1) to afford the title compound (420 mg, 88% yield) as a white solid. LCMS (ESI) [M+1]+ = 532.1.

[0463] Step 2: Preparation of ((1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0464] Diethyl ((1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphonate (370 mg, 0.69 mmol, 1 eq) was dissolved in MeCN (15 mL) and TMSBr (532 mg, 3.45 mmol, 5.0 eq) was added. The reaction mixture was reacted at 50°C for 16 hours. LCMS monitored the reaction completion. The reaction solution was then dried and then purified by acetonitrile and water (0.1% TFA) to yield the title compound (30 mg, 9% yield) as a white solid. LCMS (ESI) [M+1H] + =475.95, [M+3H] + =477.95; 1 H NMR (400MHz, DMSO-d6) δ8.32(d,J=2.0Hz,1H),8.10(d,J=8.6Hz,1H),7.85(dd,J=8.6,2.1Hz,1H),4.60(d,J=13.4Hz,2H), 3.30(d,J=12.8Hz,2H),2.09-2.00(m,3H),1.54(dd,J=18.2,6.0Hz,2H),1.34(q,J=12.4Hz,2H).

[0465] Example 58

[0466] Similarly, using Intermediate 2 as the starting material, Example 58 was prepared according to Steps 1-2 of the preparation method of Example 57. LCMS (ESI): [M+H] + =411.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.57(d,J=2.4Hz,1H),7.47(d,J=9.2Hz,1H),7.16(dd,J=8.8Hz,2.4Hz,1H),4.65-4.61(m, 2H),3.87(s,3H),3.24(t,J=13.2Hz,2H),2.15-2.11(m,3H),1.68(d,J=6.0Hz,1H),1.63(d,J=6.0Hz,1H),1.48-1.40(m,2H).

[0467] Example 59

[0468] Similarly, using Intermediate 5 as the starting material, Example 59 was prepared according to Steps 1-2 of the preparation method of Example 57. LCMS (ESI): (MH) - =397.2; 1H NMR(400MHz,Methanol-d4,ppm)δ7.79(dd,J=8.8Hz,2.0Hz,1H),7.58(dd,J=8.4Hz,3.6Hz,1H),7.36-7.30(m,1 H),4.69-4.64(m,2H),3.32-3.22(m,3H),2.13-2.10(m,2H),1.75(dd,J=18.4Hz,6.4Hz,2H),1.53-1.43(m,2H)

[0469] Example 60

[0470] Similarly, using intermediate 4 as the starting material, Example 60 was prepared according to steps 1-3 of the preparation method of Example 57. LCMS (ESI): [M+H] + =462.0; 1 H NMR (400MHz, DMSO-d6) δ8.19 (s, 1H), 7.82 (s, 2H), 4.74 (d, J = 13.2Hz, 2H), 3.23 (s, 2H), 2.01 (d, J = 11.4Hz, 3H), 1.55 (dd, J = 18.1, 6.2Hz, 2H), 1.33 (q, J = 11.4Hz, 2H).

[0471] Example 61

[0472] Similarly, using intermediate 12 as the starting material, Example 61 was prepared according to steps 1-2 of the preparation method of Example 57. LCMS (ESI): (MH) - =413.2; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.97(d,J=1.6Hz,1H),7.57(d,J=8.8Hz,1H),7.49-7.46(m,1H) ,4.63-4.61(m,2H),3.28-3.22(m,4H),2.18-2.15(m,2H),1.60-1.54(m,2H),1.41-1.38(m,1H).

[0473] Example 62

[0474] Similarly, using intermediate 21 as the starting material, Example 62 was prepared according to steps 1-2 of the preparation method of Example 57. LCMS (ESI): (MH) - =491.2; 1H NMR(400MHz,Methanol-d4,ppm)δ8.20(s,1H),7.68-7.62(m,2H),4.60-4.57(m, 2H),3.24-3.17(m,3H),2.10-2.07(m,2H),1.49-1.44(m,2H),1.35-1.33(m,2H).

[0475] Example 63

[0476] Similarly, using intermediate 5 as the starting material, Example 63 was prepared according to steps 1-2 of the preparation method of Example 57. LCMS (ESI): (MH) - =357.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.77-7.74(m,1H),7.57-7.55(m,1H),7.30-7.29(m,1H),3.71-3.67(m,2H),2.04-2.00(m,2H),1.74-1.69(m,2H).

[0477] Example 64

[0478] Preparation of (3-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid:

[0479] Step 1: Preparation of diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate:

[0480] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (350 mg, 0.59 mmol, 1 eq) was dissolved in N,N-dimethylformamide (10 mL). The reaction system was then cooled to 0°C in an ice-water bath, and sodium hydride (56 mg, 2.35 mmol, 4 eq) was slowly added. After stirring for 10 min, iodomethane (333 mg, 2.35 mmol, 4 eq) was added and the reaction was stirred at room temperature for 4 h. The reaction was monitored for completion by LCMS. The reaction was quenched with ice water (40 mL), the mixture was dried, and extracted three times with dichloromethane (100 mL x 3). The organic phases were combined and concentrated to give the crude title compound (300 mg, 0.39 mmol, 83.7% yield). LCMS (ESI): [M+H] + =609.0,611.0.

[0481] Step 2: Preparation of diethyl (3-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate:

[0482] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate (300 mg, 0.49 mmol, 1 equivalent) was dissolved in trifluoroacetic acid (10 mL). The reaction mixture was then stirred and refluxed at 80°C for 2 hours. The reaction mixture was concentrated under reduced pressure, then ice water (40 mL) was added and extracted three times with dichloromethane (100 mL x 3). The organic phases were combined and concentrated to give the crude title compound (200 mg, yield: 83.0%). LCMS (ESI): [M+H] + =489.1,491.1.

[0483] Step 3: Preparation of (3-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid

[0484] Diethyl (3-((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate (200 mg, 0.41 mmol, 1 equivalent) was dissolved in dichloromethane (20 mL), and trimethylsilyl bromide (5 mL) was added. The reaction was heated to 50°C and stirred for 3 hours. The reaction solution was concentrated, and the crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (82 mg, yield: 46.1%) as a white solid. LCMS (ESI): [MH] - =431.1,433.1; 1 H NMR(400MHz,Methanol-d4,ppm)δ8.24(s,1H),7.78-7.66(m,1H),7.60-7.47( m,1H),4.06-3.90(m,2H),3.96(s,3H),2.11-2.05(m,2H),1.76-1.69(m,2H).

[0485] Example 65

[0486] Preparation of (4-(((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonic acid:

[0487] Step 1: Preparation of di-tert-butyl (4-cyanophenyl)phosphonate:

[0488] 4-Bromobenzonitrile (4 g, 16.48 mmol, 1 eq), di-tert-butyl phosphite (12.8 g, 65.92 mmol, 4 eq), and cesium carbonate (10.74 g, 32.96 mmol, 2 eq) were dissolved in toluene (100 mL). Tetrakistriphenylphosphine palladium (952 mg, 0.82 mmol, 0.05 eq) was added to the reaction mixture under a nitrogen atmosphere. After stirring at 110°C for 16 hours, the solvent was evaporated and purified by flash chromatography (200-300 mesh silica gel, ethyl acetate:petroleum ether = 0-5%) to afford the title compound (1.5 g, 50% purity, 11.5% yield) as a light yellow oil. LCMS (ESI) [M+1-56-56] + =184.1.

[0489] Step 2: Preparation of di-tert-butyl (4-(aminomethyl)phenyl)phosphonate:

[0490] Di-tert-butyl (4-cyanophenyl)phosphonate (1.4 g, 2.37 mol, 50% purity, 1 equivalent) was dissolved in ethanol (20 mL). Under a nitrogen atmosphere, Raney nickel (200 mg) was added to the reaction solution. The hydrogen atmosphere was replaced three times, and the reaction solution was allowed to react at room temperature for 16 hours. The reaction mixture was filtered, and the mother liquor was collected. The solvent was dried and separated and purified by flash chromatography (200-300 mesh silica gel, methanol:dichloromethane = 0%-10%, mixed with 0.1% triethylamine) to obtain the title compound (600 mg, 84.6% yield) as a yellow oily liquid. LCMS (ESI) [M+1-56-56] + =188.1.

[0491] Step 3: Preparation of di-tert-butyl (4-(((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonate:

[0492] Di-tert-butyl (4-(aminomethyl)phenyl)phosphonate (446 mg, 1.5 mmol, 2 equiv) and 2,4-dichloro-7-methoxy-5H-pyrimido[5,4-b]indole (200 mg, 0.75 mmol, 1 equiv) were dissolved in acetonitrile (20 mL). Triethylamine (302 mg, 3.0 mmol, 4 equiv) was added, and the reaction mixture was stirred at 80°C overnight. The solvent was dried by spin-drying, and the mixture was purified by flash chromatography (200-300 mesh silica gel, methanol:dichloromethane = 0-2%) to afford the title compound (80 mg, 20.1% yield) as a dark yellow solid. LCMS (ESI) [M+1] + =531.2.

[0493] Step 4: Preparation of (4-(((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonic acid:

[0494] Di-tert-butyl (4-(((2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonate (70 mg, 0.13 mmol, 1 equivalent) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (1 mL) was added to the reaction mixture under ice-cooling conditions. The reaction mixture was allowed to react overnight at room temperature. LCMS confirmed the reaction was complete. The reaction mixture was filtered, and the filter cake was rinsed with methanol and dried to obtain the desired product (46 mg, 84.5% yield) as a white solid. LCMS (ESI) [M+1] + =419.0. 1 H NMR (400MHz, DMSO-d6) δ10.84(s,1H),7.94(t,J=5.6Hz,1H),7.77(d,J=8.8Hz,1H),7.56(dd,J=12.8,7.6Hz,2H),7 .37(dd,J=8.0,3.2Hz,2H),7.03(d,J=2.4Hz,1H),6.75(dd,J=8.8,2.4Hz,1H),4.67(d,J=5.6Hz,2H),3.75(s,3H).

[0495] Example 66

[0496] Preparation of (4-(((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonic acid:

[0497] Step 1: Preparation of diethyl (4-(((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonate:

[0498] Diethyl (4-(aminomethyl)phenyl)phosphonate (600 mg, 2.47 mmol, 2.6 eq) was added to N-methylpyrrolidone (9 mL), followed by the addition of ethyldi(propan-2-yl)amine (2 mL, 15.5 mmol, 16 eq) and 8-bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole (300 mg, 0.95 mmol, 1 eq). The mixture was heated to 160°C and allowed to react for 2 hours. The reaction system was cooled to room temperature, diluted with ethyl acetate, and extracted three times with ethyl acetate (20 mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative plate (DCM, 100%) to afford the title compound (200 mg, 40% yield) as a yellow solid. LCMS (ESI): [M+H] + =525.1.

[0499] Step 2: Preparation of (4-(((8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonic acid:

[0500] Diethyl (4-((8-bromo-2-chloro-5H-pyrimidin[5,4-b]indol-4-yl)amino)methyl)phenyl)phosphonate (80 mg, 0.15 mmol, 1 equivalent) was added to dichloromethane (6 mL), followed by trimethylsilyl bromide (0.5 mL). The reaction was heated to 50°C for 2 hours. 2 mL of water was added, and the aqueous phase was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia, acetonitrile) to afford the title compound (4.51 mg, 6.31% yield) as a white solid. LCMS (ESI): [MH] - =467.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.15(s,1H),8.15-7.82(m,2H),7.45-7.41(m,1H),7.37-7.35(m,3H),4.79(s,2H).

[0501] Example 67

[0502] Preparation of (3-((2-chloro-8-(dimethylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0503] Step 1: Preparation of diethyl (3-((2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrimido[5,4-B]indol-4-yl)amino)propyl)phosphonate:

[0504] Diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (the intermediate obtained in step 3 of Example 069) (100 mg, 0.17 mmol, 1 equivalent) and 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (51 mg, 0.20 mmol, 1.2 equivalents) and potassium acetate (33 mg, 0.34 mmol, 2 equivalents) were dissolved in 1,4-dioxane (5 mL). After reacting in a microwave at 110° C. under argon protection for 1 hour, saturated aqueous sodium bicarbonate solution (10 mL) was added to the reactant, and the mixture was extracted three times with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 1 / 1)) to give the title compound (90 mg, yield: 82.3%) as a yellow solid. LCMS (ESI): [M+H] + =643.1.

[0505] Step 2: Preparation of diethyl (3-((2-chloro-8-(dimethylamino)-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0506] Diethyl (3-((2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (103 mg, 0.16 mmol, 1 eq) was dissolved in acetonitrile (3 mL). Dimethylamine (0.32 mL, 2 M in tetrahydrofuran, 0.64 mmol, 4 eq) and triethylamine (65 mg, 0.64 mmol, 4 eq) were added. The reaction mixture was heated to 60°C and stirred for 3 hours. Saturated aqueous ammonium chloride (10 mL) was added, and the mixture was extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (silica gel, dichloromethane:ethyl acetate (V / V=10 / 1)) to give the title compound (50 mg, yield: 55.8%) as a colorless oily liquid. LCMS (ESI): [M+H] + =560.2.

[0507] Step 3: (3-((2-Chloro-8-(dimethylamino)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0508] Diethyl (3-((2-chloro-8-(dimethylamino)-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.09 mmol, 1 eq) was dissolved in dichloromethane (3 mL). Trimethylsilyl bromide (136 mg, 0.89 mmol, 10 eq) was then slowly added dropwise to the reaction mixture. After addition, the reaction mixture was heated to 50°C and stirred for 1 hour. The reaction mixture was concentrated, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to yield the title compound (1.3 mg, 3.8% yield) as a white solid. LCMS (ESI): [M+H] + =384.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.52 (d, J = 2.4Hz, 1H), 7.47d, J = 8.8Hz, 1H), 7.24- 7.22(m,1H),3.65-3.61(m,2H),2.66(s,6H),2.10-2.07(m,2H),2.06-2.00(m,2H).

[0509] Example 68

[0510] Preparation of ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0511] Step 1: Preparation of tert-butyl 4-((dimethoxyphosphonyl)methylene)piperidine-1-carboxylate:

[0512] Dissolve tert-butyl 4-oxopiperidine-1-carboxylate (5 g, 25.1 mmol, 1 equiv) in tetrahydrofuran (150 mL). Then, slowly add dimethyl {[bis(methoxymethyl)phosphonyl]methyl}phosphonate (23.3 g, 100.4 mmol, 4 equiv) and sodium hydride (2 g, 50.1 mmol, 4 equiv) in an ice-water bath. Stir the reaction at 0°C for 30 minutes. The reaction mixture is diluted with methanol (100 mL) at 0°C, quenched with ice water, and concentrated. The mixture is extracted three times with ethyl acetate (300 mL x 3). The combined organic phases are washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 5 / 1)) to afford the title compound (6 g, 78.7% yield). LCMS (ESI) [M+H] + =306.3.

[0513] Step 2: Preparation of tert-butyl 4-((dimethoxyphosphonyl)methyl)piperidine-1-carboxylate:

[0514] Dissolve tert-butyl 4-((dimethoxyphosphonyl)methylene)piperidine-1-carboxylate (2 g, 6.55 mmol, 1 equivalent) in methanol (50 mL) and add 10% palladium on carbon (1 g). After replacing the hydrogen atmosphere, the reaction was stirred at room temperature for 16 hours. The reaction mixture was filtered to remove the palladium on carbon, and the filtrate was concentrated under reduced pressure to obtain the crude title compound (1.8 g, 5.86 mmol, 89.5% yield). LCMS (ESI) [M+H] + =308.3.

[0515] Step 3: Preparation of dimethyl (piperidin-4-ylmethyl)phosphonate:

[0516] Dissolve tert-butyl 4-((dimethoxyphosphonyl)methyl)piperidine-1-carboxylate (500 mg, 1.63 mmol, 1 equivalent) in dichloromethane (16 mL), then add trifluoroacetic acid (16 mL). Stir the mixture at room temperature for 1 hour. The reaction is monitored for completion by LCMS. The insoluble solid is filtered off, and the filtrate is concentrated under reduced pressure to yield the crude target compound, dimethyl (piperidin-4-ylmethyl)phosphonate (517 mg, 2.49 mmol). LCMS (ESI) [M+H] + =208.3.

[0517] Step 4: Preparation of 2,4-dichloro-7,9-difluoro-5H-pyrimido[5,4-b]indole:

[0518] 7,9-difluoro-1,5-dihydro-2H-pyrimido[5,4-b]indole-2,4(3H)-dione (150 mg, 0.63 mmol, 1 equivalent) was dissolved in phenylphosphonyl dichloride (8 mL) and heated to 180°C for 2 hours. The reaction solution was diluted with ethyl acetate and adjusted to pH 7 with sodium bicarbonate. The mixture was extracted three times with ethyl acetate (10 mL x 3) and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to afford 2,4-dichloro-7,9-difluoro-5H-pyrimido[5,4-b]indole (173 mg, 100% yield). LCMS (ESI): [MH] - =272.0.

[0519] Step 5: Preparation of dimethyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-B]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0520] Dissolve 2,4-dichloro-7,9-difluoro-5H-pyrimido[5,4-b]indole (98 mg, 0.36 mmol, 1 eq) and dimethyl ((piperidin-4-yl)methyl)phosphonate (74 mg, 0.36 mmol, 1 eq) in acetonitrile (3 mL) and add triethylamine (0.98 mL, 9.74 mmol, 27.1 eq). Heat the reaction to 100°C and continue stirring for 1 hour. Dilute the reaction solution with ethyl acetate (10 mL) and wash with ammonium chloride three times (5 mL x 3). Dry the organic phase over anhydrous sodium sulfate and concentrate in vacuo to obtain the title compound (70 mg, yield: 43.1%). LCMS (ESI): [M+H] + =445.0.

[0521] Step 6: Preparation of ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0522] Dissolve dimethyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (70 mg, 0.16 mmol, 1 equivalent) in dichloromethane (6 mL), then add trimethylsilyl bromide (0.5 mL). Heat the reaction to 50°C and continue stirring for 2 hours. After cooling, the reaction solution was extracted with water (2 mL). The aqueous phase was subjected to reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (6.42 mg, yield: 12.5%). LCMS (ESI): [M+H] + =417.1; 1H NMR(400MHz,Methanol-d4,ppm)δ7.12-7.09(m,1H),6.81-6.76(m,1H),4.79-4.56( m,2H),3.26-3.13(m,2H),2.15-2.08(m,3H),1.66-1.60(m,2H),1.47-1.37(m,2H).

[0523] Example 69

[0524] Similarly, using intermediate 21 as the starting material, Example 69 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =453.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.42 (d, J = 2.8Hz, 1H), 7.65-7.63 (m, 1H), 7.62-7.54 (m, 1H), 3.80-3.77 (m, 2H), 2.06-2.05 (m, 2H), 1.85-1.79 (m, 2H).

[0525] Example 70

[0526] Preparation of (3-((2-chloro-8-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0527] Step 1: Preparation of diethyl (3-((2-chloro-8-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0528] At room temperature, 2,4-dichloro-8-methoxy-5H-pyrimido[5,4-b]indole (90 mg, 0.34 mmol, 1 equivalent) and diethyl (3-aminopropyl)phosphonate (262 mg, 1.34 mmol, 4 equivalents) were dissolved in dioxane (10 mL), and diisopropylethylamine (0.5 mL) was added. The reaction was heated to 120 ° C and stirred for 16 hours. The reaction was monitored for completion by LCMS. The reaction solution was quenched with water (20 mL), and the mixture was extracted with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (90 mg, yield: 61.8%) as a brown oil. LCMS (ESI): [M+H] + =427.0.

[0529] Step 2: Preparation of (3-((2-chloro-8-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0530] At room temperature, diethyl (3-((2-chloro-8-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (90 mg, 0.21 mmol, 1 equivalent) was dissolved in dichloromethane (20 mL). Trimethylsilyl bromide (5 mL) was added and the reaction was heated to 50°C and stirred for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was then purified by reverse-phase HPLC (C18, 10 mmol / L ammonia, acetonitrile) to afford the title compound (15 mg, 19.2% yield) as a white solid. LCMS (ESI): [M+H] + =371.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ7.55 (d, J = 2.4Hz, 1H), 7.43 (d, J = 8.8Hz, 1H), 7.14 -7.11(m,1H),3.87(s,3H),3.70-3.68(m,2H),2.08-2.00(m,2H),1.98-1.76(m,2H).

[0531] Example 71

[0532] Preparation of (3-((2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0533] Step 1: Preparation of diethyl (3-((2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0534] Diethyl (3-((2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (the intermediate obtained in Step 1 of Example 137) (90 mg, 0.14 mmol, 1 equivalent) was dissolved in 1,4-dioxane (10 mL), followed by the addition of trifluoromethyl(1,10-phenanthroline)copper(I) (65.7 mg, 0.21 mmol, 1.5 equivalents) and cesium carbonate (137 mg, 0.42 mmol, 3 equivalents). The mixture was reacted at 50°C under an argon atmosphere for 1 hour. The reaction solution was quenched with saturated chloride solution and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (50 mg, 0.09 mmol, 61.06%). LCMS (ESI) [M+H] + =585.1.

[0535] Step 2: Preparation of diethyl (3-((2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0536] Diethyl (3-((2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (115 mg, 0.2 mmol, 1 equivalent) was dissolved in trifluoroacetic acid (2 mL) and reacted at 100°C for 2 hours. The reaction solution was quenched with saturated sodium bicarbonate and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (80 mg, yield: 87.6%). LCMS (ESI): [M+H] + =465.0.

[0537] Step 3: Preparation of (3-((2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0538] Diethyl (3-((2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (30 mg, 0.06 mmol, 1 equivalent) was dissolved in concentrated hydrochloric acid (3 mL), and the reaction mixture was reacted at 110°C for 2 hours. The reaction solution was purified by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (0.85 mg, yield: 3.22%). LCMS (ESI): [MH] -=407.1; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.45(s,1H),7.74(br,2H),3.72-3.69(m,2H),2.19-2.05(m,2H),2.03-2.01(m,2H).

[0539] Example 72

[0540] Preparation of (3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0541] Step 1: Preparation of diethyl (3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0542] At room temperature, 2,4-dichloro-7,9-difluoro-5H-pyrimido[5,4-b]indole (100 mg, 0.36 mmol, 1 equiv) and diethyl (3-aminopropyl)phosphonate (300 mg, 1.53 mmol, 4.2 equiv) were dissolved in acetonitrile (2 mL), followed by the addition of diisopropylethylamine (141 mg, 1.09 mmol, 3 equiv). The reaction was allowed to proceed at 100°C for 2 hours under nitrogen. Following LCMS monitoring, the reaction solution was quenched with 10 mL of saturated aqueous ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to yield the title compound (110 mg, 69.7% yield). LCMS (ESI): [M+H] + =433.0.

[0543] Step 2: Preparation of (3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonic acid:

[0544] Diethyl (3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (50 mg, 0.12 mmol, 1 equivalent) was dissolved in concentrated hydrochloric acid (3 mL) at room temperature and then heated to 110°C for 2 hours. After completion of the reaction, the reaction was monitored by LCMS and prepared by reverse phase HPLC (C18, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (8.7 mg, yield: 20.0%). LCMS (ESI): [M+H] + =377.1; 1H NMR (400MHz, Methanol-d4) δ7.14 (d, J = 8.4Hz, 1H), 6.78 (t, J = 10.4Hz, 1H), 3.68 (q, J = 6.4Hz, 2H), 2.04-2.00 (m, 2H), 1.76-1.72 (m, 2H).

[0545] Example 73

[0546] Preparation of ((1-(8-bromo-2-chloro-5-methyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0547] Step 1: Preparation of tert-butyl 4-((diethoxyphosphonyl)methylene)piperidine-1-carboxylate:

[0548] Dissolve N-tert-Butyloxycarbonyl-4-piperidone (6.0 g, 30.1 mmol, 1 eq) in tetrahydrofuran (250 mL). Slowly add tetraethyl methylene diphosphonate (17.3 g, 60.2 mmol, 2 eq) and sodium hydride (2.4 g, 60.2 mmol, 2 eq) in an ice-water bath. Stir and react at 0°C for 30 minutes. Quench the reaction with methanol (10 mL) and water (10 mL). The mixture is spin-dried and extracted three times with ethyl acetate (300 mL x 3). The organic phases are combined and concentrated to give the crude title compound (9.3 g, 27.9 mmol, 92.7% yield). LCMS (ESI): [M+Na] + =356.1.

[0549] Step 2: Preparation of tert-butyl 4-((diethoxyphosphonyl)methyl)piperidine-1-carboxylate:

[0550] Dissolve tert-butyl 4-((diethoxyphosphonyl)methylene)piperidine-1-carboxylate (9.3 g, 27.9 mmol, 1 equivalent) in methanol (100 mL). Add 10% (mass percentage) palladium on carbon (4.5 g, 48 wt% (mass percentage)). After replacing the hydrogen atmosphere, stir the reaction at room temperature for 3 hours. The reaction mixture was filtered to remove the palladium on carbon, and the filtrate was concentrated under reduced pressure to obtain the crude title compound (8.1 g, yield: 85.5%). LCMS (ESI): [M+Na] + =358.2.

[0551] Step 3: Preparation of diethyl (piperidin-4-ylmethyl)phosphonate:

[0552] Dissolve tert-butyl 4-((diethoxyphosphonyl)methyl)piperidine-1-carboxylate (3.0 g, 8.95 mmol, 1 equivalent) in dichloromethane (30 mL). Add trifluoroacetic acid (15 mL) and stir at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude title compound (2.1 g, crude yield: 100%) as an oil. LCMS (ESI): [M+H] + =236.1.

[0553] Step 4: Preparation of diethyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0554] 8-Bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole (1.45 g, 4.57 mmol, 1 eq) and diethyl (piperidin-4-ylmethyl)phosphonate (2.15 g, 9.14 mmol, 2 eq) were dissolved in acetonitrile (25 mL). Diisopropylethylamine (1.5 mL) was added and the reaction was heated to 80°C with stirring for 2 hours. The reaction solution was concentrated and then diluted with water (20 mL). The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and dried to give the crude title compound (2 g, yield: 84.9%). LCMS (ESI) [M+H] + =515.1,517.1.

[0555] Step 5: Preparation of diethyl ((1-(8-bromo-2-chloro-5-methyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0556] Diethyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (200.0 mg, 0.39 mmol, 1 eq) was dissolved in 1,4-dioxane (15 mL). Sodium hydride (62 mg, 1.56 mmol, 4 eq) was added under ice-cooling and stirred for 10 minutes. Methyl iodide (221.4 mg, 1.56 mmol, 4 eq) was then added and stirred at room temperature for 3 hours. The reaction was quenched with methanol (10 mL) and water (10 mL), concentrated, and extracted with dichloromethane three times (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (150.0 mg, 0.28 mmol, 72.6% yield). LCMS (ESI) [M+H] + =529.1.

[0557] Step 6: Preparation of ((1-(8-bromo-2-chloro-5-methyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid

[0558] Diethyl ((1-(8-bromo-2-chloro-5-methyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (150 mg, 0.28 mmol, 1 equivalent) was dissolved in dichloromethane (20.0 mL) and trimethylsilyl bromide (5.0 mL) was added. The reaction was heated to 50°C and stirred for 3 hours. The reaction was monitored for completion by LCMS. The filtrate was concentrated and the crude product was slurried with tetrahydrofuran to give the title compound (65 mg, 0.14 mmol, yield: 49.01%) as a white solid. LCMS (ESI): [MH] - =471.1, [M+H] + =473.1; 1 H NMR (400MHz, Methanol-d4) δ8.29(d,J=1.6Hz,1H),7.83(dd,J=9.2,2.0Hz,1H),7.68(d,J=9.2Hz,1H),4.42(d,J=14 .0Hz,2H),3.95(s,3H),3.42(t,J=11.6Hz,2H),2.15-2.13(m,3H),1.77(dd,J=18.4,5.6Hz,2H),1.57-1.54(m,2H).

[0559] Example 74

[0560] Preparation of 1-(2-chloro-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonic acid:

[0561] Step 1: Preparation of diethyl (1-(2-chloro-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate:

[0562] Diethyl (1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate (100 mg, 0.19 mmol, 1 equiv) and potassium vinyl trifluoroborate (26 mg, 0.19 mmol, 1 equiv) were dissolved in 1,4-dioxane (2 mL) and water (0.4 mL). 1,1-Bis(diphenylphosphino)diphenylferric palladium chloride (28 mg, 0.04 mmol, 0.2 equiv) and potassium carbonate (80 mg, 0.58 mmol, 3 equiv) were then added to the mixture. The reaction system was stirred at 100°C under argon for 2 hours. Saturated aqueous ammonium chloride was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by thin-layer chromatography (petroleum ether:ethyl acetate (V / V=3 / 1)) to give the title compound (80 mg, yield: 89.1%) as a white solid. LCMS (ESI): [M+H] + =463.1.

[0563] Step 2: Preparation of 1-(2-chloro-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonic acid:

[0564] Diethyl 1-(2-chloro-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate (30 mg, 0.06 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl bromide (50 mg, 0.32 mmol, 5 equivalents) was then added to the mixture and stirred at 50°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to yield the title compound (7 mg, 28.7% yield) as a white solid. LCMS (ESI): [MH] - =405.2; 1H NMR (400MHz, Methanol-d4) δ8.11(s,1H),7.65(dd,J=8.8,1.8Hz,1H),7.54(d,J=8.8Hz,1H),6.88(dd,J=17.6,11.2Hz,1H),5.76(dd,J=17.6,0.8 Hz,1H),5.16(dd,J=11.0,0.8Hz,1H),4.73(d,J=13.2Hz,2H),3.23(t,J= 12.4Hz,2H),2.20(d,J=13.6Hz,2H),2.15-2.05(m,1H),1.51(dd,J=16.8, 6.5Hz, 2H), 1.38 (ddd, J = 14.8, 13.1, 3.6Hz, 2H).

[0565] Example 75

[0566] Preparation of diphenyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0567] Step 1: Preparation of ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic dichloride:

[0568] At room temperature, ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (100 mg, 0.21 mmol, 1 equivalent) was dissolved in dichloromethane (10 mL). Oxalyl chloride (0.5 mL) was added, followed by 4 drops of N,N-dimethylformamide. The mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction was monitored by TLC and concentrated to give the crude title compound (80 mg, 74.7% yield). LCMS (ESI): [M-2Cl+2MeO+H] + =486.9,488.9.

[0569] Step 2: Preparation of diphenyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0570] Phenol (0.15 mL, 1.61 mmol, 10 equiv) was dissolved in dichloromethane (10 mL) at room temperature. Triethylamine (1 mL) and ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid dichloride (80 mg, 0.162 mmol, 1 equiv) were added. The reaction was stirred at room temperature for 1 hour. LCMS monitored the reaction for completion. The reaction solution was purified by reverse phase HPLC (C18, 10 mmol / L ammonia, acetonitrile) to afford the title compound (40 mg, 40.9% yield) as a white solid. LCMS (ESI): [M+H] + =613.0; 1 H NMR (400MHz, Methanol-d4) δ8.26(d,J=1.7Hz,1H),7.62(dd,J=8.8,1.9Hz,1H),7.51(d,J=8.8Hz,1H) ,7.39-7.36(m,4H),7.29-7.17(m,6H),4.68(d,J=13.6Hz,2H),3.30-3.25(m,2H),2.43-2.31(m,1H), 2.30-2.27(m,1H),2.24(d,J=6.8Hz,1H),2.21-2.13(m,2H),1.60(qd,J=13.3,4.0Hz,2H).

[0571] Example 76

[0572] Similarly, using Intermediate 6 as the starting material, Example 76 was prepared according to Steps 1-2 of the preparation method of Example 57. LCMS (ESI): [M+H] + =441.2; 1 H NMR(400MHz, Methanol-d4)δ7.58(s,1H),7.10(s,1H),4.80-4.69(m,2H),3.95(s,3H),3.92( s,3H),3.23-3.13(m,2H),2.05-2.00(m,3H),1.50(dd,J=16.8,6.8Hz,2H),1.40-1.38(m,2H).

[0573] Example 77

[0574] (2-(5-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)furan-2-yl)ethyl)phosphonic acid:

[0575] Step 1: Preparation of 5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)furan-2-carbaldehyde:

[0576] 2,4-Dichloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indole (330 mg, 0.85 mmol, 1 eq) and (5-formylfuran-2-yl)boronic acid (119 mg, 1.43 mmol, 1.7 eq) were dissolved in a mixture of 1,4-dioxane (10 mL) and water (2 mL). Potassium carbonate (235 mg, 1.7 mmol, 2 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (648 mg, 0.85 mmol, 1 eq) were added to the mixture and reacted at 80°C for 1 hour. The reaction mixture was cooled, saturated aqueous sodium bicarbonate solution (15 mL) was added, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (350 mg, crude yield: 91.8%). LCMS (ESI): [M+H] + =448.0.

[0577] Step 2: Preparation of diethyl (E)-(2-(5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)furan-2-yl)vinyl)phosphonate:

[0578] Tetraethylmethylenebis(phosphonate) (154 mg, 0.54 mmol, 1.5 equiv) and sodium bis(trimethylsilyl)amide (262 mg, 1.43 mmol, 4 equiv) were dissolved in ultra-dry tetrahydrofuran (6 mL) and stirred in an ice-water bath for 0.5 h. A solution of 5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)furan-2-carbaldehyde (160 mg, 0.36 mmol, 1 equiv) in tetrahydrofuran (2 mL) was added to the reaction solution via syringe and stirring continued in an ice-water bath for 1 h. Saturated aqueous sodium chloride solution (10 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield the crude title compound (240 mg) as a brown solid. LCMS: (ESI) [M+H] + =581.6.

[0579] Step 3: Preparation of diethyl (2-(5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)furan-2-yl)ethyl)phosphonate:

[0580] Diethyl (E)-(2-(5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)furan-2-yl)vinyl)phosphonate (240 mg, 0.41 mmol, 1 equiv) was dissolved in anhydrous methanol (5 mL). Tris(triphenylphosphine)rhodium chloride (96 mg, 0.1 mmol, 0.25 equiv) was then added to the mixture. The mixture was purged with a hydrogen balloon and stirred at 25°C for 16 hours. Saturated aqueous ammonium chloride (10 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phase was collected, dried, and concentrated. The crude product was purified by flash chromatography (silica gel, dichloromethane:ethyl acetate (V / V = 1 / 1)) to afford the title compound (120 mg, 0.21 mmol, 49.83%) as a yellow solid. LCMS: (ESI) [M+H] + =583.7.

[0581] Step 4: Preparation of (2-(5-(2-chloro-7-methoxy-5H-pyrimido[5,4-b]indol-4-yl)furan-2-yl)ethyl)phosphonic acid:

[0582] Diethyl (2-(5-(2-chloro-7-methoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)furan-2-yl)ethyl)phosphonate (50 mg, 0.09 mmol, 5 equivalents) was dissolved in acetonitrile (3 mL). Trimethylsilyl bromide (3 mL) was then added to the mixture and stirred at 70°C for 1 hour. The reaction was monitored by LCMS. The reaction solution was directly spin-dried and 2 mL of acetonitrile was added dropwise. A solid precipitated, which was filtered and dissolved in 2 mL of a mixture of tetrahydrofuran and DMF. The solid was then purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to yield the title compound (1.9 mg, 5.4% yield) as a pale yellow solid. LCMS: (ESI) [M+H] + =452.1; 1H NMR(400MHz, Methanol-d4)δ8.10(d,J=8.4Hz,1H),7.41(d,J=1.6Hz,1H),7.32(s,1H),6.93 (d, J=8.0Hz, 1H), 6.43 (d, J=1.2Hz, 1H), 3.96 (s, 3H), 3.19-3.13 (m, 2H), 2.06-1.96 (m, 2H).

[0583] Example 78

[0584] Preparation of ((1-(8-bromo-2-(methyl-d2)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0585] Step 1: Preparation of 8-bromo-2-(methyl-d3)-5H-pyrimido[5,4-b]indole-4-ol:

[0586] Ethyl 3-amino-5-bromo-1H-indole-2-carboxylate (500 mg, 1.77 mmol, 1 equivalent) was dissolved in a solution of hydrogen chloride in 1,4-dioxane (5 mL), followed by the addition of deuterated acetonitrile (1 mL). Under nitrogen, the reaction mixture was reacted at 140°C for 2 hours. The reaction mixture was diluted with ethyl acetate and quenched with saturated ammonium chloride solution. The mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (300 mg, yield: 60.4%) as a yellow oil. LCMS (ESI): [M+H] + =281.0.

[0587] Step 2: Preparation of 8-bromo-4-chloro-2-(methyl-d3)-5H-pyrimido[5,4-b]indole:

[0588] 8-Bromo-2-(methyl-d3)-5H-pyrimido[5,4-b]indole-4-ol (100 mg, 0.36 mmol, 1 equivalent) was dissolved in phthaloyl chloride (2 mL) and reacted at 180°C under nitrogen for 2 hours. The reaction solution was diluted with ethyl acetate and quenched with saturated sodium bicarbonate solution on ice. The solution was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (100 mg, 93.8% yield) as a yellow-brown solid. LCMS (ESI): [MH] - =297.0,299.0.

[0589] Step 3: Preparation of diethyl ((1-(8-bromo-2-(methyl-d3)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0590] 8-Bromo-4-chloro-2-(methyl-d3)-5H-pyrimido[5,4-b]indole (100 mg, 0.33 mmol, 1 eq) and diethyl (piperidin-4-ylmethyl)phosphonate (235 mg, 1 mmol, 3 eq) were dissolved in acetonitrile (2 mL), followed by the addition of diisopropylethylamine (129 mg, 1 mmol, 3 eq). The mixture was reacted at 100°C for 2 hours under nitrogen. The reaction mixture was quenched with saturated ammonium chloride solution and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to afford the title compound (100 mg, yield: 60.8%). LCMS (ESI): [MH] - =496.0,498.0.

[0591] Step 4: Preparation of ((1-(8-bromo-2-(methyl-d2)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0592] Diethyl ((1-(8-bromo-2-(methyl-d3)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (50 mg, 0.1 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL), followed by the addition of trimethylsilyl bromide (76.8 mg, 0.5 mmol, 5 equivalents). The reaction mixture was incubated at 50°C for 2 h. LCMS monitored the reaction completion, followed by quenching with water, concentration in vacuo, and analysis by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to afford the title compound (2.48 mg, 5.6% yield). LCMS (ESI): [MH] - =439.1; 1 H NMR(400MHz,Methanol-d4)δ8.31(s,1H),7.56 -7.49(m,2H),4.70-4.64(m,2H),3.21-3.13(m,2H),2.58-2.55(m,1H),2.21-2 .11(m,2H),2.09-2.01(m,1H),1.50(dd,J=16.8,6.8Hz,2H),1.42-1.33(m,2H).

[0593] Example 79

[0594] ((1-(2-Chloro-8-ethyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid

[0595] Step 1: Preparation of diethyl ((1-(2-chloro-8-ethyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0596] Diethyl (1-(2-chloro-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate (50 mg, 0.11 mmol, 1 eq) was dissolved in methanol (10 mL). Tris(triphenylphosphine)rhodium chloride (20 mg, 0.022 mmol, 0.2 eq) was then added to the mixture. The reaction system was replaced with hydrogen and stirred for 16 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the crude title compound (50 mg, crude yield: 99.6%) as a brown oil. LCMS (ESI): [M+H] + =464.7.

[0597] Step 2: Preparation of ((1-(2-chloro-8-ethyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0598] Diethyl ((1-(2-chloro-8-ethyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (50 mg, 0.11 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl bromide (82 mg, 0.54 mmol, 5 equivalents) was then added to the mixture, and the mixture was stirred at 50°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting brown crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to yield the title compound (16 mg, 36.4% yield) as a white solid. LCMS (ESI): [MH] - =407.2, [M+H] + =409.11; 1H NMR (400MHz, Methanol-d4) δ7.95 (s, 1H), 7.50 (d, J = 8.4Hz, 1H), 7.40 (dd, J = 8.4, 1.6Hz, 1H),4.64(d,J=13.6Hz,2H),3.23(t,J=12.0Hz,2H),2.80(q,J=7.2Hz,2H),2.14-2 .10(m,3H),1.67(dd,J=18.0,6.4Hz,2H),1.48-1.42(m,2H),1.31(t,J=7.2Hz,3H).

[0599] Example 80

[0600] Preparation of phenyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0601] At room temperature, diphenyl ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (40 mg, 0.07 mmol, 1 eq) was dissolved in water (2 mL) and tetrahydrofuran (2 mL). Sodium hydroxide (20 mg, 0.56 mmol, 8 eq) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was monitored for completion by LCMS. The reaction mixture was then purified by reverse phase HPLC (C18, 10 mmol / L ammonia, acetonitrile) to afford the title compound (10 mg, 0.02 mmol, 28.55% yield) as a white solid. LCMS (ESI): [M+H] + =537.1; 1 H NMR (400MHz, Methanol-d4) δ8.26(d,J=1.2Hz,1H),7.61(dd,J=8.8,2.0Hz,1H),7.52(d,J=8.8Hz,1H),7.19-7.20(m,4H),7.03(t, J=7.6Hz,1H),4.63(d,J=13.2Hz,2H),3.25(t,J=11.2Hz,2H),2.25-2.14(m,3H),1.71(dd,J=18.0,6.4Hz,2H),1.47-1.28(m,2H).

[0602] Example 81

[0603] Similarly, using intermediate 14 as the starting material, Example 81 was prepared according to steps 4-6 of the preparation method of Example 68. LCMS (ESI): [M+H] + =417.0;1 H NMR (400MHz, DMSO-d6) δ11.93(s,1H),7.88(dd,J=8.7,4.4Hz,1H),7.34-7.18(m,1H),4.51(d,J =13.2Hz,2H),3.15(t,J=12.5Hz,2H),2.07-1.91(m,3H),1.63-1.51(m,2H),1.44-1.29(m,2H).

[0604] Example 82

[0605] Preparation of ((1-(2-chloro-8-ethoxy-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0606] Step 1: Preparation of diethyl ((1-(2-chloro-5-((4-methoxyphenyl)methyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate:

[0607] Diethyl ((1-(8-bromo-2-chloro-5-((4-methoxyphenyl)methyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)methyl)phosphonate (0.998 g, 1.57 mmol, 1 eq) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,12-dioxoborolane (399 mg, 1.57 mmol, 1 eq) were added. 1,4-dioxane (5 mL) and potassium acetate (154 mg, 1.57 mmol, 1 equiv) were dissolved in 1,4-dioxane (5 mL). [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) chloride (127 mg, 0.157 mmol, 0.1 equiv) was then added. Under argon protection, the reaction solution was heated to 110°C in a microwave oven and stirred for 1 hour. Saturated aqueous ammonium chloride (10 mL) was then added, and the mixture was extracted three times with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 10 / 1)) to afford the title compound (900 mg, 83.8% yield) as a yellow solid. LCMS (ESI): [M+H] + =683.3.

[0608] Step 2: Preparation of diethyl ((1-(2-chloro-8-hydroxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0609] Diethyl ((1-(2-chloro-5-((4-methoxyphenyl)methyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate (100 mg, 0.15 mmol, 1 eq) was dissolved in anhydrous ethanol (3 mL). Lithium hydroxide (10 mg, 0.42 mmol, 2.78 eq) and hydrogen peroxide (0.5 ml, 14.7 mmol, 98 eq) were added and stirred at room temperature for 1 hour. The pH was adjusted to 6-7 with 1 M dilute hydrochloric acid, and the mixture was extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined to give the title compound (100 mg, yield: 35.4%) as a colorless oily liquid. LCMS (ESI): [M+H] + =573.3.

[0610] Step 3: Preparation of diethyl ((1-(2-chloro-8-ethoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0611] Diethyl ((1-(2-chloro-8-hydroxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (130 mg, 0.23 mmol, 1 equiv) was dissolved in anhydrous tetrahydrofuran (5 mL). Sodium hydride (5.44 mg, 0.23 mmol, 1 equiv) was added and stirred at room temperature for 5 minutes. Ethyl iodide (35 mg, 0.23 mmol, 1 equiv) was then added. After stirring at room temperature for 1 hour, saturated aqueous ammonium chloride (10 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 5 / 1)) to afford the title compound (110 mg, 80.7% yield) as a yellow solid. LCMS(ESI):[M+H] + =601.3.

[0612] Step 4: Preparation of ((1-(2-chloro-8-ethoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0613] Diethyl ((1-(2-chloro-8-ethoxy-5-(4-methoxybenzyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (60 mg, 0.1 mmol, 1 equivalent) was dissolved in dichloromethane (3 mL), and trimethylsilyl bromide (0.2 ml, 1.52 mmol, 15 equivalents) was added. The reaction solution was heated to 50°C under argon protection and stirred for 2 hours. LCMS monitored the reaction completion. Water (1 mL) was added to the reaction, and the aqueous phase was prepared by reverse phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (3 mg, yield: 7.1%) as a white solid. LCMS (ESI): [M+H] + =425.2; 1 H NMR (400MHz, Methanol-d4) δ8.51(s,1H),7.59-7.55(m,2H),4.69-4.66(m,2H),4.17(q,J=7.6Hz,2H),3.23-3.15(m,2H),2.23(t,J=7.6Hz,3H),2.02- 1.98(m,3H),1.48-1.44(m,2H),1.38-1.27(m,2H).

[0614] Example 83

[0615] Similarly, using intermediate 22 as the starting material, Example 83 was prepared according to steps 5-6 of the preparation method of Example 68. LCMS (ESI): (MH) - =447.2; 1 H NMR (400MHz, Methanol-d4) δ8.28(s,1H),7.90(d,J=8.8Hz,1H),7.53(d,J=8.8Hz,1H),4.72-4.6 9(m,2H),3.28-3.23(m,2H),2.15-2.13(m,3H),1.69(dd,J=18.0,6.0Hz,2H),1.50-1.43(m,2H).

[0616] Example 84

[0617] Similarly, using Intermediate 22 as the starting material, the intermediate ((1-(2-chloro-8-chloro-5H-pyrimido[5,4-B]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid dimethyl ester was obtained according to Step 5 of the preparation method of Example 68. Methylation and phospholipid hydrolysis reactions were then carried out in sequence according to Steps 5-6 of the preparation method of Example 73 to prepare Example 84. LCMS (ESI): [M+H]+ =463.2; 1 H NMR(400MHz,Methanol-d4)δ8.30(s,1H),7.69-7.60(m,2H),4.08-4.04(m,5H), 3.13(t,J=10.4Hz,2H),2.16-2.10(m,3H),1.71-1.66(m,2H),1.59-1.55(m,2H).

[0618] Example 85

[0619] Preparation of ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0620] Step 1: Preparation of diethyl ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0621] Diethyl (1-(2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)phosphonate (100 mg, 0.15 mmol, 1 equiv) was dissolved in N,N-dimethylformamide (2 mL) at room temperature. Cesium carbonate (95.41 mg, 0.29 mmol, 2 equiv) and trifluoromethyl(1,10-phenanthroline)copper(I) (68.69 mg, 0.22 mmol, 1.5 equiv) were added and reacted at 50°C under argon for 16 h. After completion of the reaction, as monitored by LCMS, the reaction solution was quenched with saturated aqueous ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give diethyl ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (50 mg, 0.08 mmol, 54.64%). LCMS (ESI): [MH] - =625.1.

[0622] Step 2: Preparation of ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0623] Diethyl ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (40 mg, 0.06 mmol, 1 equiv) was dissolved in dichloromethane (1 ml) at room temperature, and trimethylsilyl bromide (97.97 mg, 0.64 mmol, 10 equiv) was added. The mixture was reacted at 50°C for 2 h. After completion of the reaction, as monitored by LCMS, the reaction solution was concentrated under reduced pressure. The crude product was separated and purified by reverse phase preparative method to give ((1-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (1.1 mg, yield: 3.7%). LCMS (ESI): [MH] - =447.2; 1 H NMR(400MHz,Methanol-d4)δ8.48(s,1H),7.78-7.74(m,2H),4.73-4.63(m,2H),3.2 5-3.21(m,2H),2.21-2.15(m,3H),1.61(dd,J=17.6,6.4Hz,2H),1.47-1.39(m,2H).

[0624] Example 86

[0625] Preparation of (3-(2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid:

[0626] Step 1: Preparation of diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate:

[0627] At room temperature, diethyl (3-((8-bromo-2-chloro-5-(4-methoxybenzyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (500 mg, 0.84 mmol, 1 equiv) was dissolved in 1,4-dioxane (10 mL). Diboron (pinacol diboron) (250 mg, 0.98 mmol, 1.17 equiv), [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) chloride (100 mg, 0.14 mmol, 0.16 equiv), and potassium acetate (250 mg, 2.55 mmol, 3.04 equiv) were added. Under argon, the reaction mixture was stirred at 110°C for 1 h. After completion of the reaction, the mixture was quenched with saturated ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title compound (400 mg, yield: 74.1%). LCMS (ESI): [M+H] + =643.2.

[0628] Step 2: Preparation of diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate):

[0629] Diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-5H-pyrimidin[5,4-b]indol-4-yl)amino)propyl)phosphonate (120 mg, 0.19 mmol, 1 equiv) was dissolved in N,N-dimethylformamide (2 mL) at room temperature. Cesium carbonate (122 mg, 0.37 mmol, 2 equiv) and trifluoromethyl(1,10-phenanthroline)copper(I) (88 mg, 0.28 mmol, 1.5 equiv) were added. Under argon, the reaction mixture was stirred at 50°C for 16 h. After completion of the reaction, as monitored by LCMS, the reaction mixture was quenched with saturated ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the title compound (100 mg, yield: 91.6%), LCMS (ESI): [M+H] + =585.2.

[0630] Step 3: Preparation of diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate:

[0631] Diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate (100 mg, 0.17 mmol, 1 equiv) was dissolved in tetrahydrofuran (2 mL) at room temperature, and sodium hydride (33 mg, 1.37 mmol, 8 equiv) was added. Under nitrogen, the reaction solution was stirred at room temperature for 0.5 h, followed by the addition of iodomethane (194 mg, 1.37 mmol, 8 equiv). Stirring was continued for 1 h. After completion of the reaction, monitored by LCMS, the reaction solution was quenched with saturated ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to afford the title compound (90 mg, 88.4% yield). LCMS (ESI): [M+H] + =599.2.

[0632] Step 4: Preparation of (3-(2-chloro-8-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid:

[0633] Diethyl (3-(2-chloro-5-(4-methoxybenzyl)-8-(trifluoromethyl)-5H-pyrimidin[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate (40 mg, 0.07 mmol, 1 equiv) was dissolved in dichloromethane (1 mL) at room temperature. Trimethylsilyl bromide (102 mg, 0.67 mmol, 10 equiv) was added and the mixture was allowed to react at 50°C for 2 h. After completion of the reaction, as monitored by LCMS, the reaction solution was concentrated under reduced pressure. The crude product was purified by reverse-phase HPLC (C18, 10 mmol / L aqueous ammonia, acetonitrile) to afford the title compound (1.67 mg, 5.9% yield) as a white solid. LCMS (ESI): [MH] - =421, [M+H] + =423.05; 1 H NMR(400MHz,MeOD)δ8.46(s,1H),7.90(d,J=8.8Hz,1H),7.72(dd,J=8.8,1.6H z,1H),4.02-3.98(m,2H),3.41(s,3H),2.21-2.08(m,2H),1.76-1.72(m,2H). 19 F NMR (376.5 MHz, MeOD) δ-62.14.

[0634] Example 87

[0635] Preparation of (3-((2,8-dichloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid:

[0636] Step 1: Preparation of benzyl (3-(diethoxyphosphono)propyl)carbamate:

[0637] Diethyl (3-aminopropyl)phosphonate (3 g, 15.384 mmol, 1 equivalent) was dissolved in methanol:water (15:15 mL). The reaction mixture was cooled to 0°C, and sodium bicarbonate (3.2 g, 38.10 mmol, 2.5 equivalents) and benzyl chloroformate (7.9 g, 46.10 mmol, 3 equivalents) were added sequentially. The reaction mixture was stirred in an ice bath for 30 minutes. The reaction mixture was warmed to 25°C and stirred for 12 hours. The reaction mixture was quenched with water (100 mL), and the mixture was extracted three times with EA (200 mL x 3). The organic phases were combined, washed with saturated ammonium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (3 g, 9.11 mmol, 60.53%) as a white oil. LCMS: (ESI) [M+H] + =330.1.

[0638] Step 2: Preparation of benzyl (3-(diethoxyphosphono)propyl)(methyl)carbamate:

[0639] Benzyl (3-(diethoxyphosphono)propyl)carbamate (3 g, 9.11 mmol, 1 eq) was dissolved in THF (50 mL). The reaction mixture was cooled to 0°C and sodium hydride (1.46 g, 3.6 mmol, 4 eq) was added portionwise. The reaction mixture was stirred at 0°C for 0.5 hours. Methyl iodide (2 mL, 3.6 mmol, 4 eq) was then added. After the addition was complete, the reaction mixture was warmed to 25°C and stirred for 16 hours. The reaction mixture was carefully quenched with water (50 mL) in an ice bath. The mixture was extracted five times with ethyl acetate (200 mL x 5). The organic phases were combined, washed with saturated ammonium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound (1.5 g, 4.37 mmol, 50.01% yield). LCMS (ESI) [M+H] + =344.1.

[0640] Step 3: Preparation of diethyl (3-(methylamino)propyl)phosphonate:

[0641] Benzyl (3-(diethoxyphosphono)propyl)(methyl)carbamate (1.5 g, 4.37 mmol, 1 equivalent) was dissolved in methanol (30 mL) and 10% palladium on carbon (700 mg) was added. Under hydrogen, the reaction mixture was heated to 25°C and stirred for 3 hours. The reaction mixture was filtered, and the residue was washed three times with methanol. The combined filtrates were concentrated under reduced pressure to obtain the crude title compound (600 mg, 2.87 mmol, 65.6% yield). LCMS (ESI) [M+H] + =210.1.

[0642] Step 4: Preparation of diethyl (3-((2,8-dichloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate:

[0643] 2,4,8-Trichloro-5H-pyrimido[5,4-b]indole (100 mg, 0.37 mmol, 1 eq) and diethyl (3-(methylamino)propyl)phosphonate (154 mg, 0.73 mmol, 2 eq) were dissolved in acetonitrile (30 mL). Diisopropylethylamine (1.5 mL) was added and the reaction was heated to 90°C with stirring for 2 hours. The reaction solution was concentrated and then diluted with water (20 mL). The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound (100 mg, yield: 61.2%). LCMS (ESI): [M+H] + =445.0.

[0644] Step 5: Preparation of (3-((2,8-dichloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonic acid:

[0645] Diethyl (3-((2,8-dichloro-5H-pyrimido[5,4-b]indol-4-yl)(methyl)amino)propyl)phosphonate (100 mg, 0.22 mmol, 1 equivalent) was dissolved in dichloromethane (100 mL). Trimethylsilyl bromide (25 mL) was added and the reaction was heated to 50°C and stirred for 3 hours. The filtrate was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (17 mg, 19.5% yield) as a white solid. LCMS (ESI): [M+H] + =389.1; 1H NMR (400MHz, Methanol-d4) δ8.07(d,J=2.0Hz,1H),7.73(d,J=8.8Hz,1H),7.43(dd,J= 8.8,2.0Hz,1H),3.99-3.95(m,2H),3.38(s,3H),2.14-2.02(m,2H),1.74-1.67(m,2H).

[0646] Example 88

[0647] Similarly, using intermediate 15 as the starting material, Example 88 was prepared according to steps 1-6 of the preparation method of Example 68. LCMS (ESI): [M+H] + =417.1; 1 H NMR (400MHz, DMSO-d6) δ7.94(dd,J=10.2,8.0Hz,1H),7.55(dd,J=10.9,6.7Hz,1H),4.47(d,J=12.8Hz,2H), 3.15(t,J=12.4Hz,2H),2.15-1.91(m,3H),1.49(dd,J=17.7,6.3Hz,2H),1.32(td,J=13.9,13.3,7.3Hz,2H).

[0648] Example 89

[0649] Similarly, using intermediate 14 as the starting material, Example 89 was prepared according to steps 1-6 of the preparation method of Example 68. LCMS (ESI): [M+H] + =439.1; 1 H NMR(400MHz,DMSO-d6)δ11.77(s,1H),7.95-7.92(m,1H),7.88-7.85(m,1H),7.36( d,J=8.0Hz,2H),7.32-7.22(m,3H),4.71(d,J=5.2Hz,2H),2.97(d,J=21.2Hz,2H).

[0650] Example 90

[0651] Similarly, using intermediate 14 as the starting material, Example 90 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =377.1; 1H NMR(400MHz,DMSO-d6)δ11.74(s,1H), 7.83(dd,J=8.8,4.0Hz,1H),7.61(t,J=5.2Hz,1H),7.32-7.19(m,1H),3.65-3.51(m,2H),1.94-1.79(m,2H),1.76-1.60(m,2H).

[0652] Example 91

[0653] Similarly, starting from Intermediate 15, chlorination was performed according to Step 1 of the preparation method of Example 1, and then Steps 3-4 of the preparation method of Example 65 to prepare Example 91. LCMS (ESI): [M+H] + =439.1; 1 H NMR (400MHz, DMSO-d6) δ11.31(s,1H),8.14(t,J=5.5Hz,1H),8.03(dd,J=10.3,8.1Hz,1H),7.84(dd,J=11.2, 6.7Hz, 1H), 7.34 (d, J = 7.8Hz, 2H), 7.25 (dd, J = 8.2, 2.4Hz, 2H), 4.69 (d, J = 5.3Hz, 2H), 2.96 (d, J = 21.4Hz, 2H).

[0654] Example 92

[0655] Similarly, starting from Intermediate 22, Example 92 was prepared according to Steps 4-5 of the preparation method of Example 87. LCMS (ESI): [M+H] + =423.1; 1 H NMR (400MHz, Methanol-d4) δ8.25(d,J=2.0Hz,1H),7.76(d,J=8.8Hz,1H),7.48(dd,J= 8.8,2.0Hz,1H),4.03-3.99(m,2H),3.45(s,3H),2.17-2.11(m,2H),1.76-1.73(m,2H).

[0656] Example 93

[0657] Similarly, starting from Intermediate 23, chlorination was performed according to Step 1 of the preparation method of Example 1, and then according to Steps 3-4 of the preparation method of Example 65 to prepare Example 93. LCMS (ESI): [M+H] + =411.1;1 H NMR (400MHz, DMSO-d6) δ7.32-7.27(m,1H),7.02-6.97(m,1H),3.60-3.57(m,2H),1.98-1.91(m,2H),1.75-1.63(m,2H).

[0658] Example 94

[0659] Preparation of ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0660] Step 1: Preparation of diethyl ((1-(2-(trifluoromethyl)-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0661] At room temperature, diethyl ((1-(8-bromo-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (200 mg, 0.36 mmol, 1 equiv), potassium vinyl trifluoroborate (146 mg, 1.09 mmol, 3 equiv), 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (80 mg, 0.11 mmol, 0.3 equiv), and potassium carbonate (150 mg, 1.09 mmol, 3 equiv) were dissolved in a mixture of dioxane (5 mL) and water (1 mL). The atmosphere was replaced with argon three times, and the reaction system was heated to 100°C under argon protection and stirred for 4 hours. Completion of the reaction was monitored by LCMS. The reaction solution was cooled to room temperature and quenched with aqueous solution. The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude title compound, diethyl ((1-(2-(trifluoromethyl)-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (150 mg, 0.30 mmol, 83.0%), as an oil. LCMS (ESI): [M+H] + =497.3.

[0662] Step 2: Preparation of diethyl ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0663] At room temperature, diethyl ((1-(2-(trifluoromethyl)-8-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (150 mg, 0.30 mmol, 1 equivalent) was dissolved in methanol (10 mL), and triphenylphosphine rhodium chloride (80 mg) was added. Under a hydrogen atmosphere, the reaction was heated to 25°C and stirred for 2 hours. The reaction was monitored for completion by LCMS. The reaction mixture was filtered, and the residue was washed three times with methanol. The filtrates were combined and concentrated under reduced pressure to give the crude target compound, diethyl ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (150 mg, 99.6% yield) as an oil. LCMS (ESI): [M+H] + =499.1.

[0664] Step 3: Preparation of ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0665] At room temperature, diethyl ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (150 mg, 0.30 mmol, 1 equivalent) was dissolved in dichloromethane (30 mL). Trimethylsilyl bromide (2 mL) was added and the reaction was heated to 50°C and stirred for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was dried and the crude product was purified by reverse phase HPLC (C18, 10 mmol / L ammonia, acetonitrile) to afford the title compound ((1-(8-ethyl-2-(trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (16 mg, 13.0% yield) as a white solid. LCMS (ESI): [MH] - =441.1, [M+H] + =443.1; 1 H NMR (400MHz, Methanol-d4) δ8.12(s,1H),7.55(d,J=8.4Hz,1H),7.44(dd,J=8.8,2.8Hz,1H),4.70(d,J=13.2Hz,2H),3.25-3 .22(m,2H),2.82(q,J=7.6Hz,2H),2.14-2.12(m,3H),1.69(dd,J=18.0,6.0Hz,2H),1.50-1.43(m,2H),1.33(t,J=7.6Hz,3H); 19F NMR (376.5MHz, Methanol-d4) δ-70.34.

[0666] Example 95

[0667] Similarly, starting from Intermediate 23, chlorination was performed according to Step 1 of the preparation method of Example 1, and then according to Steps 3-4 of the preparation method of Example 65 to prepare Example 95. LCMS (ESI): [M+H] + =451.2; 1 H NMR (400MHz, Methanol-d4) δ6.87 (dd, J = 9.2, 1.6Hz, 1H), 6.67-6.62 (m, 1H), 3.96-3. 84(m,2H),3.04-2.97(m,2H),1.94-1.92(m,3H),1.54-1.48(m,2H),1.28-1.24(m,2H)

[0668] Example 96

[0669] Preparation of (1-(8-cyclopropyl-2-trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0670] At room temperature, (1-(8-Bromo-2-trifluoromethyl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (100 mg, 0.20 mmol, 1 eq), potassium cyclopropyltrifluoroborate (142 mg, 1.06 mmol, 5 eq), (SP-4-3)-(dicyclohexyl(2',4',6'-tri(isopropyl)(1,1'-biphenyl)-2-yl)phosphine)(methanesulfonic acid)[2'-(methylamino)[1,1'-biphenyl]-2-yl]palladium (70 mg, 0.08 mmol, 0.4 eq), and cesium carbonate (661 mg, 2.03 mmol, 10 eq) were dissolved in a mixture of dioxane (10 mL) and water (2 mL). The atmosphere was replaced with argon three times, and the reaction system was heated to 100°C under argon protection and stirred for 4 hours. The reaction was monitored by LCMS for completion, and the reaction solution was cooled to room temperature and quenched with aqueous solution. The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (C18, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (3.0 mg, yield: 3.3%) as a white solid. LCMS (ESI): [MH] - =453.3; 1H NMR (400MHz, Methanol-d4) δ8.01(s,1H),7.53(d,J=8.8Hz,1H),7.36(dd,J=8.4,1.6Hz,1H),4.68(d,J=13.2Hz,2H),3.2 9-3.22(m,2H),2.19-2.05(m,4H),1.56(dd,J=17.2,6.4Hz,2H),1.48-1.37(m,2H),1.03-0.98(m,2H),0.78-0.74(m,2H); 19 F NMR (376.5MHz, MeOD) δ-70.36.

[0671] Example 97

[0672] Similarly, starting from Intermediate 24, chlorination was performed according to Step 1 of the preparation method of Example 1, and then Steps 3-4 of the preparation method of Example 65 to prepare Example 97. LCMS (ESI): [M+H] + =383.1; 1 H NMR (400MHz, Methanol-d4) δ8.34(s,1H),7.11(dd,J=9.2,1.6Hz,1H),6.71(t,J=9.2Hz,1H),4.65(d,J=13.2Hz,2 H),3.26-3.13(m,2H),2.20(d,J=11.2Hz,2H),2.16-2.09(m,1H),1.52(dd,J=16.8,6.4Hz,2H),1.48-1.35(m,2H).

[0673] Example 98

[0674] Similarly, starting from Intermediate 23, chlorination was performed according to Step 1 of the preparation method of Example 1, and then according to Steps 3-4 of the preparation method of Example 65 to prepare Example 98. LCMS (ESI): [MH] - =471.2; 1 H NMR (400MHz, Methanol-d4) δ7.31 (d, J = 6.8 Hz, 2H), 6.98-6.93 (m, 3H), 6.48-6.43 (m, 1H), 4.91-4.80 (m, 2H), 2.83 (d, J = 19.4Hz, 2H).

[0675] Example 99

[0676] Similarly, starting from Intermediate 23, chlorination was performed according to Step 1 of the preparation method of Example 1, and then Steps 4-5 of the preparation method of Example 87 were followed to prepare Example 99. LCMS (ESI): [M+H] + =425.1; 1 H NMR (400MHz, Methanol-d4) δ7.35 (dd, J=7.6, 2.0Hz, 1H), 6.82-6.75 (m, 1H), 4.02 -3.98(m,2H),3.41(s,3H),2.14-2.06(m,2H),1.77-1.72(dt,J=17.3,6.9Hz,2H).

[0677] Example 100

[0678] Preparation of ((1-(2-ethyl-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0679] Step 1: Preparation of diethyl ((1-(7,9-difluoro-2-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0680] Diethyl ((1-(2-chloro-7,9-difluoro-5H-pyrimidin[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (100 mg, 0.21 mmol, 1 equiv), potassium vinyl fluoroborate (142 mg, 1.06 mmol, 5 equiv), 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (50 mg, 0.63 mmol, 0.2-0.3 equiv), and potassium carbonate (147 mg, 1.06 mmol, 5 equiv) were dissolved in a mixture of dioxane (5 mL) and water (1 mL) at room temperature. The reaction mixture was heated to 100°C under argon and stirred for 16 hours. After completion of the reaction, as monitored by LCMS, the reaction solution was cooled to room temperature and quenched with aqueous solution. The mixture was extracted three times with dichloromethane (100 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and spin-dried to give the crude title compound (90 mg, 92.3% yield) as an oil. LCMS (ESI): [M+H] + =465.1.

[0681] Step 2: Preparation of diethyl ((1-(2-ethyl-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0682] Diethyl ((1-(7,9-difluoro-2-vinyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (90 mg, 0.19 mmol, 1 equivalent) was dissolved in methanol (20 mL) at room temperature. Triphenylphosphine rhodium chloride (40 mg) was added. The reaction system was replaced with a hydrogen atmosphere, heated to 25°C, and stirred for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was filtered, and the residue was washed three times with methanol. The filtrates were combined, concentrated under reduced pressure, and dried to give the crude title compound (70 mg, yield: 77.4%) as an oil. LCMS (ESI): [M+H] + =467.1.

[0683] Step 3: Preparation of ((1-(2-ethyl-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0684] At room temperature, diethyl ((1-(2-ethyl-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (70 mg, 0.15 mmol, 1 equivalent) was dissolved in dichloromethane (30 mL). Trimethylsilyl bromide (3 mL) was added, and the reaction was heated to 50°C and stirred for 2 hours. After completion of the reaction, monitored by LCMS, the reaction solution was dried, and the crude product was purified by reverse-phase HPLC (C18, 10 mmol / L ammonia, acetonitrile) to give the title compound (16 mg, 26.0% yield) as a white solid. LCMS (ESI): [M+H] + =409.1; 1 H NMR(400MHz, Methanol-d4)δ7.27(dd,J=9.2,2.0Hz,1H),7.04-6.99(m,1H),4.98-4.92(m,2H),3.51-3.34(m,2H) ,3.04(q,J=7.2Hz,2H),2.24-2.15(m,3H),1.75(dd,J=18.4,6.0Hz,2H),1.57-1.47(m,2H),1.43(t,J=7.6Hz,3H). 19 F NMR (376.5MHz, Methanol-d4) δ-114.64,-114.66.

[0685] Example 101

[0686] Similarly, using the intermediate ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid dimethyl ester obtained in step 5 of Example 68 and potassium allyl trifluoroborate as starting materials, Example 101 was prepared according to steps 1-3 of the preparation method of Example 100. LCMS (ESI): [M+H] + =425.2; 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),7.14(d,J=9.6Hz,1H),6.96(t,J=8.4Hz,1H),4.44(d,J=13.2Hz,2H),3.08(t,J=12.4Hz,2 H), 2.74 (t, J = 7.2Hz, 2H), 2.03-1.99 (m, 3H), 1.78 (q, J = 7.6Hz, 2H), 1.53-1.48 (m, 2H), 1.36-1.32 (m, 2H), 0.94 (t, J = 7.2Hz, 3H).

[0687] Example 102

[0688] Preparation of ((3-((8-bromo-2-(trifluoromethyl)-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)bicyclo[1.1.1]pentan-1-yl)methyl)phosphonic acid:

[0689] Step 1: Preparation of tert-butyl (3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)carbamate:

[0690] Under ice-cooling conditions, methyl 3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentane-1-carboxylate (1.0 g, 4.15 mmol, 1 equiv) was added to tetrahydrofuran (15 mL), followed by lithium aluminum hydride (6.22 mL, 1.0 M in THF, 6.22 mmol, 1.5 equiv). The reaction mixture was allowed to react at this temperature for 1 hour. Saturated aqueous sodium chloride solution (50 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 3 / 1)) to afford the title compound (800 mg, 90.5% yield). LCMS (ESI): [M+Na] + =236.2.

[0691] Step 2: Preparation of tert-butyl (3-(iodomethyl)bicyclo[1.1.1]pentan-1-ylcarbamate:

[0692] Tert-butyl (3-(hydroxymethyl)bicyclo[1.1.1]pentan-1-yl)carbamate (800 mg, 3.75 mmol, 1 eq) was dissolved in toluene (10 mL). Imidazole (511 mg, 7.50 mmol, 2 eq), triphenylphosphine (1.48 g, 5.63 mmol, 1.5 eq), and iodine (1.43 g, 5.63 mmol, 1.5 eq) were then added to the mixture and stirred at room temperature for 1 hour. The reaction was monitored by LCMS. The reaction mixture was filtered, saturated aqueous sodium chloride solution (50 mL) was added to the filtrate, and the mixture was extracted three times with ethyl acetate (50 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a brown crude product. The title compound (750 mg, 67.1% yield) was obtained by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 3 / 1)). LCMS (ESI): [M-56+H] + =268.0.

[0693] Step 3: Preparation of tert-butyl (3-((diethoxyphosphono)methyl)bicyclo[1.1.1]pentan-1-ylcarbamate:

[0694] Tert-butyl (3-(iodomethyl)bicyclo[1.1.1]pentan-1-ylcarbamate (750 mg, 1.34 mmol, 1 equivalent) was dissolved in triethyl phosphite (3 mL) and microwave-dried at 180°C for 1 hour. The reaction was monitored by LCMS. Saturated aqueous sodium chloride solution (30 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily liquid, which was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V=1 / 3)) to give the title compound (600 mg, yield: 77.6%). LCMS (ESI): [M+H] + =334.2.

[0695] Step 4: Preparation of diethyl ((3-aminobicyclo[1.1.1]pentan-1-yl)methyl)phosphonate:

[0696] Tert-butyl (3-((diethoxyphosphono)methyl)bicyclo[1.1.1]pentan-1-ylcarbamate (600 mg, 1.80 mmol, 5 equivalents) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (3 mL) was added to the mixture, and the reaction was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to obtain an oily liquid, which was separated and purified by flash chromatography (silica gel, dichloromethane:methanol (V / V=7 / 1)) to give the title compound (190 mg, yield: 45.3%). LCMS (ESI): [M+H]+ =234.2.

[0697] Step 5: Preparation of diethyl ((3-((8-bromo-2-(trifluoromethyl)-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)bicyclo[1.1.1]pentan-1-yl)methyl)phosphonate:

[0698] Diethyl (3-aminobicyclo[1.1.1]pentan-1-yl)methyl)phosphonate (190 mg, 0.815 mmol, 1 eq) and 8-bromo-4-chloro-2-trifluoromethylpyrimidine (285 mg, 0.815 mmol, 1 eq) were dissolved in acetonitrile (3 mL). Diisopropylethylamine (316 mg, 2.42 mmol, 3 eq) was then added to the mixture, and the reaction was stirred at 80°C for 2 hours. Saturated aqueous sodium chloride solution (30 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by reverse-phase HPLC (C18 column, 0.1% trifluoroacetic acid, acetonitrile) (0.1% TFA) to give the title compound (50 mg, 11.2% yield). LCMS (ESI): [M+H] + =547.2.

[0699] Step 6: Preparation of ((3-((8-bromo-2-(trifluoromethyl)-5H-pyrimidinyl[5,4-b]indol-4-yl)amino)bicyclo[1.1.1]pentan-1-yl)methyl)phosphonic acid:

[0700] Ethyl 3-(8-bromo-2-trifluoromethyl-5H-pyrimidinylindol-4-amino)bicyclo[1.1.1]pentane-1-methyl)phosphonate (50 mg, 0.0916 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl bromide (210 mg, 1.37 mmol, 15 equivalents) was then added to the mixture and stirred at 50°C for 1 hour. The reaction was monitored by LCMS, concentrated under reduced pressure, and purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to give the title compound (20.5 mg, 10.7% yield). LCMS (ESI): [MH] - =489.0; 1 H NMR (400MHz, DMSO-d6) δ12.97-11.75 (m, 1H), 8.76 (s, 1H), 8.19 (d, J = 2.0Hz, 1H), 7.70(d,J=8.4Hz,1H), 7.61(d,J=8.4Hz,1H), 2.24(s,6H), 1.89(d,J=18.0Hz,2H).19 F NMR (376.5MHz, DMSO-d6) δ-67.48.

[0701] Example 103

[0702] Preparation of ((1-(7,9-difluoro-2-isopropyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0703] Step 1: Preparation of diethyl ((1-(7,9-difluoro-2-(prop-1-en-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0704] Diethyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (200 mg, 0.424 mmol, 1 equiv), isopropenylboronic acid pinacol ester (142 mg, 0.84 mmol, 2 equiv), (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (100 mg, 0.2 equiv), and cesium carbonate (415 mg, 1.27 mmol, 3 equiv) were dissolved in a mixture of dioxane (10 mL) and water (2 mL). The reaction was heated to 100°C under argon and stirred for 2 hours. The reaction solution was cooled to room temperature and quenched with aqueous solution. The mixture was extracted three times with dichloromethane (300 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase column chromatography (C18 column, 10 mmol / L aqueous acetic acid, acetonitrile) to obtain the title compound (130 mg, yield: 65.7%) as an oil. LCMS (ESI): [MH] - =477.1.

[0705] Step 2: Preparation of diethyl ((1-(7,9-difluoro-2-isopropyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate

[0706] Diethyl ((1-(7,9-difluoro-2-(prop-1-en-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (80 mg, 0.16 mmol, 1 equivalent) was dissolved in methanol (20 mL) and palladium on carbon (40 mg, 50 wt%) was added. The reaction system was replaced with a hydrogen atmosphere, heated to 25°C, and stirred for 2 hours. The reaction mixture was filtered, and the residue was washed three times with methanol. The filtrates were combined and concentrated under reduced pressure to obtain the crude title compound (70 mg, 91.1% yield) as an oil. LCMS (ESI): [M+H] + =481.1.

[0707] Step 3: Preparation of ((1-(7,9-difluoro-2-isopropyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid

[0708] Diethyl ((1-(7,9-difluoro-2-isopropyl-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (70 mg, 0.146 mmol, 1 equivalent) was dissolved in dichloromethane (30 mL). Silane bromide (3 mL) was added and the reaction was heated to 50°C and stirred for 2 hours. The reaction was monitored for completion by LCMS. The reaction solution was then dried and the crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to yield the title compound (25 mg, 40.4% yield) as a white solid. LCMS (ESI): [MH] - =423.2; 1 H NMR (400MHz, DMSO-d6) δ11.52(s,1H),7.14(d,J=8.8Hz,1H),7.02-6.95(m,1H),4.47(d,J=12.4Hz,2H) ,3.11-3.05(m,3H),2.11-1.99(m,2H),1.55-1.49(m,2H),1.36-1.28(m,2H),1.27(s,3H),1.26(s,3H).

[0709] Example 104

[0710] Similarly, using the intermediate ((1-(7,9-difluoro-2-(prop-1-en-2-yl)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid diethyl ester obtained in step 1 of Example 103 as a starting material, phospholipid hydrolysis was performed according to step 3 of the preparation method of Example 103 to prepare Example 104. LCMS (ESI): (MH)- =421.2; 1 H NMR (400MHz, DMSO-d6) δ11.73(s,1H),7.16(d,J=9.2Hz,1H),6.97(t,J=9.2Hz,1H),6.25(s,1H),5.32(s,1H),4.46(d,J =13.2Hz,2H),3.10(t,J=12.0Hz,2H),2.19(s,3H),2.06-1.96(m,3H),1.52(dd,J=18.0,6.4Hz,2H),1.41-1.31(m,2H).

[0711] Example 105

[0712] Preparation of 3,6-dichloro-9H-carbazole-1-aminopropylphosphonic acid

[0713] Step 1: Preparation of 1-nitro-3,6-dichloro-9H-carbazole

[0714] 3,6-Dichlorocarbazole (600 mg, 2.54 mmol, 1 eq) was dissolved in a mixture of 1,4-dioxane (8 mL) and acetic acid (8 mL). Sodium nitrite (350 mg, 5.08 mmol, 2 eq) was slowly added to the mixture. The reaction mixture was stirred at 110°C for 4 hours. After cooling, the reaction mixture was poured into ice water, slowly stirred for 10 minutes, and filtered. The filter cake was rinsed with water, collected, and dried to obtain the title compound (700 mg, 98% yield).

[0715] Step 2: Preparation of 3,6-dichloro-9-(4-methoxybenzyl)-1-nitro-carbazole

[0716] 2-Nitro-3,6-dichloro-9H-carbazole (700 mg, 2.49 mmol, 1 equivalent) was dissolved in N,N-dimethylformamide (15 mL). Potassium carbonate (1.4 g, 9.96 mmol, 4 equivalents) and p-methoxybenzyl chloride (468 mg, 2.99 mmol, 1.2 equivalents) were added. The reaction mixture was allowed to react at room temperature for 16 hours. The reaction solution was extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 100% to 1 / 1)) to obtain the title compound (750 mg, 75% yield) as an orange-yellow solid.

[0717] Step 3: Preparation of 3,6-dichloro-9-(4-methoxybenzyl)-1-amino-carbazole

[0718] 3,6-Dichloro-9-(4-methoxybenzyl)-1-nitro-carbazole (400 mg, 1.0 mmol, 1 equivalent) was dissolved in tetrahydrofuran (5 mL), and zinc powder (326 mg, 5.0 mmol, 5 equivalents) and acetic acid (5 mL) were added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered, poured into ice water, and extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was isolated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 100% to 1 / 1)) to obtain the title compound (195 mg, 53% yield). 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=2.0Hz,1H),7.69(d,J=8.8Hz,1H),7.60(d,J=2.0Hz,1H),7.39(dd,J=8.8,2.4 Hz,1H),7.02(d,J=8.8Hz,2H),6.82(d,J=2.0Hz,1H),6.79-6.74(m,2H),5.74(s,2H),5.33(s,2H),3.64(s,3H).

[0719] Step 4: Preparation of diethyl 3,6-dichloro-9-(4-methoxybenzyl)-carbazole-1-amino)propyl)phosphonate

[0720] 3,6-Dichloro-9-(4-methoxybenzyl)-1-aminocarbazole (180 mg, 0.48 mmol, 1 equiv), cesium carbonate (316 mg, 0.96 mmol, 2 equiv), and potassium iodide (40 mg, 0.24 mmol, 0.5 equiv) were dissolved in N,N-dimethylformamide (5 mL). Diethyl 3-bromopropylphosphonate (502 mg, 1.94 mmol, 4 equiv) was added, and the reaction mixture was reacted at 90°C for 16 hours. The reaction mixture was poured into ice water and extracted three times with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica gel, methanol:dichloromethane (V / V = 0-5%)) to obtain the title compound (120 mg, 45% yield) as a dark brown oil. LCMS (ESI) [M+1] + =549.2.

[0721] Step 5: Preparation of 3,6-dichloro-9H-carbazole-1-aminopropylphosphonic acid

[0722] Diethyl 3,6-dichloro-9-(4-methoxybenzyl)-carbazole-1-amino)propylphosphonate (115 mg, 0.21 mmol, 1.0 equiv) was dissolved in acetonitrile (8 mL), followed by the addition of trimethylsilyl bromide (320 mg, 2.09 mmol, 5 equiv). Under argon, the reaction mixture was heated to 50°C and stirred for 16 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by reverse-phase HPLC (C18, 10 mmol / L aqueous ammonia, acetonitrile) to afford the title compound (6 mg, 7.6% yield) as a white solid. LCMS (ESI) [M+H] + =373.0; 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.14(d,J=2.2Hz,1H),7.53(d,J=8.6Hz,1H),7.45(d,J=1.7Hz,1H),7 .34(dd,J=8.6,2.1Hz,1H),6.47(d,J=1.6Hz,1H).3.65-3.51(m,2H),1.94-1.79(m,2H),1.76-1.60(m,2H).

[0723] Example 106

[0724] Similarly, using Intermediate 19 as the starting material, Example 106 was prepared according to Steps 1-3 of the preparation method of Example 68. LCMS (ESI): [M+H] + =397.2; 1 H NMR(400MHz,Methanol-d4)δ7.23-7.20(m,1H),6.98-6.93(m,1H),4.91-4.84(m,2H),3.43-3 .38(m,2H),2.70(s,3H),2.25-2.17(m,,3H),1.61(dd,J=18.0,6.8Hz,2H),1.48-1.42(m,2H).

[0725] Example 107

[0726] Similarly, using the intermediate obtained in step 4 of Example 68 as a starting material, Example 107 was prepared according to steps 5-6 of the preparation method of Example 68. LCMS (ESI): [M+H] + =391.1; 1H NMR (400MHz, Methanol-d4) δ7.30(dd,J=9.2,2.0Hz,1H),7.02-6.97(m,1H),4.09-4.05(m,2H),3.65(s,3H),2.18-2.09(m,2H),1.92-1.84(m,2H).

[0727] Example 108

[0728] Similarly, starting from Intermediate 24, chlorination was performed according to Step 1 of the preparation method of Example 1, and then Steps 3-4 of the preparation method of Example 65 were followed to prepare Example 108. LCMS (ESI): [M+H] + =357.1; 1 H NMR (400MHz, Methanol-d4) δ8.34 (s, 1H), 7.34 (dd, J = 11.6, 2.0Hz, 1H), 6.77-6. 71(m,1H),4.00-3.96(m,2H),3.37(s,3H),2.21-2.04(m,2H),1.75-1.67(m,2H).

[0729] Example 109

[0730] Similarly, starting from intermediate 24, chlorination was performed according to step 1 of the preparation method of Example 1, and then steps 3-4 of the preparation method of Example 65 to prepare Example 109. LCMS (ESI): [MH] - =341.2; 1 H NMR (400MHz, Methanol-d4) δ 8.33 (s, 1H), 7.17 (d, J = 8.4Hz, 1H), 6.79 (t, J = 10.0Hz, 1H), 3.69 (t, J = 6.4Hz, 2H), 2.06-2.03 (m, 2H), 1.78-1.70 (m, 2H).

[0731] Example 110

[0732] Similarly, using the phosphonate intermediate (diethyl 3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate obtained in step 1 of Example 72 and potassium vinyl fluoroborate as raw materials, Example 110 was prepared according to steps 1-3 of the preparation method of Example 100. LCMS (ESI): [M+H] + =371.1; 1H NMR (400MHz, Methanol-d4) δ7.04(d,J=10.0Hz,1H),6.56(t,J=10.0Hz,1H),3.68(t,J=6.8 Hz, 2H), 2.84 (q, J = 7.6 Hz, 2H), 2.08-2.01 (m, 2H), 1.69-1.62 (m, 2H), 1.34 (t, J = 7.6 Hz, 3H).

[0733] Example 111

[0734] Similarly, using the phosphonate intermediate (diethyl 3-((2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate obtained in step 1 of Example 72 and isopropenylboronic acid pinacol ester as starting materials, Example 111 was prepared according to steps 1-3 of the preparation method of Example 103. LCMS (ESI): [M+H] + =385.2; 1 H NMR (400MHz, Methanol-d4) δ7.11-7.01(m,1H),6.67(d,J=10.0Hz,1H),3.71(t,J=6.0H z,2H),3.30-3.09(m,1H),2.07-2.00(m,2H),1.68-1.63(m,2H),1.33(d,J=6.8Hz,6H).

[0735] Example 112

[0736] Preparation of phenyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0737] Step 1: Preparation of (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphine dichloride:

[0738] Dissolve (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (150 mg, 0.36 mmol, 1 eq) in dichloromethane (5 mL). Under argon, add oxalyl chloride (114 mg, 0.90 mmol, 2.5 eq) dropwise, followed by 4 drops of N,N-dimethylformamide. The reaction mixture is stirred at 20°C for 1 hour and then concentrated under reduced pressure to afford the title compound (150 mg, 91.9% yield) as a yellow oil.

[0739] Step 2: Preparation of diphenyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0740] (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphine dichloride (150 mg, 0.33 mmol, 1 eq) and diisopropylethylamine (212 mg, 1.65 mmol, 5 eq) were dissolved in dichloromethane (5 mL). A solution of phenol (124 mg, 1.32 mmol, 4 eq) in anhydrous tetrahydrofuran (2 mL) was slowly added dropwise. After stirring at room temperature for 0.5 hour, the mixture was concentrated under reduced pressure and purified by flash chromatography (silica gel, dichloromethane:methanol (V / V=10 / 1)) to give the title compound (120 mg, 63.9% yield) as a yellow oil. LCMS (ESI): [M+H] + =569.2.

[0741] Step 3: Preparation of phenyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate

[0742] Diphenyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (148 mg, 0.26 mmol, 1 equivalent) was dissolved in methanol (2 mL) and water (2 mL). Sodium hydroxide (104 mg, 2.6 mmol, 10 equivalents) was added, and the reaction mixture was stirred at room temperature for 30 minutes. Reverse-phase HPLC (C18 column, 10 mmol / aqueous ammonia, acetonitrile) yielded the title compound (17.36 mg, 13.6% yield) as a white solid. LCMS (ESI): [M+H] + =493.2; 1 H NMR (400MHz, Methanol-d4) δ7.29-7.20(m,4H),7.11-7.04(m,2H),6.79(t,J=10.0Hz,1H),4.58(d,J=1 3.2Hz, 2H), 3.23 (t, J = 11.6Hz, 2H), 2.15-2.13 (m, 3H), 1.73 (dd, J = 18.0, 6.8Hz, 2H), 1.47-1.40 (m, 2H).

[0743] Example 113

[0744] Preparation of ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidin-4-methyl)phosphonic acid:

[0745] Step 1: Preparation of tert-butyl 4-(iodomethyl)-4-methylpiperidine-1-carboxylate:

[0746] Tert-butyl 4-(hydroxymethyl)-4-methylpiperidin-1-carboxylate (2.0 g, 8.7 mmol, 1.0 equiv) was added to tetrahydrofuran (5 mL), followed by triphenylphosphine (3.43 g, 13.1 mmol, 1.5 equiv), iodine (3.32 g, 13.1 mmol, 1.5 equiv), and imidazole (594 mg, 8.7 mmol, 1.0 equiv). The reaction mixture was reacted at 70°C for 16 hours. The reaction mixture was quenched by addition of water (20 mL) and extracted three times with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and added to silica gel for spin drying. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 4 / 1)) to afford the title compound (1.8 g, 60.8% yield) as a yellow oil. LCMS (ESI): [M+H-Boc] + =240.0.

[0747] Step 2: Preparation of tert-butyl 4-(diethoxyphosphonyl)methyl)-4-methylpiperidine-1-carboxylate:

[0748] tert-Butyl 4-(iodomethyl)-4-methylpiperidine-1-carboxylate (1.5 g, 4.4 mmol, 1.0 equiv) was dissolved in N,N-dimethylformamide (5 mL). Triethyl phosphite (1.9 g, 13.3 mmol, 3.0 equiv) and potassium tert-butoxide (1.5 g, 13.3 mmol, 3.0 equiv) were then added to the mixture, and the mixture was stirred at 80°C for 16 hours. The reaction mixture was quenched with water (20 mL) and extracted three times with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and added to silica gel for spin drying. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 3 / 1)) to afford the title compound (400 mg, 25.9% yield) as a yellow oil. LCMS (ESI): [M+H-Boc] + =350.3.

[0749] Step 3: Preparation of diethyl (4-methylpiperidin-4-yl)methyl)phosphonate:

[0750] Dissolve tert-butyl 4-(diethoxyphosphonomethyl)-4-methylpiperidine-1-carboxylate (450 mg, 1.29 mmol, 1.0 equiv) in dioxane (2 mL). Add hydrochloric acid in dioxane (2.6 mL, 10.4 mmol, 8 equiv) to the mixture and stir at room temperature for 2 hours. The reaction mixture is evaporated to dryness under reduced pressure to give the crude title compound (240 mg, 74.7% yield) as a yellow oil. LCMS (ESI): [M+H] + =250.2.

[0751] Step 4: Preparation of diethyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidinylmethyl)phosphonate:

[0752] 2,4-Dichloro-7,9-difluoropyrimido[5,4-b]indole (450 mg, 1.8 mmol, 1.0 equiv) was added to acetonitrile (5 mL), followed by diethyl (4-methylpiperidin-4-yl)methyl)phosphonate (280 mg, 1.8 mmol, 1.0 equiv), and finally N,N-diisopropylethylamine (932 mg, 7.2 mmol, 4.0 equiv). The reaction mixture was allowed to react at 80°C for two hours. LCMS analysis indicated that the reaction was complete, quenched with water (10 mL), and extracted three times with ethyl acetate (15 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and added to silica gel for spin drying. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 1 / 1)) to afford the title compound (719 mg, 81.8% yield) as a white solid. LCMS (ESI): [M+H] + =487.2.

[0753] Step 5: Preparation of ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidin-4-methyl)phosphonic acid:

[0754] Diethyl 1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidinylmethyl)phosphonate (150 mg, 0.31 mmol, 1.0 equiv) was dissolved in dichloromethane (2 mL). Trimethylsilyl bromide (1.9 g, 12.3 mmol, 40 equiv) was then added to the mixture, and the mixture was stirred at 50°C for 0.5 h. The reaction was monitored by LCMS. The reaction solution was filtered, quenched with methanol, and concentrated under reduced pressure. The crude product was purified by reverse-phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to afford the title compound (14.9 mg, 11.2% yield) as a white solid. LCMS (ESI): [M+H] + =431.0; 1 H NMR (400MHz, DMSO-d6, ppm) δ11.81(s,1H),7.19(d,J=10.8Hz,1H),7.04(t,J=8.8Hz,1H),3. 82-3.77(m,4H),1.86-1.83(m,2H),1.74(d,J=18.4Hz,2H),1.57-1.54(m,2H),1.22(s,3H). 19 F NMR (376.5MHz, DMSO-d6) δ-108.99,-113.55.

[0755] Example 114

[0756] Similarly, using the intermediate 2,4-dichloro-7,9-difluoropyrimido[5,4-b]indole obtained in step 4 of Example 68 as a starting material, Example 114 was prepared according to steps 4-5 of the preparation method of Example 113. LCMS (ESI): (MH) - =403.2; 1 H NMR (400MHz, Methanol-d4) δ7.62 (d, J = 8.0 Hz, 2H), 7.35 (dd, J = 8.8, 2.0 Hz, 2H), 7.22 (dd, J = 9.6, 2.4 Hz, 1H), 6.87-6.65 (m, 1H), 3.03 (d, J = 20.4 Hz, 2H).

[0757] Example 115

[0758] Similarly, starting from intermediate 24, chlorination was performed according to step 1 of the preparation method of Example 1, and then steps 4-5 of the preparation method of Example 113 were followed to prepare Example 115. LCMS (ESI): [M+H] + =391.1;1 H NMR (400MHz, Methanol-d4) δ8.24 (s, 1H), 7.65 (d, J = 8.0 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 6.98 (d, J = 10.2 Hz, 1H), 6.45-6.38 (m, 1H), 2.89 (d, J = 19.6 Hz, 2H).

[0759] Example 116

[0760] Similarly, the phosphine ester product of Example 107 was used as the starting material and steps 1-3 of the preparation method of Example 103 were followed to prepare Example 116. LCMS (ESI): [M+H] + =385.2; 1 H NMR(400MHz, Methanol-d4)δ7.34(d,J=8.0Hz,1H),6.76(t,J=9.6Hz,1H),4.04(t,J=8.4Hz,2H), 3.44(s,3H),2.92(q,J=7.6Hz,2H),2.15-2.02(m,2H),1.75-1.67(m,2H),1.39(t,J=7.6Hz,3H).

[0761] Example 117

[0762] Similarly, the phosphine ester product of Example 107 was used as the starting material and steps 1-3 of the preparation method of Example 103 were followed to prepare Example 117. LCMS (ESI): [M+H] + =399.2; 1 H NMR (400MHz, Methanol-d4) δ7.35 (dd, J=9.4, 1.9Hz, 1H), 6.90-6.56 (m, 1H), 4.15-3.83 (m, 2H) ,3.47(s,3H),3.31-3.29(m,1H),2.21-2.05(m,2H),1.81-1.66(m,2H),1.38(d,J=6.8Hz,6H).

[0763] Example 118

[0764] Similarly, using intermediate 12 as the starting material, Example 118 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =375.1; 1H NMR (400MHz, DMSO-d6) δ9.04(s,1H),7.94(d,J=2.0Hz,1H),7.58(d,J=8.8Hz,1H),7.42(dd ,J=8.8,2.1Hz,1H),3.61-3.50(m,2H),2.54(s,1H),2.02-1.86(m,2H),1.80-1.63(m,2H).

[0765] Example 119

[0766] Similarly, using intermediate 26 as the starting material, Example 119 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =381.0; 1 H NMR(400MHz,DMSO-d6)δ11.45(s, 1H),8.46(s,1H),8.06(d,J=2.4Hz,1H),7.64(d,J=8.4Hz,1H),7.54(dd,J=9.2,2.0Hz,1H),4.50(d,J=1 3.2Hz, 2H), 3.13 (t, J = 12.4Hz, 2H), 2.03-1.93 (m, 3H), 1.57 (dd, J = 18.0, 6.4Hz, 2H), 1.41-1.33 (m, 2H).

[0767] Example 120

[0768] Similarly, using intermediate 15 as the starting material, Example 120 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =377.0; 1 H NMR (400MHz, DMSO-d6) δ11.27(s,1H),8.02(dd,J=10.3,8.0Hz,1H),7.85(dd,J=11.2,6. 7Hz, 1H), 7.78 (t, J = 5.3Hz, 1H), 3.61-3.53 (m, 2H), 1.90-1.79 (m, 2H), 1.72-1.60 (m, 2H).

[0769] Example 121

[0770] Similarly, using intermediate 15 as the starting material, Example 121 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =391.0;1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),8.02(dd,J=10.2,8.0Hz,1H),7.52(dd,J=10.8,6.7Hz,1H) ,3.85(t,J=7.9Hz,2H),3.33(s,3H),1.93(tq,J=14.7,7.1Hz,2H),1.67(dt,J=17.5,7.1Hz,2H).

[0771] Example 122

[0772] Similarly, using intermediate 25 as the starting material, Example 122 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =357.1; 1 H NMR(400MHz, DMSO-d6)δ8.87(s,1H),8.19(dd,J=10.2,7.9Hz,1H),7.69(dd,J=10.5, 6.7Hz, 1H), 4.03 (t, J = 7.9Hz, 2H), 3.58 (s, 3H), 2.11-1.90 (m, 2H), 1.83-1.62 (m, 2H).

[0773] Example 123

[0774] Similarly, using intermediate 25 as the starting material, Example 123 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =343.1; 1 H NMR (400MHz, DMSO-d6) δ12.16(s,1H),9.27(t,J=5.5Hz,1H),8.92(s,1H),8.14(dd,J=10.2,7.9H z, 1H), 8.03 (dd, J = 10.9, 6.7Hz, 1H), 3.81 (q, J = 6.4Hz, 2H), 2.02-1.86 (m, 2H), 1.81-1.69 (m, 2H).

[0775] Example 124

[0776] (1-(8-Bromo-2-chlorobenzo[4,5]thieno[3,2-b]pyridin-4-yl)piperidin-4-ylmethyl)phosphonic acid:

[0777] Step 1: Preparation of methyl 5-bromo-3-(3-ethoxy-3-oxopropionamido)benzo[b]thiophene-2-carboxylate:

[0778] Methyl 3-amino-5-bromobenzo[b]thiophene-2-carboxylate (4.8 g, 16.77 mmol, 1 equiv) was dissolved in dichloromethane (70 mL), and triethylamine (2.55 g, 25.16 mmol, 1.5 equiv) was added. The temperature was cooled to 0°C, and ethyl 3-chloro-3-oxopropanoate (3.79 g, 25.16 mmol, 1.5 equiv) was added. The reaction mixture was slowly warmed to 20°C and stirred at this temperature for 16 hours. The reaction mixture was added to water and extracted with dichloromethane (50 mL x 3). The organic phase was washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 3 / 1)) to yield the title compound (5 g, 74%). LCMS (ESI) [M+1] + =368.0.

[0779] Step 2: Preparation of ethyl 8-bromo-2,4-dihydroxybenzo[4,5]thieno[3,2-b]pyridine-3-carboxylate:

[0780] Dissolve methyl 5-bromo-3-(3-ethoxy-3-oxopropionamido)benzo[b]thiophene-2-carboxylate (5 g, 12.49 mmol, 1 equivalent) in tetrahydrofuran (100 mL), cool to 0°C, and add potassium tert-butoxide (2.8 g, 24.99 mmol, 2 equivalents) in portions. Stir at room temperature for 1 hour. Quench the reaction mixture with water, adjust the aqueous phase to pH 3 with 1N hydrochloric acid, and filter to yield the title compound (3.9 g, 85%) as a yellow solid. LCMS (ESI) [M+1] + =368.0.

[0781] Step 3: Preparation of 8-bromobenzo[4,5]thieno[3,2-b]pyridine-2,4-diol:

[0782] Ethyl 8-bromo-2,4-dihydroxybenzo[4,5]thieno[3,2-b]pyridine-3-carboxylate (4 g, 10.86 mmol, 1.0 equivalent) was dissolved in 2N aqueous sodium hydroxide solution (80 mL). The reaction mixture was reacted at 100°C for 5 hours. The reaction mixture was poured into water, adjusted to pH 3 with 2N hydrochloric acid, and filtered. The filter cake was collected to obtain the title compound (3 g, 94% yield) as a yellow solid. LCMS (ESI) [M+1] + =296.0.

[0783] Step 4: Preparation of 8-bromo-2,4-dichlorobenzo[4,5]thieno[3,2-b]pyridine:

[0784] 8-Bromobenzo[4,5]thieno[3,2-b]pyridine-2,4-diol (1 g, 3.38 mmol, 1.0 equivalent) was added to phenylphosphonyl dichloride (10 mL), and the reaction mixture was reacted at 180°C for 5 hours. Saturated sodium carbonate solution was added dropwise to the reaction mixture until pH = 8. The mixture was extracted three times with EA (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the title compound (1.1 g, 97% yield) as a yellow solid. LCMS (ESI) [M+1]+ = 333.9.

[0785] Step 5: Preparation of diethyl (1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-b]pyridin-4-yl)piperidin-4-ylmethyl)phosphonate:

[0786] 8-Bromo-2,4-dichlorobenzo[4,5]thieno[3,2-b]pyridine (300 mg, 0.90 mmol, 1.0 equiv) was dissolved in N-methylpyrrolidone (10 mL). Potassium carbonate (498 mg, 3.6 mmol, 4.0 equiv) and diethyl (piperidin-4-ylmethyl)phosphonate hydrochloride (294 mg, 1.08 mmol, 1.2 equiv) were added. The reaction mixture was reacted at 100°C for 16 hours. The reaction mixture was poured into water and extracted three times with EA (20 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by flash chromatography (silica gel, dichloromethane:methanol (v / v = 50 / 1)) to afford the title compound (350 mg, 73% yield) as a yellow solid. LCMS (ESI) [M+1]+ = 533.0.

[0787] Step 6: Preparation of (1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-b]pyridin-4-yl)piperidin-4-ylmethyl)phosphonic acid:

[0788] Diethyl (1-(8-bromo-2-chlorobenzo[4,5]thieno[3,2-b]pyridin-4-yl)piperidin-4-ylmethyl)phosphonate (300 mg, 0.56 mmol, 1.0 equiv) was dissolved in acetonitrile (15 mL), and trimethylsilyl bromide (432 mg, 2.82 mmol, 5.0 equiv) was added. The reaction mixture was reacted at 50°C for 18 hours. The reaction solution was concentrated, and the crude product was slurried with acetonitrile to give the title compound (182 mg, 68% yield) as a yellow solid. LCMS (ESI) [M+1]+ = 475.0; 1H NMR (400MHz, DMSO-d6) δ8.33(s,1H),8.07(d,J=8.6Hz,1H),7.78(d,J=8.5Hz,1H),6.98(s,1H),3.99(d,J=12.9Hz,2H), 3.06(t,J=11.9Hz,2H), 2.02(d,J=13.3Hz,2H), 1.91(d,J=14.4Hz,1H), 1.59(dd,J=18.0,6.5Hz,2H), 1.46-1.32(m,2H).

[0789] Example 125

[0790] Similarly, using intermediate 28 as the starting material, Example 125 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =434.1; 1 H NMR (400MHz, DMSO-d6) δ7.95(d,J=8Hz,1H),7.52(t,J=8Hz,1H),4.56(d,J=13.4Hz,2H),3.31-3.25(m,2H),2.02(d,J =12.3Hz, 3H), 1.60 (dd, J = 6.1, 18.0Hz, 2H), 1.32-1.40 (m, 2H).

[0791] Example 126

[0792] Similarly, using intermediate 28 as the starting material, Example 126 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =408.0; 1 H NMR (400MHz, DMSO-d6) δ7.94-7.91(m,1H),7.48-7.44(m,1H),3.78(d,J=7.7Hz,2H),3.38(s,3H),1.88(d,J=11.3Hz,2H),1.63-1.55(m,2H).

[0793] Example 127

[0794] Similarly, using intermediate 28 as the starting material, Example 127 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =394.0; 1H NMR(400MHz,DMSO-d6)δ8.59(t,J=5.4Hz,1H),8.02(dd,J=2.2,8.6Hz,1H),7. 51-7.44(m,1H),3.53(q,J=6.6Hz,2H),1.91-1.75(m,2H),1.68-1.54(m,2H).

[0795] Example 128

[0796] Similarly, using intermediate 25 as the starting material, Example 128 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =383.1; 1 H NMR (400MHz, DMSO-d6) δ11.48(s,1H),8.45(s,1H),8.04(dd,J=10.2,8.1Hz,1H),7.54(dd,J=10.8,6.7Hz,1H),4.49(d,J =13.0Hz, 2H), 3.13 (t, J = 12.5Hz, 2H), 2.02 (d, J = 13.0Hz, 3H), 1.57 (dd, J = 18.1, 6.2Hz, 2H), 1.42-1.28 (m, 2H).

[0797] Example 129

[0798] Preparation of (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)(methyl)phosphonic acid

[0799] Step 1: Preparation of tert-butyl 4-(ethoxy(methyl)phosphonyl)methyl)piperidine-1-carboxylate:

[0800] Dissolve tert-butyl 4-(iodomethyl)piperidine-1-carboxylate (500 mg, 2.1 mmol, 1.0 equiv) in diethyl methylphosphonate (2 mL) and microwave at 80°C for 1 hour. Completion of the reaction was monitored by LCMS, followed by concentration under reduced pressure. The crude product was purified by flash chromatography (silica gel, dichloromethane:methanol (V / V = 10 / 1)) to afford the title compound (500 mg, 77.1% yield) as an oily liquid. LCMS (ESI): [M+H] + =306.2.

[0801] Step 2: Preparation of ethyl methyl (piperidin-4-ylmethyl)phosphite:

[0802] Dissolve tert-butyl 4-(ethoxy(methyl)phosphonyl)methyl)piperidine-1-carboxylate (500 mg, 1.6 mmol, 1.0 equivalent) in dichloromethane (3 mL). Then, add trifluoroacetic acid (3 mL) to the mixture and stir at room temperature for 2 hours. The reaction is monitored by LCMS. After concentration under reduced pressure, silica gel is added and the mixture is dried by spin drying. The crude product is purified by flash chromatography (silica gel, dichloromethane:methanol (V / V = 7 / 1)) to obtain the title compound (300 mg, yield: 89.3%) as an oily liquid. LCMS (ESI): [M+H] + =206.2.

[0803] Step 3: Preparation of ethyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)(methyl)phosphinate:

[0804] Ethyl methyl(piperidin-4-ylmethyl)phosphite (300 mg, 1.5 mmol, 1.0 equiv) and 2,4-dichloro-7,9-difluoropyrimido[5,4-b]indole (400 mg, 1.5 mmol, 1.0 equiv) were dissolved in acetonitrile (5 mL). N,N-diisopropylethylamine (567 mg, 4.4 mmol, 3.0 equiv) was then added to the mixture, and the reaction was stirred at 80°C for 2 hours. Saturated aqueous sodium chloride solution (30 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by reverse phase chromatography (C18 column, 0.1% TFA, acetonitrile) to obtain the title compound (150 mg, yield: 23.2%) as a white solid. LCMS (ESI): [M+H] + =443.2.

[0805] Step 4: Preparation of (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-methyl)(methyl)phosphonic acid:

[0806] Ethyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)(methyl)phosphinate (150 mg, 0.34 mmol, 1.0 equiv) was dissolved in acetonitrile (3 mL). Trimethylsilyl bromide (779 mg, 5.1 mmol, 15 equiv) was then added to the mixture and stirred at 50°C for 1 hour. The reaction solution was quenched with methanol and concentrated under reduced pressure to obtain a crude brown oil. The crude product was purified by reverse phase HPLC (C18 column, 10 mmol / L aqueous ammonia, acetonitrile) to obtain the title compound (33.26 mg, yield: 23.7%) as a white solid. LCMS (ESI): [M+H] + =415.0; 1 H NMR (400MHz, DMSO-d6) δ7.30(d,J=9.2Hz,1H),6.98(t,J=10.4Hz,1H),4.58(d,J=12.8Hz,2H),3.17(d,J=11.2 Hz, 2H), 2.06 (d, J = 11.2 Hz, 3H), 1.48 ( dd, J = 13.2, 5.6 Hz, 2H), 1.33 ( d, J = 12.0 Hz, 2H), 1.18 ( d, J = 13.3 Hz, 3H).

[0807] Example 130

[0808] Similarly, using intermediate 25 as the starting material, Example 130 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =397.1; 1 H NMR(400MHz,DMSO-d6)δ8.39(s, 1H),7.22(dd,J=9.6,2.0Hz,1H),6.95(t,J=10.4Hz,1H),3.78(s,4H),1. 97-1.79(m,2H),1.67(d,J=18.4Hz,2H),1.58-1.43(m,2H),1.18(s,3H).

[0809] Example 131

[0810] Similarly, using the intermediate (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidinylmethyl)phosphonic acid diethyl ester obtained in step 4 of Example 113 as a starting material, Example 131 was prepared according to steps 1-3 of the preparation method of Example 103. LCMS (ESI): [M+H] + =425.2;1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),7.14(d,J=8.0Hz,1H),6.97(t,J=9.6Hz,1H),3.81-3.79 (m,4H),2.81-2.76(m,2H),1.88-1.69(m,4H),1.58(s,2H),1.29(t,J=7.6Hz,3H),1.21(s,3H).

[0811] Example 132

[0812] Similarly, using intermediate 26 as the starting material, Example 132 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =341.0; 1 H NMR (400MHz, DMSO-d6) δ8.33(s,1H),7.99(d,J=2.0Hz,2H),7.63(d,J=9.2Hz,1H ),7.45(d,J=8.8Hz,1H),3.62-3.59(m,2H),1.91-1.90m,2H),1.78-1.70(m,2H).

[0813] Example 133

[0814] Similarly, using intermediate 26 as the starting material, Example 133 was prepared according to steps 1-3 of the preparation method of Example 1. LCMS (ESI): [M+H] + =355.0; 1 H NMR (400MHz, DMSO-d6) δ8.32(s,1H),7.97(d,J=2.0Hz,1H),7.72(d,J=8.8Hz,1H),7.42(d d,J=9.6,2.0Hz,1H),3.93-3.88(m,2H),3.22(s,3H),2.33-2.32(s,2H),1.45-1.23(m,2H)

[0815] Example 134

[0816] Preparation of (2-(1-(2-chloro-7,9-difluoro-5H-pyrimidinyl[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)boronic acid:

[0817] Step 1: Preparation of 2-(1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)ethan-1-ol:

[0818] 4-Chloro-7,9-difluoro-2-methyl-5H-pyrimido[5,4-b]indole (100 mg, 0.39 mmol, 1 eq) was dissolved in acetonitrile (5 mL), and N,N-diisopropylethylamine (1 mL) and ethylbis(propan-2-yl)amine (252 mg, 1.95 mmol, 5 eq) were added. The reaction was allowed to react at 100°C for 1 h. After completion of the reaction, saturated aqueous ammonium chloride (10 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (110 mg, yield: 76.9%) as a brown solid. LCMS (ESI): [M+H] + =367.0.

[0819] Step 2: Preparation of 1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-(2-iodoethyl)piperidine:

[0820] 2-(1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)ethan-1-ol (100 mg, 0.27 mmol, 1 equivalent) was dissolved in dichloromethane (1 mL), and imidazole (28 mg, 0.41 mmol, 1.5 equivalents) was added. The reaction was allowed to react at room temperature for 1 hour. After completion of the reaction, saturated aqueous ammonium chloride solution (10 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (70 mg, yield: 54.4%) as a white solid. LCMS (ESI): [M+H] + =477.0.

[0821] Step 3: Preparation of 2-chloro-7,9-difluoro-4-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethyl)piperidin-1-yl)-5H-pyrimido[5,4-b]indole:

[0822] 1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-(2-iodoethyl)piperidine (100 mg, 0.27 mmol, 1 eq) was dissolved in N,N-dimethylformamide (1 mL), followed by the addition of triphenylphosphine (5 mg, 0.02 mmol, 0.12 eq), bis-neopentyl glycol diborate (66 mg, 0.29 mmol, 2 eq), and cuprous iodide (3 mg, 0.01 mmol, 0.1 eq). The reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride (10 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the title compound (50 mg, 69.9% yield) as a white solid. LCMS (ESI): [M+H] + =476.9.

[0823] Step 4: Preparation of (2-(1-(2-chloro-7,9-difluoro-5H-pyrimidinyl[5,4-b]indol-4-yl)piperidin-4-yl)ethyl)boronic acid:

[0824] 1-(2-Chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-(2-iodoethyl)piperidine (100 mg, 0.27 mmol, 1 equivalent) was dissolved in ethanol (1 mL) and water (1 mL), followed by the addition of lithium hydroxide (63 mg, 2.63 mmol, 20.88 equivalents). The reaction system was allowed to react at room temperature for 30 minutes. The reaction solution was filtered, concentrated, and purified by reverse-phase HPLC (C18 column, 0.1% trifluoroacetic acid, acetonitrile) to obtain the title compound (2.0 mg, 3.2% yield) as a white solid. LCMS (ESI): [M+H] + =395.1; 1 H NMR (400MHz, Methanol-d4) δ7.11 (dd, J=11.2, 2.0Hz, 1H), 6.80 (td, J=10.4, 2.0Hz, 1H), 4.60 (d, J=13.6 Hz,2H),3.56-3.48(m,2H),3.24-3.17(m,1H),1.93-1.90(m,3H),1.61-1.56(m,2H),1.38-1.29(m,2H).

[0825] Example 135

[0826] Similarly, using the intermediate 4-chloro-7,9-difluoro-5H-pyrimidin[5,4-b]indole as the starting material, Example 135 was prepared according to steps 1-4 of the preparation method of Example 134. LCMS (ESI): [M+H] + =388.9; 1 H NMR (400MHz, Methanol-d4) δ7.63 (d, J = 8.0 Hz, 2H), 7.46-7.38 (m, 2H), 7.11 (dd, J = 9.2, 1.9 Hz, 1H), 6.80 (td, J = 10.4, 2.0 Hz, 1H), 4.81 (s, 2H).

[0827] Example 136

[0828] Similarly, using the intermediate (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)-4-methylpiperidinylmethyl)phosphonic acid diethyl ester obtained in step 4 of Example 113 as a starting material, Example 136 was prepared according to steps 1-3 of the preparation method of Example 103. LCMS (ESI): [M+H] + =439.2; 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),7.14(dd,J=9.2,2.0Hz,1H),6.95(t,J=10.4Hz,1H),3.86-3.72(m,4H),3.1 3-3.00(m,1H),1.88-1.78(m,2H),1.75(d,J=18.8Hz,2H),1.64-1.50(m,2H),1.29(d,J=6.8Hz,6H),1.22(s,3H).

[0829] Example 137

[0830] Similarly, the phosphonate intermediate (diethyl 3-((2-chloro-8-methoxy-5H-pyrimido[5,4-b]indol-4-yl)amino)propyl)phosphonate obtained in Step 1 of Example 70 was used as the starting material, and Example 137 was prepared according to Steps 1-3 of the preparation method of Example 132. LCMS (ESI): [M+H]+=385.0; 1H NMR (400 MHz, Methanol-d4) δ 7.83-7.81 (m, 1H), 7.55-7.54 (m, 1H), 7.11-7.08 (m, 1H), 4.05-4.01 (m, 2H), 3.87 (s, 3H), 3.39 (s, 3H), 2.13-2.09 (m, 2H), 1.65-1.59 (m, 2H).

[0831] Example 138

[0832] Preparation of (4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenethyl)phosphonic acid:

[0833] Step 1: Preparation of 2-(4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenyl)ethan-1-ol:

[0834] 4-Chloro-7,9-difluoro-5H-pyrimidin[5,4-b]indole (200 mg, 0.83 mmol, 1 equivalent) and 4-(hydroxyethyl)phenylboronic acid pinacol ester (249 mg, 1.0 mmol, 1.2 equivalents) were dissolved in dioxane (2 mL) and water (0.4 mL). 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (61 mg, 0.08 mmol, 0.1 equivalent) and potassium carbonate (346 mg, 2.50 mmol, 3 equivalents) were added to the mixture and reacted at 100 ° C for 2 hours. The reaction mixture was diluted with saturated aqueous ammonium chloride (30 mL), and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by thin-layer preparative plate chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 2 / 1)) to obtain the title compound (200 mg, yield: 74.1%) as a yellow solid. LCMS (ESI): [M+H] + =326.0.

[0835] Step 2: Preparation of 7,9-difluoro-4-(4-(2-iodoethyl)phenyl)-5H-pyrimido[5,4-b]indole:

[0836] 2-(4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenyl)ethan-1-ol (200 mg, 0.61 mmol, 1 equiv) was dissolved in dichloromethane (2 mL). Iodine (83 mg, 1.22 mmol, 2 equiv), triphenylphosphine (320 mg, 1.22 mmol, 2 equiv), and imidazole (310 mg, 1.22 mmol, 2 equiv) were then added to the mixture. The mixture was purged with nitrogen three times and reacted at 50°C for 4 hours. The reaction solution was diluted with saturated aqueous sodium bicarbonate (30 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was isolated and purified by preparative thin-layer chromatography (silica gel, petroleum ether:ethyl acetate (v / v = 2 / 1)) to afford the title compound (110 mg, 41.1% yield) as a yellow solid. LCMS(ESI):[M+H] + =436.0.

[0837] Step 3: Preparation of diethyl (4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenethyl)phosphonate:

[0838] 7,9-Difluoro-4-(4-(2-iodoethyl)phenyl)-5H-pyrimido[5,4-b]indole (90 mg, 0.21 mmol, 1 eq) was dissolved in dioxane (1 mL). Triethyl phosphite (172 mg, 1.03 mmol, 5.0 eq) was then added to the mixture. The mixture was reacted at 130°C under microwave conditions for 2 hours. The reaction mixture was concentrated to afford the crude title compound (80 mg, 86.9% yield) as a yellow liquid. LCMS (ESI): [M+H] + =446.1.

[0839] Step 4: Preparation of (4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenethyl)phosphonic acid:

[0840] Diethyl (4-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)phenethyl)phosphonate (70 mg, 0.16 mmol, 1 equivalent) was dissolved in acetonitrile (1 mL). Trimethylsilyl bromide (241 mg, 1.57 mmol, 10 equivalents) was then added to the mixture. The reaction mixture was heated to 50°C and stirred for 2 hours. After completion of the reaction, monitored by LCMS, water (1 mL) was added to the reaction mixture, and the aqueous phase was separated and purified using reverse-phase preparative HPLC (C18 column, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (32 mg, 51.8% yield) as a white solid. LCMS (ESI): [M+H] + =390.0; 1 H NMR (400MHz, Methanol-d4) δ8.82(s,1H),8.21(d,J=8.0Hz,2H),7.51(d,J=8.4Hz,2H),7.15 (dd,J=11.2,2.0Hz,1H),6.64(td,J=10.0,2.0Hz,1H),3.10-3.04(m,2H),1.90-1.82(m,2H).

[0841] Example 139

[0842] Similarly, using intermediate 23 as the starting material, chlorination reaction was carried out according to step 1 of the preparation method of Example 1, and then steps 1-4 of the preparation method of Example 138 were followed to prepare Example 139. LCMS (ESI): (MH) - =456.2; 1 H NMR (400MHz, Methanol-d4) δ8.15 (s, 1H), 7.96 (d, J = 7.6Hz, 1H), 7.58-7.49 (m, 2H), 7.3 4(d,J=7.4Hz,1H),6.96(td,J=10.0,2.4Hz,1H),3.20-3.07(m,2H),2.07-1.93(m,2H).

[0843] Example 140

[0844] Similarly, starting from Intermediate 25, chlorination reaction was carried out according to Step 1 of the preparation method of Example 1, and then according to Steps 1-4 of the preparation method of Example 138 to prepare Example 140. LCMS (ESI): [M+H] + =390.0; 1H NMR (400MHz, Methanol-d4) δ9.04(s,1H),8.09(s,1H),7.87(d,J=7.8Hz,1H),7.54(d,J=7.2Hz,1H ),7.50-7.47(m,2H),7.33-7.29(m,1H),6.94-6.89(m,1H),3.13-3.05(m,2H),2.03-1.95(m,2H).

[0845] Example 141

[0846] Preparation of diisopropyl 2,2'-((((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphino)di(azanediyl))(2S,2'S)-dipropionate:

[0847] Step 1: Preparation of P-((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-N-((S)-1-isopropoxy-1-oxopropan-2-yl)phosphonamidic acid:

[0848] ((1-(8-Bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid dichloride (100 mg, 0.22 mmol, 1 equiv) was dissolved in dichloromethane (5 mL), and L-alanine isopropyl ester (116 mg, 0.88 mmol, 4 equiv) and diisopropylethylamine (1 mL, 5.74 mmol, 26 equiv) were added. The mixture was stirred under argon for 10 minutes. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase chromatography (C18 column, 10 mmol / aqueous ammonia, acetonitrile) to afford the title compound (80 mg, 68.5% yield) as a white solid. LCMS (ESI): [MH] - =528.3.

[0849] Step 2: Preparation of diisopropyl 2,2'-((((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphono)di(azanediyl))(2S,2'S)-dipropionate:

[0850] P-((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-N-((S)-1-isopropoxy-1-oxopropan-2-yl)phosphamidic acid (200 mg, 0.38 mmol, 1 equiv) was dissolved in dichloromethane (5 mL), and L-alanine isopropyl ester (99 mg, 0.75 mmol, 2 equiv), 1,8-diazabicyclo[5.4.0]undec-7-ene (287 mg, 1.89 mmol, 5 equiv) and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (294 mg, 0.57 mmol, 1.5 equiv) were added. The reaction solution was stirred at 50°C under argon protection for 0.5 hour. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse phase separation (C18 column, 10 mmol / trifluoroacetic acid, acetonitrile) to obtain the title compound (33 mg, yield: 13.6%) as a white solid. LCMS (ESI): [M+H] + =643.2; 1 H NMR(400MHz,Methanol-d4)δ7.11(d,J=8.8Hz,1H),6.81(t,J=10.4Hz,1H),5.04-4 .97(m,2H),4.61-4.57(m,2H),3.98-3.87(m,2H),3.29-3.20(m,2H),2.13-2.07(m, 3H),1.82-1.72(m,2H),1.52-1.43(m,2H),1.39-1.35(m,6H),1.30-1.24(m,12H). 19 F NMR (376.5MHz, MeOD) δ-110.552,-114.254.

[0851] Example 142

[0852] Preparation of ((((1-(1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonyl)bis(oxy)bis(methylene)diisopropylbis(carbonate):

[0853] (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (100 mg, 0.24 mmol, 1 equivalent) was dissolved in acetonitrile (10 mL), and iodomethyl isopropyl carbonate (0.6 mL, 2.4 mmol, 10 equivalents) and potassium carbonate (330 mg, 2.4 mmol, 10 equivalents) were added. The reaction solution was heated to 60°C and stirred for 16 hours. The reaction was monitored for completion by LCMS. The reaction solution was subjected to reverse phase separation (C18 column, acetonitrile) to obtain the title compound (20 mg, yield: 12.8%) as a white solid. LCMS (ESI): [M+H] + =649.1; 1 H NMR (400MHz, Methanol-d4) δ7.06 (dd, J=6.8 1.2Hz,1H),6.79-6.73(m,1H),5.73-5.64(m,4H),4.94-4.88(m,2H),4.58-4.55(m,2H),3.22(t, J=12.0Hz,2H),2.07-2.02(m,3H),1.98(d,J=6.8Hz,2H),1.49-1.47(m,2H),1.31-1.29(m,12H).

[0854] Example 143

[0855] Preparation of (((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)(phenoxy)phosphono)-L-alanine isopropyl ester:

[0856] Step 1: Preparation of phenyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)chlorophosphonate:

[0857] Phenyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (200 mg, 0.41 mmol, 1 eq) was dissolved in dichloromethane (5 mL). Oxalyl chloride (80 mg, 0.81 mmol, 2 eq) was added dropwise under argon, followed by N,N-dimethylformamide (9 mg, 0.12 mmol, 0.3 eq). The reaction mixture was stirred at 20°C for 1 hour and then concentrated under reduced pressure to obtain the title compound (200 mg, 96.4% yield) as a yellow oil. LCMS (ESI): [M-2Cl+2MeO+H] + =507.0.

[0858] Step 2: Preparation of (((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)(phenoxy)phosphono)-L-alanine isopropyl ester:

[0859] Phenyl ((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)chlorophosphonate (200 mg, 0.39 mmol, 1 equiv) was dissolved in dichloromethane (5 mL), followed by the addition of L-alanine isopropyl ester (105 mg, 0.78 mmol, 2 equiv) and diisopropylethylamine (277 mg, 2.15 mmol, 5 equiv). The mixture was stirred at room temperature under argon for 30 minutes. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase HPLC (C18 column, 10 mmol / L trifluoroacetic acid, acetonitrile) to afford the title compound (41.97 mg, 16.1% yield) as a white solid. LCMS(ESI):[M+H] + =606.1; 1 H NMR (400MHz, DMSO-d6) δ11.80 (s, 1H), 7.35 (dd, J = 15.8, 7.4Hz, 2H), 7.28-7.12 ( m,4H),7.12-7.02(m,1H),5.65-5.42(m,1H),4.84(tt,J=12.6,6.3Hz,1H),4.48 (dd,J=19.0,9.6Hz,2H),3.93-3.80(m,1H),3.22(t,J=12.9Hz,2H),2.25-1.98( m,3H),1.88(ddd,J=11.1,9.5,4.6Hz,2H),1.53-1.32(m,2H),1.22-1.02(m,9H).

[0860] Example 144

[0861] Preparation of 2-((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-4-(3-chlorophenyl)-1,3,2-dioxaphosphane 2-oxide:

[0862] Step 1: Preparation of 1-(3-chlorophenyl)propane-1,3-diol:

[0863] Dissolve methyl 3-(3-chlorophenyl)-3-oxopropanoate (1 g, 4.7 mmol, 1 equivalent) in anhydrous methanol (10 mL) and add sodium borohydride (889 mg, 23.5 mmol, 5 equivalents). Heat to 80°C and stir for 1 hour, then concentrate under reduced pressure. The crude product is purified by reverse phase column chromatography (C18 column, water, acetonitrile) to obtain the title compound (0.70 g, yield: 79.8%) as a yellow oily liquid. 1 H NMR (400MHz, DMSO-d6) δ7.36-7.32(m,2H),7.28-7.25(m,2H),5.27(d,J=4.4Hz,1H),4. 69-4.65(m,1H),4.44(s,1H),3.54-3.49(m,1H),3.46-3.38(m,1H),1.79-1.64(m,2H).

[0864] Step 2: Preparation of 2-((1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-4-(3-chlorophenyl)-1,3,2-dioxaphosphane 2-oxide:

[0865] 1-(3-Chlorophenyl)propane-1,3-diol (165 mg, 0.88 mmol, 2 equiv) was dissolved in dichloromethane (5 mL), followed by the addition of diisopropylethylamine (285 mg, 2.2 mmol, 5 equiv) and (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphine dichloride (200 mg, 0.44 mmol, 1 equiv). The mixture was stirred at room temperature under argon for 1 hour. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase HPLC (C18 column, 10 mmol / L trifluoroacetic acid, acetonitrile) to afford the title compound (20 mg, 8.0% yield) as a white solid. LCMS(ESI):[M+H] + =567.0; 1H NMR (400MHz, Methanol-d4) δ7.46 (s, 1H), 7.42-7.31 (m, 3H), 7.11 (d, J = 11.2, 2.4Hz, 1H), 6.81 (td, J =10.0,2.0Hz,1H),5.68(d,J=11.6Hz,1H),4.74-4.68(m,1H),4.61-4.59(m,2H),4.44-4.33(m,1H), 3.26-3.24(m,2H),2.33-2.19(m,3H),2.13-1.99(m,5H),1.60-1.51(m,2H).

[0866] Example 145

[0867] Preparation of diisopropyl 2,2'-((((1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonyl)bis(nitrogendiyl))(2S,2'S)-dipropionate:

[0868] Step 1: Preparation of (1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphine dichloride:

[0869] Dissolve (1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (199 mg, 0.52 mmol, 1 equivalent) in dichloromethane (5 mL) at room temperature. Oxalyl chloride (164 mg, 1.30 mmol, 2.5 equivalents) was added dropwise under argon. Two drops of N,N-dimethylformamide were added. After stirring at room temperature for 0.5 hours, the reaction was monitored for completion by LCMS (methanol). The mixture was concentrated under reduced pressure to yield the title compound (200 mg, 0.48 mmol, 91.75%) as a white solid. LCMS (ESI): [M-2Cl+2MeO+H] + =411.2.

[0870] Step 2: Preparation of P-((1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-N-((S)-1-isopropoxy-1-oxopropan-2-yl)phosphonamidic acid

[0871] (1-(7,9-Difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphine dichloride (150 mg, 0.36 mmol, 1 equiv) and diisopropylethylamine (1 mL, 5.74 mmol, 16 equiv) were dissolved in dichloromethane (5 mL) at room temperature. L-alanine ethyl ester (142 mg, 1.08 mmol, 3 equiv) was added, and the reaction mixture was stirred at room temperature for 30 minutes under argon. LCMS monitored the reaction for completion. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase HPLC (C18, 10 mmol / aqueous ammonia, acetonitrile) to afford the title compound (100 mg, 56.1% yield) as a white solid. LCMS(ESI):[MH] - =494.3.

[0872] Step 3: Preparation of diisopropyl 2,2'-((((1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphono)bis(nitrogendiyl))(2S,2'S)-dipropionate:

[0873] P-((1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)-N-((S)-1-isopropoxy-1-oxopropan-2-yl)phosphamidic acid (200 mg, 0.4 mmol, 1 equiv) was dissolved in dichloromethane (5 mL) at room temperature. L-alanine isopropyl ester (106 mg, 0.81 mmol, 2 equiv), 1,8-diazabicyclo[5.4.0]undec-7-ene (182 mg, 1.2 mmol, 3 equiv) and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (421 mg, 0.81 mmol, 2 equiv) were added. The mixture was stirred at 50°C under argon for 1 h. The reaction was monitored to completion by LCMS. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase HPLC (C18, 10 mmol / trifluoroacetic acid, acetonitrile) to yield the title compound, 2,2'-((((1-(7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonyl)bis(nitrogendiyl))(2S,2'S)-diisopropyl dipropionate (30 mg, yield: 12.2%), as a white solid. LCMS (ESI): [MH] - =607.5, [M+H]+ =609.5; 1 H NMR(400MHz,CD3OD)δ8.56(s,1H),7.26(dd,J=8.8,2.0Hz,1H),7.05-7.00(m,1H),5.00-4.98(m,2H),4.84-4.79(m,2H),3.97-3 .89(m,2H),3.48-3.40(m,2H),2.24-2.22(m,3H),1.81-1.76(m,2H),1.51-1.50(m,2H),1.40-1.36(m,6H),1.33-1.24(m,12H); 19 F NMR(376.5MHz,MeOD)δ-76.996,-107.110,-115.113.

[0874] Example 146

[0875] Preparation of ((((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphino)bis(oxo))bis(methylene)bis(2,2-dimethylpropionate):

[0876] ((1-(8-Bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (product of Example 40) (90 mg, 0.19 mmol, 1 equivalent) was dissolved in acetonitrile (10 mL) at room temperature. Iodomethyl pivalate (1 mL) and diisopropylethylamine (1 mL) were added. The reaction solution was heated to 60°C and stirred for 16 hours. The reaction was monitored for completion by LCMS. The reaction solution was purified by reverse phase HPLC (C18, acetonitrile) to give the title compound (43 mg, yield: 69%) as a white solid. LCMS (ESI): [M+H] + =687.0,689.0; 1 H NMR (400MHz, Methanol-d4, ppm) δ8.23(d,J=12.0Hz,1H),7.62(s,1H),7.53(d,J=8.8Hz,1H),5.72-5.67 (m,4H),4.66(d,J=13.6Hz,2H),3.26-3.24(m,2H),2.08-1.94(m,5H),1.50-1.47(m,2H),1.23(s,18H).

[0877] Example 147

[0878] Preparation of diethyl 2,2'-((((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphino)di(azanediyl))(2S,2'S)-dipropionate:

[0879] Step 1: Preparation of ((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic dichloride:

[0880] ((1-(8-Bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (product of Example 40) (202.3 mg, 0.44 mmol, 1 equivalent) was dissolved in dichloromethane (5 mL) at room temperature. Oxalyl chloride (112 mg, 0.88 mmol, 2 equivalents) was added dropwise under argon, followed by 2 drops of N,N-dimethylformamide as a catalyst. After stirring at room temperature for 1 hour, the reaction was monitored for completion by LCMS (using methanol as solvent). The mixture was concentrated under reduced pressure to obtain the title compound (200 mg, 91.5% yield) as a white solid. LCMS (ESI): [M-2HCl+2MeOH+H] + =487.0,489.0.

[0881] Step 2: Preparation of diethyl 2,2'-((((1-(8-bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphino)di(azanediyl))(2S,2'S)-dipropionate

[0882] ((1-(8-Bromo-2-chloro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid dichloride (99.3 mg, 0.2 mmol, 1 equiv) was dissolved in dichloromethane (5 mL) at room temperature. L-alanine ethyl ester (117 mg, 1.0 mmol, 5 equiv) and diisopropylethylamine (129 mg, 1 mmol, 5 equiv) were added. The reaction was stirred at room temperature under argon for 10 minutes. LCMS monitored the reaction completion. Saturated aqueous ammonium chloride (10 mL) was added to the reaction, and the mixture was extracted three times with dichloromethane (20 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase HPLC (C18, 10 mmol / formic acid, acetonitrile) to afford the title compound (11 mg, 8.4% yield) as a white solid. LCMS(ESI):[M+H] +=657.2;1H NMR (400MHz, Methanol-d4) δ8.27(d,J=1.6Hz,1H),7.62(dd,J=8.8,1.6Hz,1H),7.52(d,J=8.8Hz,1H),4.65(d,J=12.0Hz,2H),4.26-4.09(m,4H) ,4.00-3.90(m,2H),3.26-3.20(m,2H),2.66(dd,J=12.0,9.6Hz,1H),2.2 5-1.98(m,4H),1.81-1.68(m,2H),1.41-1.36(m,6H),1.30-1.25(m,6H).

[0883] Example 148

[0884] Preparation of diethyl 2,2'-((((1-(8-bromo-2-chlorobenzofurano[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphono)bis(azadiyl))(2S,2'S)-dipropionate:

[0885] ((1-(8-Bromo-2-chlorobenzofuran[3,2-d]pyrimidin-4-yl)piperidin-4-yl)methyl)phosphonic acid (60 mg, 0.13 mmol, 1.0 equiv) was dissolved in dry DCM (2 mL). A catalytic amount of DMF was added, the temperature was cooled to 0°C, and oxalyl chloride (165 mg, 1.30 mmol, 10 equiv) was added dropwise. The reaction was allowed to react at room temperature for 2 h. The reaction solution was then directly spin-dried to obtain the crude acid chloride. L-Alanine ethyl ester hydrochloride (80 mg, 4 equiv, 0.52 mmol) and DIEA (135 mg, 1.04 mmol, 8 equiv) were dissolved in dry DCM (1 mL), the temperature was cooled to 0°C, and the crude acid chloride was dissolved in DCM and added dropwise to the reaction system. The reaction mixture was reacted at 25°C for 16 h. The reaction was complete as monitored by LCMS. The reaction solution was poured into water and extracted with EA. The EA phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. The product was then purified using a preparative plate (DCM:MeOH = 30:1) to obtain the title compound (20 mg, 88% yield) as a white solid. LCMS (ESI) [M+1] + =658.2. 1H NMR (400MHz, DMSO-d6) δ8.23 (dd, J=1.9, 0.9Hz, 1H), 7.87-7.81 (m, 2H), 4.76 (d, J=13.3Hz,2H),4.52(dd,J=12.8,10.2Hz,1H),4.34(dd,J=12.6,10.4Hz,1H),4.0 8(ttd,J=6.8,4.3,2.0Hz,4H),3.82(dtd,J=17.0,7.1,3.6Hz,2H),3.25(d,J=13 .9Hz,2H),2.04(dd,J=20.6,13.0Hz,3H),1.63-1.55(m,2H),1.27-1.19(m,12H).

[0886] Example 149

[0887] Similarly, using the phosphonate of Example 58 as a starting material, Example 149 was prepared according to steps 1-2 of the preparation method of Example 11. LCMS (ESI): [M+H] + =377.1; 1 H NMR (400MHz, Methanol-d4) δ8.35(s,1H),7.64(d,J=2.4Hz,1H),7.51(d,J=8.8Hz,1H),7.17(dd,J=11.2,2.4Hz,1H),4.56(d,J=12 .8Hz,2H),3.90(s,3H),3.24-3.13(m,2H),2.33-2.17(m,2H),2.15-2.07(m,1H),1.51(dd,J=16.8,6.4Hz,2H),1.45-1.38(m,2H).

[0888] Example 150

[0889] Similarly, starting from Intermediate 28, the dimethyl phosphonate intermediate was obtained according to Steps 1-5 of the preparation method of Example 68, and then Example 150 was prepared according to Steps 1-3 of Example 103. LCMS (ESI): [M+H] + =428.0; 1H NMR (400MHz, DMSO-d6) δ7.97 (d, J=7.2Hz, 1H), 7.51 (s, 1H), 4.68(d,J=12.0Hz,2H),3.35-3.26(m,2H),2.89(d,J=6.0Hz,2H),2.08-2.05(m,3H),1.57(d,J=13.2Hz,2H),1.31-1.22(m,5H).

[0890] Example 151

[0891] Similarly, starting from Intermediate 28, steps 1-5 of the preparation method of Example 68 were followed to obtain dimethyl phosphonate intermediate, and then steps 1-3 of Example 103 were followed to prepare Example 151. LCMS (ESI): [M+H] + =442.0; 1 H NMR(400MHz, Methanol-d4)δ7.59(dd,J=8.0,1.2Hz,1H),7.16-7.10(m,1H),4.83-4.80(m,2H),3.23(t,J=12.0Hz,2H) ,3.15(q,J=6.8Hz,1H),2.19-2.10(m,3H),1.66(dd,J=14.0,6.0Hz,2H),1.46-1.44(m,2H),1.36(s,3H),1.35(s,3H).

[0892] Example 152

[0893] Similarly, starting from Intermediate 28, the dimethyl phosphonate intermediate was obtained according to Steps 1-5 of the preparation method of Example 68, and then Example 152 was prepared according to Steps 1-3 of Example 103. LCMS (ESI): [M+H] + =400.0; 1 H NMR (400MHz, Methanol-d4) δ8.55 (s, 1H), 7.64 (d, J = 6.4Hz, 1H), 7.19-7.14 (m, 1H), 4.83-4.63 (m,2H),3.24-3.21(m,2H),2.21-2.01(m,3H),1.58(dd,J=17.6,6.0Hz,2H),1.46-1.13(m,2H).

[0894] Example 153

[0895] Preparation of (1-(7,9-difluoro-2-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0896] Step 1: Preparation of diethyl (1-(7,9-difluoro-2-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0897] Diethyl (1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (100 mg, 0.21 mmol, 1 equiv) was dissolved in 1,4-dioxane (5 mL) and methanol (0.5 mL). Cesium carbonate (206 mg, 0.63 mmol, 3 equiv), 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (19 mg, 0.04 mmol, 0.2 equiv), and palladium acetate (5 mg, 0.02 mmol, 0.1 equiv) were added. Under argon, the reaction mixture was reacted at 110°C for 2 hours. The reaction mixture was quenched with saturated ammonium chloride and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude title compound (80 mg, yield: 80.8%). LCMS (ESI): [M+H] + =469.1.

[0898] Step 2: Preparation of (1-(7,9-difluoro-2-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0899] Diethyl (1-(7,9-difluoro-2-methoxy-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (80 mg, 0.17 mmol, 1 equivalent) was dissolved in dichloromethane (1 mL), and trimethylsilyl bromide (130 mg, 0.85 mmol, 5 equivalents) was added. The reaction mixture was reacted at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by reverse phase HPLC (C18 column, 10 mol / L ammonia water, acetonitrile) to obtain the title compound (2.24 mg, 0.01 mmol, 3.2%) as a white solid. LCMS (ESI): [M+H] + =413.1; 1H NMR (400MHz, Methanol-d4) δ7.02(dd,J=9.6,2.0Hz,1H),6.61(td,J=10.0,1.6Hz,1H),4.71(d,J=12.8Hz,2 H), 3.96 (s, 3H), 3.20 (t, J = 12.4Hz, 2H), 2.17-2.09 (m, 3H), 1.50 (dd, J = 16.4, 6.4Hz, 2H), 1.41-1.31 (m, 2H).

[0900] Example 154

[0901] Preparation of ((((1-(1-(2-chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonyl)bis(oxy)bis(methylene)bis(2,2-dimethylpropionate):

[0902] (1-(2-Chloro-7,9-difluoro-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid (300 mg, 0.72 mmol, 1 equiv) was dissolved in N,N-dimethylformamide (10 mL). Iodomethyl pivalate (1.74 g, 7.2 mmol, 10 equiv) and N,N'-dicyclohexyl-4-morpholinamidine (420 mg, 1.44 mmol, 2 equiv) were added. The reaction mixture was stirred at 20°C for 16 hours, then heated to 80°C and stirred for 3 hours. The reaction mixture was purified by reverse-phase HPLC (C18 column, acetonitrile, 0.1% formic acid) to obtain the title compound (11 mg, 2.4% yield) as a white solid. LCMS (ESI): [M+H] + =645.4; 1 H NMR(400MHz, Methanol-d4)δ7.09(dd,J=9.2,1.6Hz,1H),6.79(td,J=10.4,2.0Hz,1H),5.74-5.66(m,4 H), 4.57 (d, J = 13.6Hz, 2H), 3.21 (t, J = 11.6Hz, 2H), 2.20-1.93 (m, 3H), 1.52-1.44 (m, 4H), 1.07 (s, 18H).

[0903] Example 155

[0904] Preparation of (1-(7,9-difluoro-2-(methylthio)-5H-pyrimidinyl[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid

[0905] Step 1: Preparation of 7,9-difluoro-2-mercapto-5-pyrimido[5,4-b]indol-4-ol:

[0906] 3-Amino-4,6-difluoro-1H-indole-2-carboxamide (1.0 g, 4.74 mmol, 1 equivalent) was added to anhydrous ethanol (20 mL). Carbon disulfide (1.08 g, 14.2 mmol, 3 equivalents) was added dropwise, followed by solid potassium hydroxide (1.33 g, 23.7 mmol, 5 equivalents). The reaction mixture was heated to 90°C for 2 hours. After cooling, 1N dilute hydrochloric acid was added dropwise to adjust the pH to approximately 6. The precipitated solid was filtered to obtain the title compound (1.10 g, 91.6% yield) as a brownish-red solid. LCMS (ESI): [MH] - =252.0.

[0907] Step 2: Preparation of 7,9-difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indol-4-ol:

[0908] 7,9-Difluoro-2-mercapto-5-pyrimidin[5,4-b]indol-4-ol (300 mg, 1.18 mmol, 1 equivalent) was added to N,N-dimethylformamide (10 mL), followed by anhydrous potassium carbonate (327 mg, 2.37 mmol, 2 equivalents), and iodomethane (168 mg, 1.18 mmol, 1 equivalent) was added dropwise. The mixture was allowed to react at room temperature for 1 hour. The reaction solution was extracted with saturated ammonium chloride solution (10 mL) and ethyl acetate (10 mL x 3), and concentrated to afford the title compound (160 mg, 50.7% yield) as a brown solid. LCMS (ESI): [M+H] + =268.0.

[0909] Step 3: Preparation of 4-chloro-7,9-difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indole

[0910] 7,9-Difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indol-4-ol (80 mg, 0.3 mmol, 1 equivalent) was added to phenylphosphonic acid dichloride (6 mL). The reaction was heated to 180°C for 2 hours. The reaction solution was cooled and diluted with ethyl acetate (10 mL), then neutralized with sodium bicarbonate. The solution was extracted three times with ethyl acetate (20 mL x 3) and concentrated to give the title compound (80 mg, yield: 93.3%) as a yellow solid. LCMS (ESI): [M+H] + =286.0.

[0911] Step 4: Preparation of diethyl (1-(7,9-difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate:

[0912] 4-Chloro-7,9-difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indole (80 mg, 0.28 mmol, 1 eq) and diethyl [(piperidin-4-yl)methyl]phosphonate (132 mg, 0.56 mmol, 2 eq) were added to acetonitrile (10 mL), followed by diisopropylethylamine (0.11 mL, 0.84 mmol, 3 eq). The reaction was heated to 80°C for 1 hour. The mixture was diluted with ethyl acetate (30 mL) and extracted three times with saturated ammonium chloride (20 mL x 3). The mixture was concentrated to afford the title compound (110 mg, 81.1% yield). LCMS (ESI): [M+H] + =485.1.

[0913] Step 5: Preparation of (1-(7,9-difluoro-2-(methylthio)-5H-pyrimidinyl[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonic acid:

[0914] Diethyl (1-(7,9-difluoro-2-(methylthio)-5H-pyrimido[5,4-b]indol-4-yl)piperidin-4-yl)methyl)phosphonate (110 mg, 0.23 mmol, 1 equivalent) was added to dichloromethane (6 ml), followed by trimethylsilyl bromide (0.5 mL, 3.27 mmol, 14.2 equivalents). The reaction was heated to 50°C for 2 hours. After cooling, the reaction solution was concentrated, and the crude product was purified by reverse phase HPLC (C18, 10 mmol / L ammonia water, acetonitrile) to obtain the title compound (14.3 mg, 14.7% yield). LCMS (ESI): [MH] - =427.1; 1 H NMR (400MHz, DMSO-d6) δ7.16(d,J=7.6Hz,1H),6.93(t,J=10.0Hz,1H),4.46(d,J=13.6Hz,2H),3.11 (t,J=12.4Hz,2H),2.36(s,3H),2.11-1.87(m,3H),1.46(dd,J=17.6,6.4Hz,2H),1.36-1.28(m,2H).

[0915] Example 156

[0916] Similarly, using S-1-(3-chlorophenyl)propane-1,3-diol as the starting material, Example 156 was prepared according to Steps 1-2 of the preparation method of Example 144. LCMS (ESI): [M+H] + =567.10; 1 H NMR(400MHz, Methanol-d4)δ:7.46(s,1H),7.42~7.35(m,3H),7.11(dd,J=7.6,1.6Hz,1H),6.81(td,J=10.0,2.0Hz,1H),5.68(d,J=11.2Hz,1H),4.7 4-4.67(m,1H),4.63-4.58(m,1H),4.43?4.34(m,2H),4.27?4.24(m,1H),3 .30?3.18(m,2H),2.33?2.18(m,2H),2.16-1.99(m,5H),1.64?1.42(m,2H).

[0917] Example 157

[0918] Similarly, starting from Intermediate 28, the dimethyl phosphonate intermediate was obtained according to Steps 1-2 of the preparation method of Example 1, and then Example 157 was prepared according to Steps 1-2 of the preparation method of Example 11. LCMS (ESI): [M+H] + =359.9; 11 H NMR (400MHz, DMSO-d6) δ8.53(s,1H),7.95(d,J=7.2Hz,1H),7.39(t,J=9.2Hz,1H),3.56-3.86(m,2H),1.85-1.75(m,2H),1.52-1.43(m,2H).

[0919] Example 158

[0920] Similarly, starting from Intermediate 28, chlorination was carried out according to Step 1 of the preparation method of Example 1, and then the dimethyl phosphonate intermediate was obtained according to Step 4 of the preparation method of Example 87. Example 158 was prepared according to Steps 1-2 of the preparation method of Example 11. LCMS (ESI): [M+H] + =374.0; 1H NMR (400MHz, Methanol-d4) δ8.52 (s, 1H), 7.64 (d, J = 6.4Hz, 1H), 7.19-7.13 (m ,1H),3.91(t,8.0Hz,2H),3.48(s,3H),2.09-1.99(m,2H),1.69-1.61(m,2H).

[0921] Example 159

[0922] Similarly, starting from Intermediate 28, the phosphonic acid diester intermediate was obtained according to Steps 1-2 of the preparation method ...

Claims

1. A compound of formula (I), and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs, in L is selected from -(NR L3 )SO2N(R L1 )(R L2 ),-SO2R L1 、-C(O)OR L1 、-C(O)N(R L1 )(R L2 ),-C(O)NHOR L1 、-P(O)(OR L1 )(OR L2 )、-P(O)(NHR L1 )(OR L2 )、-P(O)(OR L1 )(R L2 )、-P(O)(NHR L1 )(NHR L2 )、-P(O)(NHR L1 )(R L2 )、-P(O)(R L1 )(R L2 )、-P(O)(SR L1 )(SR L2 )、-P(O)(NHR L1 )(SR L2 )、-P(O)(SR L1 )(R L2 ) or -B(OH)2, where R L1 、R L2 and R L3 R is independently selected from hydrogen, deuterium, halogen, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, or optionally substituted aryl; or R L1 and R L2 are linked to form an optionally substituted ring structure; A is selected from optionally substituted alkylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, optionally substituted cycloalkylenealkylene, optionally substituted heterocyclylenealkylene, optionally substituted arylenealkylene, or optionally substituted heteroarylenealkylene; Z is selected from (chemical bond), -O-, -S-, -CO-, -SO-, -SO2-, -C(R Z1 )(R Z2 )-、-N(R Z1 )-、-CH2N(R Z1 )-、-N(R Z1 )CH2-、-CON(R Z1 )-、-N(R Z1 )CO-、-SON(R Z1 )-、-SO2N(R Z1 )-、-N(R Z1 )SO-、-N(R Z1 )SO2- or where R Z1 and R Z2 is independently selected from hydrogen, deuterium, halogen, hydroxy, thiol, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl; Y is selected from -N(R Y )-, -O- or -S-, where R Y is selected from hydrogen, deuterium, optionally substituted alkyl or optionally substituted cycloalkyl; n = 0, 1, 2 or 3; R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 、-NR 3 COR 4 、-COR 3 、-OCOR 3 、-SONR 3 R 4 、-SO2NR 3 R 4 、 -NR 3 SOR 4 、-NR 3 SO2R 4 or -Si(R 3 )(R 4 )(R 5 ), R 2 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR 3 、-SR 3 、-NR 3 R 4 、COOR 3 、-CONR 3 R 4 or-COR 3 , where R 3 、R 4 or R 5 independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; X 1 and X 2 independently selected from N or C(R X ), where R X are independently selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, -OR a 、-SR a 、-NR a R b 、-COOR a 、-CONR a R b 、-NR b COR a 、-COR a 、-OCOR a 、-SONR a R b 、-SO2NR a R b 、-NR b SOR a 、-NR b SO2R a or -Si(R a )(R b )(R c ), where R a 、R b and R c R is independently selected from hydrogen, deuterium, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; or R 2 With X 1 or X 2 are linked to form an optionally substituted ring structure.

2. The compound according to claim 1, wherein R L1 、R L2 and R L3 are independently selected from hydrogen, deuterium, halogen, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group or optionally substituted C 6-10 Aryl.

3. The compound according to claim 1, wherein R L1 、R L2 and R L3 independently selected from hydrogen, deuterium, halogen, amino, cyano; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl and aryl; and alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl and aryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclyl (e.g., 4-6 membered heterocyclyl), -OC(O)-alkyl (e.g., -OC(O)-C 1-6 alkyl), -C(O)O-alkyl (e.g. -C(O)OC 1-6 Alkyl) or -OC(O)O-alkyl (e.g. -OC(O)OC 1-6 alkyl); 4. The compound according to claim 1, wherein R L1 、R L2 and R L3 are independently selected from hydrogen, deuterium, amino, optionally substituted alkyl (e.g., C 1-6 alkyl) and optionally substituted aryl (e.g., C 6-10 aryl); Preferably, R L1 、R L2 and R L3 independently selected from hydrogen, deuterium, amino, unsubstituted alkyl and phenyl, and alkyl substituted with one or more substituents independently selected from the group consisting of deuterium, -OC(O)-C 1-6 Alkyl, -C(O)OC 1-6 Alkyl, -OC(O)OC 1-6 alkyl.

5. The compound according to claim 1, wherein L is selected from: -(NR L3 )SO2N(R L1 )(R L2 ),-SO2R L1 、 -C(O)OR L1 、-C(O)NHOR L1 、-P(O)(OR L1 )(OR L2 )、-P(O)(OR L1 )(R L2 )、-P(O)(NHR L1 )(NHR L2 )、-P(O)(NHR L1 )(OR L2 ) or -B(OH)2; Preferably, L is selected from: -P(O)(OH)2, -C(O)OH, -C(O)NHOH, -P(O)(OH)(OC2H5), -P(O)(OC2H5)2, -SO2NH2, -NHSO2NH2, -P(O)(OH)(CH3), -P(O)(OCH2OC(O)C(CH3)3)2, -P(O)(NHCH(CH3)C(O)OC2H5)2, -P(O)(NHCH(CH3)C(O)OCH(CH3)2)2, -P(O)(OCH2OC(O)OCH(CH3)2)2, -P(O)(OH)(OCH2OC(O)C(CH3)3), -P(O)(OH)(OC6H5), -P(O)(NHCH(CH3)C(O)OCH(CH3)2)(OC6H5).

6. The compound according to claim 1, wherein R L1 and R L2 connected to form an optionally substituted 5- or 6-membered ring structure; Preferably, R L1 and R L2 connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl, optionally substituted 4-6 membered heterocyclyl, optionally substituted phenyl or optionally substituted 5-6 membered heteroaryl; More preferably, L is -P(O)(OR L1 )(OR L2 ), R L1 and R L2 are independently optionally substituted alkyl, and R L1 and R L2 They are linked to form an optionally substituted 5- or 6-membered ring structure.

7. The compound according to claim 1, wherein A is selected from optionally substituted C 1-6 Alkylene, optionally substituted C 3-6 Cycloalkylene, optionally substituted 4-10 membered heterocyclylene, optionally substituted C 6-10 Arylene, optionally substituted 5-10 membered heteroarylene, optionally substituted C 3-6 Cycloalkylene C 1-3 Alkylene, optionally substituted 4-10 membered heterocyclylene C 1-3 Alkylene, optionally substituted C 6-10 Arylene C 1-3 Alkylene or optionally substituted 5-10 membered heteroarylene C 1-3 Alkylene.

8. The compound of claim 1, wherein A is selected from unsubstituted alkylene, cycloalkylene, heterocyclylene, arylene, heteroarylene, cycloalkylenealkylene, heterocyclylenealkylene, arylenealkylene, or heteroarylenealkylene; and alkylene, cycloalkylene, heterocyclylene, arylene, heteroarylene, cycloalkylenealkylene, heterocyclylenealkylene, arylenealkylene, or heteroarylenealkylene substituted with one or more substituents independently selected from deuterium, halogen, hydroxyl, sulfhydryl, amino, cyano, acetyl, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclic group (e.g. 4-6 membered heterocyclic group), -O-alkyl (e.g. -OC 1-6 Alkyl), -NH-alkyl (e.g. -NHC 1-6 Alkyl), -S-alkyl (e.g. -SC 1-6 Alkyl), -O-cycloalkyl (e.g. -OC 1-6 Cycloalkyl), -NH-cycloalkyl (e.g., -NHC 1-6 Cycloalkyl) or -S-cycloalkyl (e.g. -SC 1-6 cycloalkyl).

9. The compound of claim 1, wherein A is selected from methylene, ethylene, propylene, butylene; optionally substituted cyclohexylene, piperidinylene, piperazinylene, diazacycloheptylene, bicyclo[1.1.1]pentylene, phenylene, furanylene, cyclohexylenealkylene, piperidinylenealkylene, piperazinylenealkylene, diazacycloheptylenealkylene, bicyclo[1.1.1]pentylenealkylene, phenylenealkylene, or furanylenealkylene; Preferably, A is selected from methylene, ethylene, propylene, butylene; unsubstituted or substituted by alkyl (e.g. C 1-6 alkyl, such as methyl) substituted cyclohexylene, piperidinylene, piperazinylene, diazepanylene, bicyclo[1.1.1]pentylene, phenylene or furylene; and unsubstituted or substituted alkyl (e.g. C 1-6 alkyl, such as methyl) substituted cyclohexylene C 1-2 Alkylene, piperidinylene C 1-2 Alkylene, piperazinyl C 1-2 Alkylene, diazepanyl C 1-2 Alkylene, bicyclo[1.1.1]pentyl C 1-2 Alkylene, phenylene C 1-2 Alkylene or furylene C 1-2 Alkylene.

10. The compound according to claim 1, wherein R Z1 and R Z2 are independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl or an optionally substituted 4- to 6-membered heterocyclic group.

11. The compound according to claim 1, wherein R Z1 and R Z2 independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl; and alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from deuterium, halogen, hydroxyl, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 cycloalkyl) or a heterocyclic group (e.g., a 4- to 6-membered heterocyclic group).

12. The compound according to claim 1, wherein Z is selected from -(chemical bond), -N(R Z1 )-or-N(R Z1 )CH2-; Preferably, R Z1 and R Z2 are independently selected from hydrogen, deuterium and optionally substituted alkyl (e.g., C 1-6 alkyl, such as methyl); More preferably, Z is selected from (chemical bond), -NH-, -N(CH3)- or -NHCH2-.

13. The compound according to claim 1, wherein R Y is selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl or optionally substituted C 3-6 Cycloalkyl.

14. The compound according to claim 1, wherein R Y is selected from hydrogen; deuterium; unsubstituted alkyl or cycloalkyl; and alkyl or cycloalkyl substituted with one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, mercapto, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), -OC(O)-alkyl (e.g. -OC(O)-C 1-6 Alkyl) or -C(O)O-alkyl (e.g. -C(O)OC 1-6 alkyl).

15. The compound according to claim 1, wherein R Y Selected from hydrogen, deuterium, unsubstituted or replaced by -OC(O)-C 1-6 Alkyl substituted C 1-6 alkyl; Preferably, Y is selected from -NH-, -N(CH3)-, -N(CH2OC(O)C(CH3)3)-, -O- or -S-.

16. The compound according to claim 1, wherein R 1 Selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, nitro, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 Aryl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 、-NR 3 COR 4 、-COR 3 、-OCOR 3 、-SONR 3 R 4 、-SO2NR 3 R 4 、-NR 3 SOR 4 、-NR 3 SO2R 4 or -Si(R 3 )(R 4 )(R 5 ).

17. The compound according to claim 1, wherein R 1 is selected from unsubstituted alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl; and alkyl, alkenyl, cycloalkyl, heterocyclyl or aryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

18. The compound according to claim 1, wherein R 2 is selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclyl, -OR 3 、-SR 3 、-NR 3 R 4 、-COOR 3 、-CONR 3 R 4 or-COR 3 .

19. The compound according to claim 1, wherein R 2 is selected from unsubstituted alkyl, alkenyl, cycloalkyl or heterocyclyl; and alkyl, alkenyl, cycloalkyl or heterocyclyl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

20. The compound according to claim 1, wherein R 3 、R 4 and R 5 are independently selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 aryl or optionally substituted 5-6 membered heteroaryl.

21. The compound according to claim 1, wherein R 3 、R 4 and R 5 independently selected from hydrogen; deuterium; unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; and alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl substituted with one or more substituents independently selected from deuterium, halogen, hydroxy, thiol, amino, cyano, nitro, carboxyl, optionally substituted alkyl, optionally substituted cycloalkyl or optionally substituted alkoxy.

22. The compound according to claim 1, wherein R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, amino, cyano; unsubstituted or halogen-substituted alkyl, alkenyl, cycloalkyl or aryl (e.g. phenyl); -OR 3 、-NR 3 R 4 、-COOR 3 or -NR 3 COR 4 ; Preferably, R 3 and R 4 independently selected from hydrogen; deuterium; alkyl groups which are unsubstituted or substituted with halogen (e.g., C 1-6 alkyl) or cycloalkyl (e.g. C 3-6 Cycloalkyl) More preferably, R 1 Selected from halogen, hydroxy, amino, cyano, -CF3, ethyl, vinyl, cyclopropyl, phenyl substituted with -F, -OCH3, -OCH2CH3, -COOH, -NHCOCH3, -NH-cyclopentyl, -NH(CH3)2.

23. The compound according to claim 1, wherein R 2 Selected from hydrogen, deuterium, halogen, hydroxyl; alkyl, alkenyl or cycloalkyl which is unsubstituted or substituted by deuterium or halogen; -NR 3 R 4 、-OR 3 or -SR 3 ; Preferably, R 2 Selected from halogen, hydroxy, methyl, -CF3, -CHD2, ethyl, propyl, isopropyl, propenyl, cyclopropyl, -NHCH3, -SCH3 or -OCH3; 24. The compound according to claim 1, wherein R X are independently selected from hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, nitro, optionally substituted C 1-6 Alkyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclyl, -OR a 、-SR a 、-NR a R b 、-COOR a 、-CONR a R b 、-NR b COR a 、-COR a 、-OCOR a 、-SONR a R b 、-SO2NR a R b 、-NR b SOR a 、-NR b SO2R a or -Si(R a )(R b )(R c ), where R a 、R b and R c are independently selected from hydrogen, deuterium, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 Cycloalkyl, optionally substituted 4-6 membered heterocyclic group, optionally substituted C 6-10 aryl or optionally substituted 5-6 membered heteroaryl; and R X The substituents in the optional substitution are selected from one or more of the following: hydrogen, deuterium, halogen, hydroxyl, thiol, amino, cyano, alkyl (e.g., C 1-6 Alkyl), alkenyl (e.g. C 2-6 alkenyl), alkynyl (e.g. C 2-6 Alkynyl), cycloalkyl (e.g. C 3-6 Cycloalkyl), heterocyclic group (e.g. 4-6 membered heterocyclic group), aryl group (e.g. C 6-10 aryl) or heteroaryl (e.g., 5-6 membered heteroaryl).

25. The compound according to claim 1, wherein X 1 and X 2 are independently selected from N or CH.

26. The compound according to claim 1, wherein R 2 With X 1 or X 2 connected to form an optionally substituted 5- or 6-membered ring structure; Preferably, R 2 With X 1 or X 2 connected to form a cyclic structure, which is further substituted by one or more substituents independently selected from the group consisting of deuterium, halogen, hydroxyl, thiol, amino, cyano, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-6 cycloalkyl, optionally substituted 4- to 6-membered heterocyclyl, optionally substituted aryl, or optionally substituted 5- to 6-membered heteroaryl.

27. The compound of claim 1, wherein the compound is selected from:

28. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 27 and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs as an active ingredient.

29. The pharmaceutical composition of claim 28, further comprising additional drugs (such as small molecules, peptides, nucleic acids, antibodies, antibody-drug conjugates) for preventing and / or treating cancer or tumors, and / or additional therapeutic agents for treating inflammatory diseases, autoimmune diseases, and immune-mediated diseases.

30. The pharmaceutical composition of claim 29, wherein the additional drugs for preventing and / or treating cancer or tumors include, but are not limited to, cell signaling inhibitors, chlorambucil, melphalan, cyclophosphamide, ifosfamide, busulfan, carmustine, lomustine, streptozotocin, cisplatin, carboplatin, oxaliplatin, dacarbazine, temozolomide, procarbazine, methotrexate, fluorouracil, cytarabine, gemcitabine, mercaptopurine, fludarabine, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, topotecan, irinotecan, etoposide, trabectedin, dactinomycin, doxorubicin, epirubicin , daunorubicin, mitoxantrone, bleomycin, mitomycin C, ixabepilone, tamoxifen, flutamide, gonadorelin analogs, megestrol acetate, prednisone, dexamethasone, methylprednisolone, thalidomide, interferon alfa, leucovorin, sirolimus, temsirolimus, everolimus, afatinib, alisertib, amuvatinib, apatinib, axitinib, bortezomib, bosutinib, brivanib, cabozantinib, cediranib, crenolanib, crizotinib, dabrafenib, dacomitinib, danusetib, dasatinib, dovitinib, erlotinib, foretinib, gane tespib, gefitinib, ibrutinib, icotinib, imatinib, iniparib, lapatinib, lenvatinib, linifanib, linsitinib, masitinib, momelotinib, motesanib, neratinib, nilotinib, niraparib, oprozomib, olaparib, pazopanib, pictilisib, ponatinib, quizartinib, regorafenib, rigosertib, rucaparib, ruxolitinib, sacatinib, saridegib, sorafenib, sunitinib, telatinib, tivantinib, tivozanib , tofacitinib, trametinib, vandetanib, veliparib, vemurafenib, vismodegib, volasertib, alemtuzumab, bevacizumab, berentuzumab vedotin, catumaxomab, cetuximab, denosumab, gemtuzumab, ipilimumab, nimotuzumab, ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, PI3K inhibitor, CSF1R inhibitor, A2A and / or A2B receptor antagonist, IDO inhibitor, anti-PD-1 antibody, anti-PD-L1 antibody, LAG3 antibody, TIM-3 antibody, TIGIT antibody, CD47 antibody, CLAUDIN 18.2 antibody, anti-CTLA-4 antibody, or any combination thereof.

31. The pharmaceutical composition according to claim 29, wherein the antibody-drug conjugate includes but is not limited to: trastuzumab enroute, brentuximab, olgaituzumab, vedicituzumab, gosartan, delutec-trastuzumab, or any combination thereof.

32. The pharmaceutical composition of claim 29, wherein the additional therapeutic agent for treating inflammatory diseases, autoimmune diseases and immune-mediated diseases is selected from steroidal drugs (e.g., prednisone, hydroprednisolone, methylhydroprednisolone, cortisone, hydroxycortisone, betamethasone, dexamethasone, etc.), methotrexate, leflunomide, anti-TNFα agents (e.g., etanercept, infliximab, adalimumab, etc.), calcineurin inhibitors (e.g., tacrolimus, pimecrolimus, etc.), antihistamines (e.g., diphenhydramine, hydroxyzine, loratadine, ebastine, ketotifen, cetirizine, levocetirizine, fexofenadine, etc.) or any combination thereof.

33. Use of a compound of formula (I) according to any one of claims 1 to 27 and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs, or a pharmaceutical composition according to any one of claims 28 to 32 in the preparation of a medicament for preventing and / or treating a disease mediated by ENPP1.

34. The use according to claim 33, wherein the disease mediated by ENPP1 is selected from cancer, tumor, inflammatory disease, autoimmune disease, neurodegenerative disease, attention-related disease or immune-mediated disease.

35. The use according to claim 34, wherein the cancer and tumor include: Skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis carcinoma, hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, testicular cancer, urinary cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumor, Hodgkin lymphoma, non-Hodgkin lymphoma Lymphoma, Burkitt lymphoma, leukemia, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder cancer, bronchogenic carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myeloma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmacytoma.

36. The method of claim 34, wherein the inflammatory diseases, autoimmune diseases and immune-mediated diseases include: Arthritis, rheumatoid arthritis, spondyloarthritis, gouty arthritis, osteoarthritis, juvenile arthritis, other arthritic conditions, lupus, systemic lupus erythematosus (SLE), skin-related diseases, psoriasis, eczema, dermatitis, atopic dermatitis, pain, lung disease, lung inflammation, adult respiratory distress syndrome (ARDS), pulmonary sarcoidosis, chronic inflammatory lung disease, chronic obstructive pulmonary disease (COPD), cardiovascular disease, atherosclerosis, myocardial infarction, congestive heart failure, myocardial ischemia-reperfusion injury, inflammatory bowel disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, asthma, Sjögren's syndrome, autoimmune thyroid disease, urticaria (wheal), multiple sclerosis, scleroderma, organ transplant rejection, xenotransplantation, idiopathic thrombocytopenic purpura (ITP), Parkinson's disease, Alzheimer's disease, diabetes-related diseases, inflammation, pelvic inflammatory disease, allergic rhinitis, allergic bronchitis and allergic sinusitis.

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