Fused tricyclic PARP1 inhibitors, methods for their preparation, and uses

Abstract

The present invention relates to fused tricyclic PARP1 inhibitors, their preparation methods, and uses. Specifically, the present invention relates to compounds that inhibit poly(ADP-ribose) polymerase activity, pharmaceutical compositions thereof, and uses thereof. The present invention particularly relates to compounds of formula (I), pharmaceutical compositions containing these compounds, and the use of these compounds in the preparation of medicaments for preventing and / or treating diseases, particularly diseases ameliorated by inhibition of PARP1. [Formula 1] TIFF2025538192000118.tif42170

Description

[Technical Field]

[0001] The present invention relates to compounds having poly(ADP-ribose) polymerase 1 (PARP1) inhibitory activity and uses thereof, particularly to quinolone compounds, pharmaceutical compositions containing said compounds, and uses thereof in drugs for diseases, particularly tumor diseases, that are ameliorated by inhibiting PARP1. [Background technology]

[0002] Poly(ADP-ribose) polymerase (PARP) is a novel enzyme family capable of catalyzing the transfer of ADP-ribose to target proteins (poly(ADP-ribosylation)). At least 18 PARP family members are encoded by different genes and share homology in a conserved catalytic domain (Non-Patent Document 1). The full name of PARP1 is poly(ADP-ribose) polymerase 1. PARP1 is an abundant nuclear protein (Non-Patent Document 2). PARP1 can catalyze the transfer of ADP-ribose residues from NAD+ to target substrate proteins or nucleic acids, building poly(ADP-ribose) (PAR) chains and adding them to downstream target proteins. This post-translational modification is called PARylation. PARPs play an important role in several cellular processes, including cell proliferation and cell death (Non-Patent Document 2). The primary function of PARP is to participate in the repair of DNA damage. The most common damage is a DNA single-strand break (SSB), which can potentially cause resection and be converted into a lethal DNA double-strand break (DSB). PARP1 binds to damaged DNA at single-strand DNA breaks (SSBs) and other DNA damage sites. This event induces a series of conformational changes in the structure of PARP1, activating its catalytic function (Non-Patent Document 3).

[0003] The BRCA1 and BRCA2 proteins are essential for the repair of double-stranded DNA breaks (DSBs), a repair process called homologous recombination repair (HRR). This is a form of DNA repair that uses homologous DNA sequences to guide repair at DSBs (Non-Patent Document 3). HRR is generally a "conservative" mechanism because it usually restores the original DNA sequence at the site of DNA damage. When cells have HRR defects, whether caused by defects in BRCA1, BRCA2, or other pathway components, non-conservative forms of DNA repair, such as non-homologous end joining (NHEJ), become dominant.

[0004] PARP inhibitors exert their anticancer effects by inhibiting DNA damage repair in highly mutated cancer cells, causing "toxic damage" and resulting in the death of cells with defective homologous recombination repair (HRR). Healthy cells have multiple signaling pathways for DNA repair, so inhibiting PARP alone is not particularly toxic to cells. On the other hand, in some tumor cells, specific genetic mutations, such as BRCA, disrupt other DNA repair pathways, making the DNA repair pathway dependent on PARP-1 and therefore particularly sensitive to PARP inhibitors. Therefore, patients with ovarian and breast cancers carrying BRCA mutations are more likely to benefit from PARP inhibitors. PARP2 content is low, accounting for only 5%–10% of total PARP activity. Knocking out PARP1 significantly reduces PARP activity compared with knocking out PARP2 (<10%) (Non-Patent Document 4). Knocking out PARP1 blocks the inhibitory activity of olaparib against PARP and also eliminates its cell proliferation inhibitory effect (Non-Patent Document 5). These data indicate that PARP1 is the key PARP that determines the efficacy of PARPi. Corresponding literature reports indicate that intact PARP2 in the bone marrow is required for mouse survival. Lack of PARP2 leads to reduced numbers of RBCs, WBCs, and BM cells (Non-Patent Document 6). Compared with PARP1 knockout, PARP2 knockout can reduce the number of T cells and RBCs, but PARP1 knockout has no apparent effect on the number of T cells (Non-Patent Document 4) or RBCs (Non-Patent Document 7). Therefore, inhibition of PARP1 is the primary cause of efficacy, while inhibition of PARP2 is the primary cause of toxicity. The development of highly selective PARP1 / 2 inhibitors is expected to significantly reduce PARP2-induced toxicity without significantly reducing efficacy. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Morales et al,Critical Reviews in Eukaryotic Gene Expression 24.1,2014 [Non-patent document 2] Murai et al, Cancer Research 72.21, 2012 [Non-patent document 3] Lord et al,Science 355.6330,2017 [Non-patent document 4] Yelamos et al,The EMBO Journal 25.18,2006 [Non-patent document 5] Murai et al, Cancer Research, 2012 [Non-patent document 6] Farres et al, Blood, The Journal of the American Society of Hematology 122.1, 2013 [Non-Patent Document 7] Farres et al,Cell Death & Differentiation 22.7,2015 Summary of the Invention [Problem to be solved by the invention]

[0006] As described above, there is an urgent need in the art to develop PARP inhibitors with high efficacy and favorable safety, especially inhibitors with high selectivity for PARP1. [Means for solving the problem]

[0007] The present invention relates to compounds represented by general formula (I), or stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, crystalline forms, solvates, hydrates or prodrugs thereof. [ka] (where, X1 are independently -N-, -NR 14 -, -CR 7 -, -CR 7 R 7’ , -CH2CR 7 R 7’ -, -CR 7 R 7’ selected from —CH—, O, and S; X 2 are independently -N-, -NR 15 -, -CR 8 -, -CR 8 R 8’ -, O, and S; X 3 are independently -N-, -NR 16 -, -CR 9 -, O, and S; X 4 , X 8 are each independently selected from -N-, and -C-; [ka] is a single bond or a double bond, and X 1 , X 2 , X 3 , X 4 , X 8 together form a 5-membered heteroaryl, or a partially saturated 5- or 6-membered heterocyclyl, wherein said heteroaryl or heterocyclyl each independently contains 1, 2 or 3 heteroatoms independently selected from N, O or S; R 7 , R 7’ , R 8 , R 8’ , R 9 are each independently selected from hydrogen, halogen, hydroxy, cyano, C1-C3 alkoxy, unsubstituted or substituted C3-C6 cycloalkyl, or unsubstituted or substituted C1-C6 alkyl; or R 7 and R 7’ or R 8 and R 8’ together form a C3-C6 cycloalkyl, and R7 , R 7’ , R 8 , R 8’ , R 9 are each independently preferably hydrogen, halogen or C1-C4 alkyl, and R 7 , R 7’ , R 8 , R 8’ , R 9 are each independently more preferably hydrogen, F or methyl, or preferably R 7 and R 7’ or R 8 and R 8’ together form a C3-C4 cycloalkyl, such as cyclopropyl or cyclopentyl; R 14 , R 15 , R 16 are each independently selected from hydrogen, unsubstituted or substituted C3-C6 cycloalkyl, and unsubstituted or substituted C1-C6 alkyl; R 14 , R 15 , R 16 are each independently preferably hydrogen or C1-C3 alkyl, and R 14 , R 15 , R 16 are each independently more preferably methyl; X 5 , X 6 are each independently -N- and -CR 10 - selected from R 10 is selected from hydrogen, halogen, cyano, unsubstituted or substituted C1-C6 alkoxy, and unsubstituted or substituted C1-C6 alkyl; R 10 is preferably hydrogen, halogen, cyano, or C1-C4 alkyl, and R 10 is more preferably hydrogen, fluorine, chlorine or methyl; X 7 -N- or -CR 17 - and R 17 is selected from hydrogen, halogen, cyano, unsubstituted or substituted C1-C6 alkoxy, and unsubstituted or substituted C1-C6 alkyl; R 17is preferably hydrogen, halogen, cyano, or C1-C4 alkyl, and R 17 is more preferably hydrogen, fluorine, chlorine or methyl, R 1 , R 1 ', R 2 , R 3 , R 4 , R 5 are each independently selected from hydrogen, unsubstituted or substituted C1-C6 alkyl, or R 4 , R 5 together with the carbon atom to which they are attached, form a C3-C6 cycloalkyl; s and n are each independently selected from 0, 1, and 2; Y is N or CH; R 6 teeth, [ka] is selected from Each R 11 are independently selected from halogen, cyano, C1-C3 alkoxy, carbonyl, -CONHR 13 , amino, preferably halogen, -CONHR 13 and cyano, more preferably -CONHR 13 is selected from m is 0, 1, 2, or 3; R 12 is selected from hydrogen, halogen, cyano, and unsubstituted or substituted C1-C4 alkyl; R 13 is hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C3-C8 cycloalkyl, unsubstituted or substituted C1-C6 alkoxy, or unsubstituted or substituted 3- to 8-membered heterocycloalkyl, preferably R 13 is hydrogen, unsubstituted or halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, or C1-C6 alkoxy, and the heterocycloalkyl refers to a heterocycloalkyl containing 1 to 3 heteroatoms selected from N, O, and S, and preferably R 13is methyl, ethyl, C2-C3 alkoxy, cyclopropanyl, propylene oxide, oxetanyl, or oxiranyl; where R 1 , R 1’ , R 2 , R 3 , R 4 , R 5 , R 7 , R 7’ , R 8 , R 8’ , R 9 , R 10 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 The substitution in refers to substitution with one or more selected from C1-C4 alkyl, halogen, hydroxy, cyano, amino, carboxy, and C3-C6 cycloalkyl; however, X 5 and X 8 If both are -N-, then X 1 , X 2 , X 3 , X 4 at least one of is -N-; X 4 and X 5 If both are -N-, then X 3 is also -N-, X 3 is oxygen and X 1 , X 2 , X 4 , X 8 is -C- and X 7 Ga-CR 17 -If R 1 , R 1’ , R 17 is not hydrogen at the same time, Preferably, however, X 8 If is -N-, then X 1 , X 2 , X 3 At least one of is -N-.

[0008] Preferably, however, X 5 , X 6 are each independently -CR 10 - and X 7 -CR 17 -It is.

[0009] More preferably, however, X 8 If is -N-, then X 1 , X 2 , X 3 at least one of is -N-; X 4 and X 5 If both are -N-, then X 3 is also -N-, X 3 is oxygen and X 1 , X 2 , X 4 , X 8 is -C- and X 7 Ga-CR 17 -If R 1 , R 1’ , R 17 is not hydrogen at the same time.

[0010] In one preferred embodiment of the present invention, there is provided a compound represented by general formula (II), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof: [ka] (where, X 1 are independently -N-, -NR 14 -, -CR 7 -, O, and S; X 2 are independently -N-, -NR 15 -, and -CR 8 -, O, and S; X 3 are independently -N-, -NR 16 -, and -CR 9-, O, and S; X 4 , X 8 are each independently selected from -N-, and -C-; [ka] is a single bond or a double bond, and X 1 , X 2 , X 3 , X 4 , X 8 together form a 5-membered heteroaryl or a partially saturated 5-membered heterocyclyl; R 7 , R 8 , R 9 are each independently selected from hydrogen, halogen, unsubstituted or substituted C3-C6 cycloalkyl, or unsubstituted or substituted C1-C6 alkyl; R 7 , R 8 , R 9 The substitution in R refers to substitution with one or more selected from C1-C4 alkyl, halogen, hydroxy, cyano, amino, carboxy, and C3-C6 cycloalkyl; 7 , R 8 , R 9 is preferably hydrogen, halogen, or C1-C4 alkyl, and R 7 , R 8 , R 9 is more preferably hydrogen, F, or methyl; X 5 , X 6 , X 7 , R 1 , R 1 ', R 2 , R 3 , R 4 , R 5 , R 6 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , s, n, m, and Y are as defined in general formula (I).

[0011] In one preferred embodiment of the present invention, in the compounds represented by general formula (I) and (II), X 1 , X 2 , X 3 , X 4 , X 8 together form a partially saturated 5-membered heterocyclyl or a partially saturated 6-membered heterocyclyl. 4 If C, then X 3 is N or -NR 16 More preferably, X 4 If C, then X 3 is N.

[0012] In one preferred embodiment of the present invention, in the compounds represented by general formula (I) and (II), X 1 , X 2 , X 3 , X 4 , X 8 together form a 5-membered heteroaryl.

[0013] In one preferred embodiment of the present invention, in the compounds represented by general formula (I) and (II), X 1 , X 2 , X 3 , X 4 , X 8 together form a partially saturated 5-membered heterocyclyl.

[0014] In one preferred embodiment of the present invention, in the compounds represented by general formula (I) and (II), X 1 , X 2 , X 3 , X 4 , X 8 together form a partially saturated 6-membered heterocyclyl.

[0015] In one preferred embodiment of the present invention, in the compounds represented by general formula (I) and (II), X 1 , X 2 , X 3 , X 4, X 8 At least one of X is nitrogen, and 1 , X 2 , X 3 , X 4 , X 8 together constitute a partially saturated 5- or 6-membered heterocyclyl.

[0016] In one preferred embodiment of the present invention, in the compound represented by general formula (I) or (II), X 1 are independently -N-, -NR 14 -, -CR 7 -, -CR 7 R 7’ , -CH2CR 7 R 7 '-, and -CR 7 R 7’ -CH2-, X 2 are independently -N-, -NR 15 -, -CR 8 -, and -CR 8 R 8’ - selected from X 4 , X 8 are each independently selected from -N-, and -C-; X 3 are independently -N-, -NR 16- , and -CR 9 - selected from, and X 1 , X 2 , X 3 , X 4 , X 8 At least one of X is nitrogen, and 1 , X 2 , X 3 , X 4 , X 8 together constitute a partially saturated 5- or 6-membered heterocyclyl.

[0017] In one preferred embodiment of the present invention, in the compound represented by general formula (I) or (II),

[0018] X 1 are independently -CR 7 -, -CR 7 R 7’ , -CH2CR 7 R 7’ -, and -CR 7 R 7’ -CH2-,

[0019] X 2 are independently -CR 8 -, and -CR 8 R 8’ - selected from

[0020] X 4 is -C-,

[0021] X 8 is -N-,

[0022] X 3 -N-, and -NR 16 is selected from

[0023] And X 1 , X 2 , X 3 , X 4 , X 8 together constitute a partially saturated 5- or 6-membered heterocyclyl.

[0024] In the above embodiment, the term "partially saturated 5- or 6-membered heterocyclyl" refers to a heterocyclyl that is unsaturated but not aromatic.

[0025] Preferably, in the general formula (I) or (II), [ka] is selected from the following structures: [ka]

[0026] More preferably, it is selected from the following structures: [ka] TIFF2025538192000010.tif48170

[0027] Here, in the general formula (I) or (II), [ka] is preferably selected from the following structures: [ka]

[0028] More preferably, it is selected from the following structures: [ka]

[0029] More preferably, it is selected from the following structures: [ka] (However, R 2 , R 3 , R 4 , R 5 , R 11 , R 12 , R 13 , m are defined as above.)

[0030] More preferably, said compound of formula (I) is selected from the following specific compounds: [Table 1] TIFF2025538192000016.tif251170TIFF2025538192000017.tif239170TIFF2025538192000018.tif248170TIFF2025538192000019.tif230170

[0031] In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate, or prodrug thereof, and a pharmaceutically acceptable carrier.

[0032] In certain embodiments of the pharmaceutical composition, the pharmaceutical composition is formulated for intravenous, intramuscular, oral, rectal, inhalation, nasal, topical, ocular, or otic administration. In other embodiments of the pharmaceutical composition, the pharmaceutical composition is a tablet, pill, capsule, liquid, inhalant, nasal spray solution, suppository, solution, emulsion, ointment, eye drops, or ear drops. In other embodiments of the pharmaceutical composition, the pharmaceutical composition further comprises one or more additional therapeutic agents.

[0033] In another aspect, the present invention provides the use of a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, or said pharmaceutical composition, in the preparation of a medicament for the prevention or treatment of a disease ameliorated by inhibiting PARP1.

[0034] In another aspect, the present invention provides a method for preventing or treating a disease ameliorated by inhibiting PARP1, comprising administering to an individual in need of such treatment an effective amount of a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, or said pharmaceutical composition.

[0035] In some embodiments of the present invention, the disease includes, but is not limited to, cancer.

[0036] In some embodiments of the present invention, the cancer includes, but is not limited to, malignant tumors such as ovarian cancer, breast cancer, fallopian tube cancer, endometrial cancer, peritoneal cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, biliary tract cancer, osteosarcoma, cervical cancer, head and neck tumors, germ cell cancer and embryonal carcinoma, esophageal cancer, malignant glioma, Ewing's sarcoma, pancreatic cancer, melanoma, bile duct cancer, prostate cancer, small cell lung cancer, non-small cell lung cancer, lymphoma, and blood cancer.

[0037] In some embodiments of the invention, the cancer genome is homologous recombination repair deficient.

[0038] In some embodiments of the invention, the cancer is dependent on DNA double strand breaks and defective homologous recombination repair pathways.

[0039] In some embodiments of the invention, the cancer comprises one or more cancer cells, which are deficient in the ability to repair DNA double-strand breaks by homologous recombination compared to normal cells.

[0040] In some embodiments of the invention, the cancer comprises one or more cancer cells, wherein the cancer cells are BRCA1 or BRCA2 deficient or have a BRCA1 or BRCA2 mutation.

[0041] Terminology In the present invention, unless otherwise specified, the terms used in the present invention have the meanings defined below. Terms not expressly defined herein have the general meanings commonly understood by those skilled in the art.

[0042] As used herein, the term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

[0043] As used herein, "heteroaryl" refers to a monocyclic ring system having 5 to 6 (5-6-membered) or 6 (6-membered) ring atoms, an aromatic ring containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S, with the remaining ring atoms being carbon atoms. For example, it may contain one N heteroatom and, optionally, one, two, or three heteroatoms independently selected from N, O, or S. If the total number of S and O atoms in a heteroaryl exceeds 1, these S and O heteroatoms are not adjacent to each other. For example, the heteroaryl includes, but is not limited to, a 5- to 6-membered monocyclic heteroaryl, i.e., a monocyclic aromatic hydrocarbon group having 5 or 6 ring atoms and containing 1, 2, or 3 heteroatoms independently selected from N, O, or S, with the remaining ring atoms being carbon atoms. Examples of said heteroaryl include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl, thiazolyl, isothiazolyl, pyrazinyl, pyridazinyl, pyridinyl, or pyrimidinyl.

[0044] As used herein, "heterocyclyl" refers to a fully saturated or partially saturated non-aromatic monocyclic, bicyclic, or tricyclic ring group having 3 to 15 ring atoms (e.g., 4 to 12 ring atoms, 3 to 10 ring atoms, 5 to 10 ring atoms, 4 to 7 ring atoms, 5 to 6 ring atoms), such as a 4- to 7-membered monocyclic, 5- to 6-membered monocyclic ring system containing 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S, with the remaining ring atoms being carbon atoms. The nitrogen and sulfur heteroatoms may optionally be oxidized, e.g., the sulfur heteroatom may form an -S(O)- or -S(O)2- structure. The heterocyclyl may be a monocyclic, bicyclic, fused, spiro, or bridged ring group. "5- to 6-membered heterocyclyl" means a heterocyclyl having 5 or 6 ring atoms, which includes 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S, e.g., 1, 2, or 3 N heteroatoms, and is a monocyclic ring.

[0045] As used herein, "heterocycloalkyl" refers to a fully saturated heterocyclyl as defined herein. For example, a "3- to 8-membered heterocycloalkyl" refers to a saturated heterocycle having 3 to 8 ring atoms, including 1, 2, 3, or 4 heteroatoms independently selected from N, O, or S, e.g., including one N or O heteroatom.

[0046] As used herein, the terms "optional," "any," or "optionally" mean that the subsequently described substitution pattern, event, or circumstance may or may not occur, and the description includes cases where said substitution pattern occurs and cases where said substitution pattern does not occur. For example, "optionally substituted alkyl" includes "unsubstituted alkyl" and "substituted alkyl," as defined herein. One of ordinary skill in the art will understand that for any group containing one or more substituents, said group does not include substitution patterns that are sterically impractical, chemically incorrect, synthetically infeasible, and / or inherently unstable.

[0047] As used herein, the term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of the present invention and are not biologically or otherwise undesirable. Non-limiting examples of such salts include non-toxic inorganic or organic base or acid addition salts of the compounds of the present invention. In many cases, the compounds of the present invention are capable of forming acid salts and / or base salts due to the presence of amino and / or carboxy or similar groups. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Particularly preferred are ammonium, potassium, sodium, calcium, and magnesium salts. Organic bases from which salts can be derived include, for example, primary amines, secondary amines, tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, particularly, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound (basic or acidic moiety) by conventional chemical methods. Generally, the salts can be prepared by reacting the free acid form of the compound with a stoichiometric amount of an appropriate base (e.g., hydroxide, carbonate, bicarbonate, etc. of Na, Ca, Mg, or K), or by reacting the free base form of the compound with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water, an organic solvent, or a mixture of the two.Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred, where practicable. Other suitable salts are described in Remington's Pharmaceutical Sciences, 20th Edition, Mack Publishing Company, Easton, Pa., (1985), which is incorporated herein by reference.

[0048] As used herein, the term "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drugs, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, and similar substances, and combinations thereof, that are well known to those skilled in the art (see, e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing Company, 1990, pp. 1289-1329, which is incorporated herein by reference). Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the therapeutic or pharmaceutical compositions is contemplated.

[0049] As used herein, the term "solvate" is meant to include stoichiometric and non-stoichiometric solvent addition forms. When the solvent is water, the solvate formed is a hydrate, and when the solvent is ethanol, the solvate formed is an ethanolate. Hydrates are formed by one or more water molecules and one molecule of the substance, where the water retains its molecular state as HO, and such combinations can form one or more types of hydrates, such as hemihydrates, monohydrates, and dihydrates.

[0050] As used herein, a "prodrug" refers to a chemically modified active or inactive compound that, after administration to an individual, undergoes physiological action in vivo (e.g., hydrolysis, metabolism, etc.) to become a compound of the present invention. The suitability and techniques for the preparation and use of prodrugs are well known to those skilled in the art.

[0051] The term "therapeutically effective amount" of a compound of the invention refers to an amount of a compound of the invention that is capable of eliciting a biological or medical response, ameliorating symptoms, slowing the progression of a disease, or preventing a disease in an individual.

[0052] As used herein, the term "individual" refers to an animal. Preferably, the animal is a mammal. An individual also refers to, for example, a primate (e.g., a human), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, fish, bird, etc. In a preferred embodiment, the individual is a human.

[0053] As used herein, the term "inhibition" refers to the reduction or inhibition of a particular disease, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0054] As used herein, in one embodiment, the term "treating" a so-called disease or disorder refers to ameliorating the disease or disorder (i.e., arresting or slowing the progression of the disease or at least one clinical symptom thereof). In another embodiment, "treatment" refers to improving at least one physical parameter that may not be noticeable to the patient. In another embodiment, "treatment" refers to modulating the disease or disorder physically (e.g., stable observable symptoms) or physiologically (e.g., stabilization of a physical parameter), or both. [Effects of the Invention]

[0055] The main advantages of the present invention are that the compounds of the present invention have high selectivity for PARP1, have fewer side effects than olaparib (AZD-2281), and have high clinical application value. DETAILED DESCRIPTION OF THE INVENTION

[0056] Unless otherwise noted, the experimental materials and reagents used in the following examples are commercially available. The raw materials are generally commercially available or readily prepared by methods known to those skilled in the art.

[0057] In each example, the experimental equipment and materials used are as follows:

[0058] 1 H NMR was recorded on a Varian Mercury-300 or Varian Mercury-400 nuclear magnetic resonance spectrometer. 13 C NMR was recorded on a Varian Mercury-400, Varian Mercury-500, or Varian Mercury-600 nuclear magnetic resonance spectrometer; chemical shifts are expressed in ppm. Mass spectra were recorded on a Finnigan / MAT-95 (EI), Finnigan LCQ / DECA, and Micromass Ultra Q-TOF (ESI) mass spectrometer. Silica gel used for reversed-phase preparative HPLC separation was 200–300 mesh.

[0059] Abbreviation [Table 2]

[0060] [Synthesis of key intermediates] Intermediate 1a: (R)-6-chloro-N-methyl-5-(2-methylpiperazin-1-yl)picolinamide hydrochloride [ka]

[0061] Step 1: Synthesis of methyl 5-bromo-6-fluoropicolinate Compound 1a-1 (1 g, 4.6 mmol), acetonitrile (30 mL), and silver difluoride (1.76 g, 13.9 mmol) were added sequentially to a 50 mL single-neck flask and stirred overnight. The reaction mixture was filtered, and the filtrate was concentrated and purified using a silica gel column (PE:EA = 5:1) to give 1a-2 (450 mg, white solid, yield: 42%). LCMS (ESI): m / z 233.9 [M+H] + ;RT=1.51min (3.00min).

[0062] Step 2: Synthesis of t-butyl (R)-4-(2-chloro-6-(methoxycarbonyl)pyridin-3-yl)-3-methylpiperazine-1-carboxylate A 50 mL single-neck flask was charged with 1a-2 (450 mg, 1.9 mmol), (R)-4-Boc-2-methylpiperazine (577 mg, 2.9 mmol), Ruphos Pd G3 (159 mg, 0.19 mmol), cesium carbonate (1.5 g, 4.7 mmol), and dioxane (6 mL). The mixture was heated overnight at 80 °C under nitrogen protection. The reaction mixture was concentrated and purified using a silica gel column (PE:EA = 2:1) to give product 1a-3 (300 mg, yellow solid, 44% yield). LCMS (ESI): m / z 354.1 [M+H] + ;RT=1.80min(3.00min).

[0063] Step 3: t-Butyl (R)-4-(2-chloro-6-(methylcarbamoyl)pyridin-3-yl)-3-methylpiperazine-1-carboxylate 1a-3 (100 mg, 0.28 mmol) and methylamine ethanol solution (2 mL) were added to a 50 mL single-neck flask, and the reaction mixture was stirred overnight. Concentration gave 1a-4 (80 mg, yellow solid, yield: 80%). LCMS (ESI): m / z 297.1 [M-56+H] + ;RT=1.70min(3.00min).

[0064] Step 4: Synthesis of (R)-6-chloro-N-methyl-5-(2-methylpiperazin-1-yl)picolinamide hydrochloride A 20 mL single-neck flask was charged with 1a-4 (80 mg, 0.23 mmol), EA (2 mL), and 4 M hydrochloric acid in dioxane (2 mL), and the mixture was stirred at room temperature for 2 hours. Concentration gave 1a (65 mg, yellow oil, 100% yield). LCMS (ESI): m / z 253.2 [M+H] + ;RT=0.99min(3.00min).

[0065] Intermediate 2a: Synthesis of N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride [ka]

[0066] Step 1: Synthesis of t-butyl 4-(2-chloro-6-(methoxycarbonyl)pyridin-3-yl)piperazine-1-carboxylate A 50 mL flask was charged with 1a-2 (500 mg, 2.1 mmol), 1-t-butoxycarbonylpiperazine (600 mg, 3.2 mmol), Ruphos Pd G3 (180 mg, 0.21 mmol), cesium carbonate (1.7 g, 5.2 mmol), and dioxane (15 mL). The mixture was heated overnight at 80 °C under nitrogen protection. The reaction mixture was concentrated and purified using a silica gel column (PE:EA = 2:1) to give 2a-1 (640 mg, yellow solid, 88% yield). LCMS (ESI): m / z 340.1 [M+H] + ;RT=1.74min(3.00min).

[0067] Step 2: Synthesis of 5-(4-(t-butoxycarbonyl)piperazin-1-yl)-6-fluoropicolinic acid 2a-1 (320 mg, 0.94 mmol) and THF (8 mL) were added to a dry 50 mL single-neck flask, and lithium hydroxide monohydrate (200 mg, 4.7 mmol) in water (8 mL) was added dropwise. The mixture was stirred for 2 hours, and the pH of the reaction mixture was adjusted to 6 with 1 M hydrochloric acid. The reaction mixture was concentrated and purified using a reverse-phase column (2% to 40% acetonitrile in water) to give 2a-2 (300 mg, yellow solid, 92% yield). LCMS (ESI): m / z 326.1 [M+H] + ;RT=1.26min(3.00min).

[0068] Step 3: Synthesis of t-butyl 4-(2-chloro-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate Compound 2a-2 (300 mg, 0.92 mmol), 1-hydroxybenzotriazole (149 mg, 1.1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (211 mg, 1.1 mmol), DIEA (237 mg, 1.84 mmol), DMF (6 mL), and methylamine hydrochloride (123 mg, 1.84 mmol) were added to a dry 50 mL single-neck flask and stirred overnight at room temperature. Water (20 mL) was added to the reaction mixture, which was then extracted three times with EA (30 mL). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered under vacuum. The filtrate was concentrated and purified using a reverse-phase column (20%-70% acetonitrile in water) to give 2a-3 (250 mg, white solid, 80% yield). LCMS (ESI): m / z 283.1 [M-100+H] + ;RT=1.66min(3.00min).

[0069] Step 4: Synthesis of 6-chloro-N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride A dry 20 mL single-neck flask was charged with 2a-3 (250 mg, 0.74 mmol), EA (3 mL), and 4 M HCl in dioxane (1 mL) in that order, stirred at room temperature for 2 h, and concentrated to give product 2a (200 mg, yellow solid), 100% yield. LCMS (ESI): m / z 239.1 [M+H] +;RT=0.91min (3.00min). 1 H NMR (600MHz, CD3OD): 7.98-7.96 (m, 1H), 7.67-7.64 (m, 1H), 3.50-3.48 (m, 4H), 3.45-3.43 (m, 4H), 2.93 (d, J = 3.6Hz, 3H).

[0070] Intermediate 3a: Synthesis of N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride [ka]

[0071] The synthesis method was the same as that of Intermediate 1a, except that in Step 2 of the preparation method of Intermediate 1a, Boc-piperazine was used instead of (R)-4-Boc-2-methylpiperazine, and Compound 1a-1 was used instead of Compound 1a-2. LCMS (ESI): m / z 221.2 [M+H] + ;RT=0.285min(6.00min).

[0072] Intermediate 5a: Methyl (6-chloro-4-oxo-4,5-dihydropyrrolo[1,2-a]quinoxalin-7-yl)methanesulfonate [ka]

[0073] Step 1: Synthesis of 1-bromo-2,4-dichloro-3-nitrobenzene A 250 mL single-neck flask was charged with 5a-1 (25.0 g, 157 mmol) and 100 mL of concentrated sulfuric acid. N-bromosuccinimide (33.6 g, 188 mmol) was slowly added in an ice bath and the reaction was allowed to proceed at 80°C overnight. The reaction mixture was diluted by slowly pouring ice water into the reaction mixture, extracted twice with EA, dried, concentrated, and then subjected to column chromatography (PE) to obtain product 5a-2 (36 g, yellow oil, yield: 96.6%). 1 H NMR (400MHz, DMSO-d6): δ8.19-8.14 (m, 1H), 7.56-7.51 (m, 1H).

[0074] Step 2: Synthesis of methyl 1-(4-bromo-3-chloro-2-nitrophenyl)-1H-pyrrole-2-carboxylate 5a-2 (0.5 g, 2.1 mmol), methyl 1H-pyrrole-2-carboxylate (244 mg, 1.89 mmol), and cesium carbonate (1.37 g, 4.2 mmol) were added to a 100 mL single-neck flask containing 15 mL of DMF. The mixture was allowed to react at room temperature for 5 h. The reaction mixture was diluted with water, extracted twice with EA, dried, concentrated, and then purified by column chromatography (PE:EA = 5:1) to give 5a-3 (4.0 g, yellow solid, 65.5% yield). LCMS (ESI): m / z 345.0 [M+H] + ;RT=1.783min(2.50min).

[0075] Step 3: Synthesis of 7-bromo-6-fluoropyrrolo[1,2-a]quinoxalin-4(5H)-one 5a-3 (6.5 g, 18.9 mmol) and iron powder (21 g, 37.9 mmol) were added to a single-neck flask containing 250 mL of acetic acid, and the mixture was reacted at 110°C for 3 h. EA was added to the reaction mixture, which was then filtered. The filtrate was concentrated, and saturated sodium bicarbonate solution was added. The mixture was extracted three times with EA. The organic phase was washed with water, dried, and concentrated to give 5a-4 (650 mg, yellow solid, yield: 12.2%). 1 H NMR (400MHz, DMSO-d6): δ11.46(s,1H),8.22(m,1H),7.89(d,J=8.0Hz,1H),7.53-7.48(m,1H),7.10(d,J=3.2Hz,1H),6.74-6.73(m,1H).

[0076] Step 4: Synthesis of 6-chloro-7-(hydroxymethyl)pyrrole[1,2-a]quinoxalin-4(5H)-one 5a-4 (700 mg, 2.49 mmol), XPhos Pd G2 (197 mg, 0.25 mmol), and (tri-t-butylstannyl)methanol (960 mg, 2.99 mmol) were added to a three-neck flask containing 15 mL of anhydrous dioxane and reacted overnight at 100 °C. The reaction mixture was quenched by adding potassium fluoride solution, filtered, and the filtrate was concentrated to give 5a-5 (430 mg, yellow solid, yield: 74.4%). LCMS (ESI): m / z 233.1 [M+H] + ;RT=0.877min(2.00min). 1 H NMR (400MHz, DMSO-d6): δ11.23(s,1H),8.18-8.17(m,1H),7.87(d,J=8.4Hz,1H),7.28-7. 24(m,1H),7.08-7.07(s,1H),6.72-6.70(m,1H),5.36-5.33(m,1H),4.59(d,J=5.6Hz,2H).

[0077] Step 5: Synthesis of methyl (6-chloro-4-oxo-4,5-dihydropyrrolo[1,2-a]quinoxalin-7-yl) methanesulfonate 5a-5 (150 mg, 0.65 mmol) and triethylamine (164 mg, 1.63 mmol) were added to a 100 mL three-neck flask containing 8 mL of THF. Methanesulfonyl chloride (89 mg, 0.78 mmol) was added in an ice bath, and the mixture was allowed to react at room temperature for 3 h. The reaction mixture was concentrated to give 5a (120 mg, yellow solid). LCMS (ESI): m / z 309.1 [M−H] - ;RT=1.241min(2.50min).

[0078] Intermediate 6a: 7-(1-bromoethyl)pyrrolo[1,2-a]quinoxalin-4(5H)-one [ka]

[0079] Step 1: Synthesis of methyl 1-(4-bromo-2-nitrophenyl)-1H-pyrrole-2-carboxylate 6a-1 (500 mg, 2.27 mmol), methyl 1H-pyrrole-2-carboxylate (341.26 mg, 2.73 mmol), and cesium carbonate (1.48 g, 4.55 mmol) were dissolved in DMF (10 mL), and the reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was diluted with water (20 mL), extracted with EA (20 mL × 3), dried, concentrated, and then subjected to column chromatography (PE:EA = 10:1) to give 6a-2 (383 mg, 1.18 mmol, yield: 51.9%). LCMS (ESI): m / z 325.0 [M]; RT = 1.341 min (2.50 min).

[0080] Step 2: Synthesis of 7-bromopyrrolo[1,2-a]quinoxalin-4(5H)-one 6a-2 (1.5 g, 4.61 mmol) and iron powder (5.15 g, 92.27 mmol) were dissolved in acetic acid (50 mL) and stirred at 110 °C for 3 h. Concentration gave a dark brown solid, which was diluted with DCM. The organic phase was washed with saturated aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 6a-3 (1.3 g, 4.49 mmol, yield: 100%, yellow solid). 1 H NMR (400MHz, DMSO-d6): δ11.35(s,1H),8.20(s,1H),8.03(d,J=8.4Hz,1H),7. 45(s,1H),7.37(d,J=8.4Hz,1H),7.06(d,J=3.6Hz,1H),6.71(d,J=2.8Hz,1H).

[0081] Step 3: Synthesis of 7-acetylpyrrolo[1,2-a]quinoxalin-4(5H)-one 6a-3 (500 mg, 1.9 mmol), tributyl(1-ethoxyvinyl)tin (1.37 g, 3.8 mmol), and PdCl2(PhP3)2 (200 mg, 0.29 mmol) were dissolved in 1,4-dioxane (25 mL) and reacted at 100 °C for 16 h. After cooling to room temperature, 1 M aqueous hydrochloric acid (10 mL) was added to the reaction solution and stirred for 10 min. The mixture was extracted with EA (10 mL × 3), dried, concentrated, and then subjected to column chromatography (THF:PE = 1:1) to give crude product 6a-4 (220 mg, 0.97 mmol, yield: 51%). LCMS (ESI): m / z 227 [M+H] + ;RT=0.947(2.50min).

[0082] Step 4: Synthesis of 7-(1-hydroxyethyl)pyrrolo[1,2-a]quinoxalin-4(5H)-one Sodium borohydride (73.58 mg, 1.94 mmol) was slowly added to a solution of 6a-4 (220 mg, 0.97 mmol) in methanol (5 mL) at 0 °C, and the mixture was allowed to react at room temperature for 2 h. The mixture was quenched with saturated aqueous ammonium chloride (5 mL), extracted with EA, dried, concentrated, and then subjected to column chromatography (DCM:methanol = 1:1) to give 6a-5 (200 mg, 0.88 mmol, yield: 90.3%, white solid). LCMS (ESI): m / z 229.1 [M−H] - ;RT=0.877min(2.50min). 1 H NMR(400MHz,DMSO-d6):δ11.23(s,1H),8.15(s,1H),7.98(d,J=8.4Hz,1H),7.33(s,1H),7.16(d,J=8.4Hz,1H) ,7.02(d,J=3.6Hz,1H),6.67(t,J=3.2Hz,1H),5.30(d,J=4.0Hz,1H),4.78-4.75(m,1H),1.35(d,J=6.4Hz,3H).

[0083] Step 5: Synthesis of 7-(1-bromoethyl)pyrrolo[1,2-a]quinoxalin-4(5H)-one Phosphorus tribromide (711.55 mg, 2.63 mmol) was slowly added to a solution of 6a-5 (200 mg, 0.87 mmol) in DCM (5 mL) at 0 °C, and the mixture was allowed to react at room temperature for 2 h. The reaction mixture was quenched with water, extracted with EA, dried, and concentrated to give crude product 6a (200 mg, 0.69 mmol, yield: 79.0%, white solid). LCMS (ESI): m / z 289.1 [M−H] - ;RT=1.447(2.50min).

[0084] Intermediate 7a: 7-(1-bromomethyl)-6-Fpyrazolo[1,2-a]quinoxalin-4(5H)-one [ka]

[0085] The synthetic method is referenced to Intermediate 6a, except that 5a-4 prepared in Step 3 of Intermediate 5a was used instead of 6a-3 used in Step 4 of Intermediate 6a. LCMS (ESI): m / z 310.0 [M+H] + ;RT=1.202min(2.50min).

[0086] Intermediate 8a: 7-(Bromomethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one [ka]

[0087] Step 1: Synthesis of N-(5-bromo-2-fluorophenyl)-1H-pyrazole-5-carboxamide 8a-1 (3.0 g, 15.8 mmol), 1H-pyrazole-5-carboxylic acid (1.95 g, 17.4 mmol), DIEA (6.1 g, 47.4 mmol), and HATU (9.0 g, 23.7 mmol) were added to a single-neck flask containing 30 mL of DMF and reacted at room temperature for 16 h. The reaction mixture was diluted with water, extracted with EA, washed with saturated sodium chloride solution, dried, concentrated, and then purified by column chromatography (PE:EA = 5:1) to give 8a-2 (2.0 g, yellow solid, 44.8% yield). LCMS (ESI): m / z 286.0 [M+H] + ;RT=1.03min(2.00min).

[0088] Step 2: Synthesis of 7-bromopyrazol[1,5-a]quinoxalin-4(5H)-one 8a-2 (2.0 g, 7.07 mmol) was added to a three-neck flask containing 30 mL of DMAC, and sodium hydride (565 mg, 14.1 mmol) was added in an ice bath. The reaction mixture was then reacted at 145 °C for 16 h. The reaction mixture was quenched with saturated ammonium chloride solution, and the solid precipitated was filtered and dried to give 8a-3 (1.5 g, yellow solid, yield: 80.7%). LCMS (ESI): m / z 264.0 / 266.0 [M+H] + ;RT=0.93min(2.00min). 1 H NMR (400MHz, DMSO-d6): δ11.50 (s, 1H), 8.03-7.97 (m, 2H), 7.47-7.39 (m, 2H), 7.10 (s, 1H).

[0089] Step 3: Synthesis of 7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one 8a-3 (500 mg, 1.90 mmol), (tributylstannyl)methanol (671 mg, 2.09 mmol), and XPhos Pd G2 (75 mg, 0.095 mmol) were added to a single-neck flask containing 10 mL of dioxane. The mixture was reacted at 80 °C for 16 h under nitrogen protection. The reaction mixture was concentrated and then purified by column chromatography (DCM:methanol = 15:1) to give product 8a-4 (190 mg, yellow solid). Yield: 45.0%. LCMS (ESI): m / z 214.1 [M−H] - ;RT=0.984min(2.50min).

[0090] Step 4: Synthesis of 7-(bromomethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one 8a-4 (80 mg, 0.37 mmol), triphenylphosphine (293 mg, 1.12 mmol), and carbon tetrabromide (248 mg, 0.74 mmol) were added to a 100 mL single-neck flask containing 8 mL of DCM, and the mixture was allowed to react at room temperature for 2 hours. The reaction mixture was concentrated to give crude product 8a (80 mg, yellow solid). LCMS (ESI): m / z 278.0 [M+H] + ;RT=1.405min(2.50min).

[0091] Intermediate 9a: 7-(1-bromoethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one [ka]

[0092] The synthetic method is referenced to Intermediate 6a, except that 8a-3 prepared in Step 2 of Intermediate 8a was used instead of 6a-3 used in Step 4 of Intermediate 6a. LCMS (ESI): m / z 290.0 [M−H] - ;RT=1.300min(2.50min).

[0093] Intermediate 10a: Synthesis of methyl (6-chloro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxalin-7-yl)methanesulfonate [ka]

[0094] Step 1: Synthesis of N-(2,6-difluorophenyl)acetamide To a single-neck flask, 10a-1 (2.00 g, 15.49 mmol), DCM (20 mL), and acetic anhydride (1.53 mL, 16.27 mmol) were added sequentially. The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with water (20 mL) and adjusted to pH 8 with saturated aqueous sodium bicarbonate solution, resulting in the precipitation of a white solid. The filter cake was collected by filtration to give 10a-2 (2.60 g, white solid, yield: 98.07%). LCMS (ESI): m / z 172.2 [M+H] + ;RT=1.036min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ9.69 (s, 1H), 7.37-7.30 (m, 1H), 7.17-7.13 (m, 2H), 2.08 (s, 3H).

[0095] Step 2: Synthesis of N-(3-bromo-2,6-difluorophenyl)acetamide 10a-2 (1.00 g, 5.84 mmol), concentrated sulfuric acid (10 mL), and N-bromosuccinimide (1.04 g, 5.84 mmol) were added sequentially to a single-neck flask at room temperature. The mixture was stirred at room temperature and allowed to react for 16 hours. The reaction mixture was poured into ice water (50 mL), and a white solid precipitated. The filter cake was collected by vacuum filtration to give 10a-3 (1.26 g, white solid, yield: 86.24%). LCMS (ESI): m / z 291.0 [M+H+MeCN] + ;RT=1.290min(2.50min).

[0096] Step 3: Synthesis of 3-bromo-2,6-difluoroaniline 10a-3 (1.00 g, 4.00 mmol), ethanol (6 mL), and concentrated hydrochloric acid (3 mL) were added to a single-neck flask at room temperature, followed by sequential addition. The mixture was heated to 70°C and reacted for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was adjusted to pH 8 with saturated aqueous sodium bicarbonate and extracted with EA. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 10a-4 (650 mg, yellow solid, yield: 78.14%). LCMS (ESI): m / z 207.9 [M+H+MeCN] + ;RT=1.636min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ6.93-6.88 (m, 1H), 6.81-6.76 (m, 1H), 5.54 (s, 2H).

[0097] Step 4: Synthesis of N-(3-bromo-2,6-difluorophenyl)-1H-pyrazole-5-carboxamide 1H-Pyrazole-5-carboxylic acid (3.23 g, 28.85 mmol), anhydrous DCM (40 mL), oxalyl chloride (2.28 mL, 26.92 mmol), and DMF (0.10 mL) were added sequentially to a dry 100 mL three-neck flask at 0 °C. The reaction was allowed to proceed at room temperature for 3 h, and then 10a-4 (2.00 g, 9.62 mmol) was slowly added to a solution of 10a-4 (2.00 g, 9.62 mmol) in DCM (30 mL) and pyridine (15 mL) at 0 °C. The mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated aqueous ammonium chloride (100 mL) and extracted with DCM (80 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography (PE:EA = 5:1) to give 10a-5 (1.80 g, near-white solid, yield: 61.97%). LCMS(ESI):m / z303.9[M+H] + ;RT=1.287min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ13.49(s,1H),10.01(s,1H),7.92(d,J=1.6Hz,1H),7.75-7.69(m,1H),7.26-7.21(m,1H),6.77(t,J=2.0Hz,1H).

[0098] Step 5: Synthesis of 7-bromo-6-fluoropyrazolo[1,5-a]quinoxalin-4(5H)-one Sodium hydride (60% wt, 185 mg, 4.63 mmol), DMAC (5 mL), and a solution of 10a-5 (700 mg, 2.32 mmol) in DMAC (5 mL) were added sequentially to a dry 50 mL three-neck flask at 0 °C. The mixture was heated to 145 °C and reacted for 16 h. The mixture was diluted with saturated aqueous ammonium chloride (50 mL) to precipitate and filtered. The filtrate was extracted with EA (15 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was slurried with EA (10 mL) and filtered. The filter cake was collected to give crude product 10a-6 (180 mg, yellow solid). LCMS (ESI): m / z 282.0 [M+H] + ;RT=1.470min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ12.08(s,1H),8.13(d,J=2.0Hz,1H),7.89(dd,J1=1.6Hz,J2=9.6Hz,1H),7.59-7.56(m,1H),7.22(d,J=2.0Hz,1H).

[0099] Step 6: Synthesis of 6-chloro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxalin-4(5H)-one A dry 25 mL three-neck flask was charged with 10a-6 (180 mg, 0.64 mmol), 1,4-dioxane (5 mL), and XPhos Pd G2 (50 mg, 0.06 mmol) sequentially at room temperature. Under nitrogen protection, (tributylstannyl)methanol (246 mg, 0.77 mmol) was added. The mixture was heated to 80 °C and reacted for 16 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was slurried with EA (10 mL) and filtered to obtain compound 10a-7 (48 mg, yellow solid, yield: 32.25%). LCMS (ESI): m / z 234.1 [M+H]+; RT = 0.998 min (2.50 min).

[0100] Step 7: Synthesis of methyl (6-chloro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxalin-7-yl) methanesulfonate A 25 mL one-neck flask was charged with 10a-7 (48 mg, 0.21 mmol), THF (5 mL), triethylamine (0.09 mL, 0.62 mmol), and methanesulfonyl chloride (0.02 mL, 0.25 mmol) in that order. The reaction was allowed to proceed at room temperature for 1 h. The mixture was concentrated under reduced pressure. The mixture was diluted with water (10 mL) and extracted with EA. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 10a (58 mg, yellow solid). LCMS (ESI): m / z 312.0 [M+H] + ;RT=1.223min(2.50min).

[0101] Intermediate 11a: Synthesis of 7-(bromomethyl)-6-chloro-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0102] Step 1: Synthesis of 3-bromo-2-chloro-6-iodobenzoic acid 11a-1 (4.0 g, 13.3 mmol) was sequentially added to a three-neck flask containing 40 mL of THF at room temperature, followed by lithium diisopropylamide (7.3 mL, 14.6 mmol) at -78 °C. The mixture was reacted for 1 hour, then warmed to room temperature, and reacted overnight. The reaction mixture was quenched with saturated ammonium chloride, diluted with water, adjusted to pH 8-9 with dilute sodium hydroxide solution, extracted twice with EA, adjusted to acidic pH with dilute hydrochloric acid, and extracted twice with EA. The combined organic phase was washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 11a-2 (3.2 g, yellow solid, yield: 69.8%). 1 H NMR (400MHz, CDCl3): δ7.68 (d, J=8.4Hz, 1H), 7.57-7.53 (m, 1H).

[0103] Step 2: Synthesis of t-butyl (3-bromo-2-chloro-6-iodophenyl)carbamate 11a-2 (3.0 g, 8.7 mmol), diphenylphosphoryl azide (2.87 g, 10.4 mmol), and triethylamine (1.05 g, 10.4 mmol) were added to a three-neck flask containing 40 mL of toluene. The mixture was allowed to react at 120 °C for 1 hour, after which t-butanol was added and the reaction was continued for 3 hours. The reaction mixture was concentrated and then subjected to column chromatography (PE / EA = 30 / 1) to give 11a-3 (2.9 mg, white solid). LCMS (ESI): m / z 413.8 [M−H] - ;RT=1.684min(2.50min).

[0104] Step 3: Synthesis of t-butyl (3-bromo-2-chloro-6-(1-methyl-1H-pyrazol-5-yl)phenyl)carbamate A flask containing 15 mL of dioxane and 5 mL of water was charged with 11a-3 (1.4 g, 3.37 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (631 mg, 3.04 mmol), potassium carbonate (1.40 g, 10.1 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (catalytic amount). The reaction mixture was stirred at 70 °C for 3 h. The reaction mixture was cooled to room temperature, diluted with water, and extracted twice with EA. The combined organic phases were washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (EA:PE = 1:10) to give 11a-4 (900 mg, yellow solid, yield: 72.3%). LCMS(ESI):m / z370.0[M+H] + ;RT=1.444min(2.50min).

[0105] Step 4: Synthesis of 3-bromo-2-chloro-6-(1-methyl-1H-pyrazol-5-yl)aniline 11a-4 (900 mg, 2.44 mmol) was added to a 50 mL single-neck flask containing 15 mL of methanolic hydrochloric acid. The mixture was allowed to react at room temperature for 3 hours. The solvent was removed, the mixture was diluted with water, the pH was adjusted to alkaline with saturated sodium bicarbonate solution, and the mixture was extracted twice with EA. The combined organic phases were washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 11a-5 (580 mg, yellow solid, yield: 88.4%). LCMS (ESI): m / z 270.3 / 272.3 [M+H] + ;RT=1.22min(2.00min).

[0106] Step 5: Synthesis of 7-bromo-6-chloro-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 11a-5 (780 mg, 2.9 mmol) and carbonyldiimidazole (1.41 g, 8.70 mmol) were added to a 50 mL single-neck flask containing 15 mL of DMF, and the mixture was allowed to react overnight at 170 °C. The reaction mixture was diluted with water, and the solid precipitated. The solid was filtered and dried to give 11a-6 (650 mg, brown solid, yield: 76.0%). LCMS (ESI): m / z 296.2 [M+H] + ;RT=0.85min(2.00min). 1 H NMR (400MHz, DMSO-d6): δ11.51 (s, 1H), 8.15 (s, 1H), 7.97 (d, J = 8.8Hz, 1H), 7.57-7.53 (m, 1H), 4.35 (s, 3H).

[0107] Step 6: Synthesis of 6-chloro-7-(hydroxymethyl)-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 11a-6 (200 mg, 0.68 mmol), (tributylstannyl)methanol (239 mg, 0.75 mmol), and XPhos Pd G2 (27 mg, 0.03 mmol) were added to a single-neck flask containing 10 mL of dioxane, and the mixture was allowed to react at 80 °C for 16 hours. The reaction mixture was concentrated, slurried with EA, and filtered to give 11a-7 (140 mg, yellow solid, yield: 83.8%). LCMS (ESI): m / z 248.2 [M+H]+ ;RT=0.895min (2.50min).

[0108] Step 7: Synthesis of 7-(bromomethyl)-6-chloro-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 11a-7 (70 mg, 0.28 mmol), triphenylphosphine (222 mg, 0.85 mmol), and carbon tetrabromide (188 mg, 0.57 mmol) were added to a 100 mL single-neck flask containing 8 mL of DCM, and the mixture was allowed to react at room temperature for 2 hours. The reaction mixture was concentrated to give crude product 11a (50 mg, yellow solid). LCMS (ESI): m / z 310.3 [M+H] + ;RT=0.64min(2.00min).

[0109] Intermediate 12a: N-methyl-5-(pyrrolidin-3-yloxy)picolinamide hydrochloride [ka]

[0110] Step 1: Synthesis of methyl 5-((1-(t-butoxycarbonyl)pyrrolidin-3-yl)oxy)picolinate Sodium hydride (641 mg, 16 mmol) was added to a dry 250 mL three-neck flask, and under nitrogen protection, THF (40 mL), 12a-1 (2 g, 10.7 mmol), and 12a-2 (1.66 g, 10.7 mmol) were added in an ice bath. The mixture was allowed to react at room temperature for 1 hour. The reaction mixture was extracted with EA (30 mL x 3). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 3:1) to give 12a-3 (1 g, yellow oil, yield: 29%). LCMS (ESI): m / z 323.2 [M+H] + ;RT=1.59min(2.50min).

[0111] Step 2: t-Butyl 3-((6-(methylcarbamoyl)pyridin-3-yl)oxy)pyrrolidine-1-carboxylate A dry 100 mL one-neck flask was charged with 12a-3 (400 mg, 1.24 mmol) and a 30% ethanol solution of methylamine (6 mL). The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with water (10 mL) and extracted with EA (10 mL x 3). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 12a-4 (380 mg, yellow oil). LCMS (ESI): m / z 266 [M+H] + ;RT=1.560min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ8.57(d,J=4Hz,1H),8.28(d,J=2.4Hz,1H),7.98(d,J=8.8Hz,1H),7.58-7.56(m,1H),5.18(s,1H) ),4.88(d,J=3.2Hz,1H),3.44-3.40(m,3H),3.11(s,1H),2.79(d,J=4.4Hz,3H),1.40(d,J=5.6Hz,9H),1.19-1.56(m,1H).

[0112] Step 3: Synthesis of N-methyl-5-(pyrrolidin-3-yloxy)picolinamide hydrochloride 12a-4 (380 mg, 1.18 mmol), DCM (6 mL), and a solution of hydrochloric acid in dioxane (4.0 M, 6 mL) were added to a dry 100 mL single-neck flask in an ice bath. The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to give 12a (300 mg, crude yellow solid). LCMS (ESI): m / z 220 [M+H] + ;RT=0.603min (2.5min).

[0113] Intermediate 13a: Synthesis of 7-(1-bromoethyl)-6-fluoropyrazolo[1,5-a]quinoxalin-4(5H)-one [ka]

[0114] The synthetic method is referenced to Intermediate 6a, except that 10a-6 prepared in Step 5 of Intermediate 10a was used instead of 6a-3 used in Step 4 of Intermediate 6a. LCMS (ESI): m / z 310.0 [M−H] - ;RT=1.548min(2.50min).

[0115] Intermediate 14a: Synthesis of 7-(1-bromoethyl)-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0116] Step 1: Synthesis of 5-bromo-2-(1-methyl-1H-pyrazol-5-yl)aniline Compound 14a-1 (4.0 g, 13.3 mmol), potassium carbonate (3.7 g, 268 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (2.2 g, 10.7 mmol), and [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (972 mg, 1.34 mmol) were sequentially added to a three-neck flask containing 60 mL of 1,4-dioxane:water (3:1), and the mixture was reacted at 50 °C for 4 h under nitrogen protection. The reaction mixture was diluted with water and extracted with EA. The combined organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (PE:EA = 3:1) to give product 14a-2 (2.3 g, yellow solid, yield: 67.9%). 1 H NMR (400MHz, DMSO-d6): δ7.49(d,J=2.0Hz,1H),6.97(d,J=2.0Hz,1H),6.92(d,J =8.0Hz,1H),6.76-6.73(m,1H),6.26(d,J=2.0Hz,1H),5.21(s,2H),3.63(s,3H).

[0117] Step 2: Synthesis of 7-bromo-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 14a-2 (2.25 g, 8.9 mmol) and carbonyldiimidazole (4.35 g, 26.80 mmol) were added to a single-neck flask containing 30 mL of N-methylpyrrolidone, and the mixture was allowed to react overnight at 145 °C. The reaction mixture was diluted with water, and the solid precipitated. The solid was filtered and dried to give 14a-3 (1.6 g, brown-white solid, yield: 45.1%). LCMS (ESI): m / z 280 [M+H] + ;RT=1.410min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ11.50 (s, 1H), 8.14-8.10 (m, 2H), 7.64 (d, J = 2Hz, 1H), 7.46-7.44 (m, 1H), 4.34 (s, 3H).

[0118] Step 3: Synthesis of 7-acetyl-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 14a-3 (600 mg, 2.16 mmol), tributyltin (1.56 g, 4.32 mmol), and bis(triphenylphosphine)palladium(II) dichloride (227 mg, 0.32 mmol) were added to a single-neck flask containing 10 mL of dioxane, and the mixture was reacted at 95 °C for 16 hours. The reaction mixture was filtered, concentrated, and purified by column chromatography (DCM:MeOH = 20:1) to give 14a-4 (300 mg, yellow solid, 100% yield). LCMS (ESI): m / z 242.2 [M+H] + ;RT=1.057min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ11.60(s,1H),8.34(d,J=8.4Hz,1H),8.15(s,1H),8.04(d,J=1.6Hz,1H),7.86-7.84(m,1H),4.41(s,3H),2.65(s,3H).

[0119] Step 4: Synthesis of 7-(1-hydroxyethyl)-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one To a single-neck flask, 14a-4 (300 mg, 1.24 mmol), methanol (4 mL), and sodium borohydride (57 mg, 1.50 mmol) were added sequentially in an ice bath. The mixture was stirred at room temperature for 5 hours. The mixture was quenched with saturated ammonium chloride (1 mL), adjusted to neutral pH, diluted with water, and extracted with EA (10 mL × 5). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 14a-5 (50 mg, yellow solid, yield: 16.5%). LCMS (ESI): m / z 244.1 [M+H] + ;RT=0.860min(2.50min).

[0120] Step 5: Synthesis of 7-(1-bromoethyl)-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one 14a-5 (50 mg, 0.20 mmol), triphenylphosphine (157 mg, 0.60 mmol), and carbon tetrabromide (133 mg, 0.40 mmol) were added to a 100 mL single-neck flask containing 5 mL of DCM, and the mixture was allowed to react at room temperature for 2 hours. The reaction mixture was concentrated to give 14a (30 mg, yellow solid, yield: 47.7%). LCMS (ESI): m / z 306.0 [M+H] + ;RT=1.477min(2.50min).

[0121] Intermediate 15a: (1-methyl-4-oxo-4,5-dihydro-1H-pyrazolo[4,3-c]quinolin-7-yl)methyl methanesulfonate [ka]

[0122] Step 1: Synthesis of 7-(hydroxymethyl)-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one Compound 14a-3 (500 mg, 1.80 mmol), 1,4-dioxane (10 mL), and XPhos Pd G2 (142 mg, 0.18 mmol) were added to a dry 50 mL three-neck flask. Under nitrogen protection, (tributylstannyl)methanol (866 mg, 2.70 mmol) was added. The mixture was heated to 80 °C and reacted for 16 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was slurried in EA and filtered. The filter cake was collected to give 15a-1 (400 mg, black solid, yield: 97.05%). LCMS (ESI): m / z 230.1 [M+H] + ;RT=0.896min(2.50min).

[0123] Step 2: Synthesis of (1-methyl-4-oxo-4,5-dihydro-1H-pyrazolo[4,3-c]quinolin-7-yl)methyl methanesulfonate A dry one-neck flask was charged with 15a-2 (100 mg, 0.44 mmol), THF (5 mL), triethylamine (0.20 mL, 1.31 mmol), and methanesulfonyl chloride (0.06 mL, 0.87 mmol), sequentially. The reaction was allowed to proceed at room temperature for 16 h. The mixture was diluted with water (50 mL) and extracted with EA. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 15a (56 mg, yellow solid). LCMS (ESI): m / z 308.1 [M+H] + ;RT=1.083min(2.50min).

[0124] Intermediate 16a: 7-(1-bromoethyl)-6-chloro-1-methyl-1,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0125] The synthesis method is based on Intermediate 14a, except that Intermediate 11a-6 was used instead of Intermediate 14a-3. LCMS (ESI): m / z 326.0 [M+H] + ;RT=1.393min(2.50min).

[0126] Intermediate 17a: 6-chloro-7-(hydroxymethyl)-2-methyl-2,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0127] The synthetic method is based on Intermediate 11a, except that (1-methyl-1H-pyrazol-3-yl)boronic acid was used instead of intermediate 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole. LCMS (ESI): m / z 310.0 [M+H] + ;RT=1.327min(2.50min).

[0128] Intermediate 18a: 7-(1-bromoethyl)-6-chloro-2-methyl-2,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0129] For the synthesis method, see Intermediate 16a. LCMS (ESI): m / z 326.1 [M+H] + ;RT=0.599min(2.50min).

[0130] Intermediate 20a: 7-(1-bromoethyl)-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one [ka]

[0131] Step 1: Synthesis of 7-bromo-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one A dry one-neck flask was charged with 20a-1 (1.9 g, 6.4 mmol), absolute ethanol (30 mL), methylhydrazine sulfate (2.77 g, 19.2 mmol), and glacial acetic acid (0.5 mL). The reaction was allowed to proceed at 95 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, diluted with water, adjusted to pH 9, filtered, and the filter cake was dried under vacuum to give 20a-2 (1.3 g, white solid, yield: 72.9%). LCMS (ESI): m / z 280.0 [M+H] + ;RT=1.267min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ11.42(s,1H),8.57(s,1H),7.83(d,J=8.4Hz,1H),7.51(d,J=1.6Hz,1H),7.37-7.34(m,1H),4.13(s,3H).

[0132] Step 2: Synthesis of 7-acetyl-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one A dry three-neck flask was charged with 20a-2 (400 mg, 1.44 mmol), 1,4-dioxane (10 mL), and bis(triphenylphosphine)palladium(II) chloride (98 mg, 0.14 mmol). Under nitrogen protection, tributyl(1-ethoxyvinyl)tin (779 mg, 2.16 mmol) was added. The mixture was heated to 95 °C and reacted for 16 h. The temperature was lowered to 50 °C, and hydrochloric acid solution (1.0 M, 4 mL) was slowly added dropwise. The mixture was reacted for 1 h. Saturated potassium fluoride solution (10 mL) was added, and the mixture was stirred for 1 h and filtered. The filtrate was extracted with water and EA (20 mL × 3). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography (DCM:methanol=20:1) to give 20a-3 (200 mg, yellow solid, yield: 57.7%). LCMS (ESI): m / z 242.1 [M+H] + ;RT=0.877min(2.50min). 1H NMR (400MHz, DMSO-d6): δ11.50(s,1H),8.77(s,1H),8.00(d,J=8.0Hz,1H),7.93(d,J=1.6Hz,1H),7.80-7.78(m,1H),4.15(s,3H),2.61(s,3H).

[0133] Step 3: Synthesis of 7-(1-hydroxyethyl)-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one A dry 100 mL single-neck flask was charged with 20a-3 (140 mg, 0.58 mmol), methanol (4 mL), and sodium borohydride (21 mg, 0.58 mmol) in an ice bath. The mixture was stirred at room temperature for 3 hours. The mixture was diluted with water and extracted with DCM (15 mL x 4). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 20a-4 (110 mg, yellow solid). LCMS (ESI): m / z 244.1 [M+H] + ;RT=1.045min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ11.28(s,1H),8.59(s,1H),7.79(d,J=7.6Hz,1H),7.35(s,1H),7.14( d,J=8.0Hz,1H),5.26(d,J=4.0Hz,1H),4.77-4.74(m,1H),4.12(s,3H),1.34(d,J=6.4Hz,3H).

[0134] Step 4: Synthesis of 7-(1-bromoethyl)-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one A dry three-necked flask was charged with 20a-4 (100 mg, 0.41 mmol) and DCM (5 mL) in that order. Under nitrogen protection, PBr3 (333 mg, 1.23 mmol) was slowly added dropwise at 0 °C. The reaction was allowed to proceed at room temperature for 3 h. The reaction mixture was diluted with water (15 mL) and extracted with DCM (15 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 20a (100 mg, yellow solid). LCMS (ESI): m / z 306.1 [M+H]+ ;RT=1.054min(2.50min).

[0135] Intermediate 21a: 7-(bromomethyl)-2-methyl-2,5-dihydro-4H-pyrazole[3,4-c]quinolin-4-one [ka]

[0136] The synthesis method refers to intermediate 11a, except that 20a-2 was used instead of 11a-6. LCMS (ESI): m / z 292.0 [M+H] + ;RT=1.263min(2.50min).

[0137] Intermediate 22a: 7-(bromomethyl)-6-fluorothieno[2,3-c]quinolin-4(5H)-one [ka]

[0138] Step 1: Synthesis of methyl 3-bromothiophene-2-carboxylate 22a-1 (1 g, 4.83 mmol) was dissolved in methanol (20 mL), concentrated sulfuric acid (0.5 mL) was added dropwise, and the mixture was refluxed at 80° C. for 8 h. After cooling to room temperature, the mixture was concentrated, water (30 mL) was added, and the mixture was extracted three times with EA. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and dried in vacuo to give 22a-2 (950 mg, 4.30 mmol, white solid, yield: 89%).

[0139] Step 2: Synthesis of (2-(methoxycarbonyl)thiophen-3-yl)boronic acid A dry 50 mL three-neck flask was charged with 22a-2 (700 mg, 3.17 mmol), bis(pinacolato)diboron (1.61 g, 6.34 mmol), Pd(dppf)Cl2 (695 mg, 0.76 mmol), potassium acetate (932 mg, 9.51 mmol), and dioxane (20 mL) in that order, and the mixture was stirred at 100 °C for 8 h under nitrogen protection. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated and purified using a reverse-phase column (acetonitrile:water (1‰ NH4HCO3)) to give 22a-3 (40 mg, 0.215 mmol, white solid, 6.8% yield). LCMS (ESI): m / z 186.9 [M+H] + ;RT=2.61min(5.00min).

[0140] Step 3: Synthesis of methyl 3-(4-bromo-2-((t-butoxycarbonyl)amino)-3-fluorophenyl)thiophene-2-carboxylate 22a-3 (40 mg, 0.215 mmol), 11a-3 (98 mg, 0.236 mmol), Pd(ppf)Cl2 (16 mg, 0.0215 mmol), K2CO3 (89 mg, 0.645 mmol), dioxane (8 mL), and water (2 mL) were added to a 25 mL single-neck flask and stirred at 70 °C for 1 h under nitrogen protection. The reaction mixture was diluted with EA, washed twice with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified using a silica gel column (EA:PE = 1:5) to give 22a-4 (71 mg, 0.165 mmol, white solid, 76.7% yield). LCMS (ESI): m / z 331.9 [M+H-100] + ;RT=1.85min(3.00min).

[0141] Step 4: Synthesis of 7-bromo-6-fluorothieno[2,3-c]quinolin-4(5H)-one 22a-4 (71 mg, 0.165 mmol) was dissolved in methanol (4 mL) at room temperature, and HCl / dioxane (2 mL, 4 M) was added. The reaction mixture was stirred at 45 °C for 1 h. The reaction mixture was concentrated and dried under vacuum to give 22a-5 (49 mg, 0.148 mmol, white solid, yield: 90%). LCMS (ESI): m / z 299.9 [M+H]+ ;RT=1.46min(3.00min).

[0142] Step 5: Synthesis of 6-chloro-7-(hydroxymethyl)thieno[2,3-c]quinolin-4(5H)-one A 25 mL single-neck flask was charged with 22a-5 (49 mg, 0.148 mmol), (tributyltin)methanol (71 mg, 0.222 mmol), X-phos Pd G2 (11.6 mg, 0.0148 mmol), and dioxane (5 mL) in that order. The mixture was stirred at 100 °C for 4 h under nitrogen protection, concentrated, and purified using a silica gel column (EA:PE = 1:5) to give 22a-6 (31 mg, 0.124 mmol, white solid, 83.8% yield). LCMS (ESI): m / z 250.1 [M+H] + ;RT=0.95min(3.00min).

[0143] Step 6: 7-(Bromomethyl)-6-fluorothieno[2,3-c]quinolin-4(5H)-one 22a-6 (31 mg, 0.124 mmol), DCM (5 mL), and phosphorus tribromide (168 mg, 0.622 mmol) were added dropwise to a 25 mL single-neck flask at room temperature. The reaction mixture was stirred at room temperature for 2 hours and then concentrated to give crude product 22a (50 mg, 100%). LCMS (ESI): m / z 311.9 [M+H] + ;RT=1.41min (3.00min).

[0144] Intermediate 23a: 7-(bromomethyl)-6-fluorothieno[3,4-c]quinolin-4(5H)-one [ka]

[0145] The synthesis method is based on intermediate 22a, except that 4-bromothiophene-3-carboxylic acid was used instead of 22a-1. LCMS (ESI): m / z 311.9 [M+H] + ;RT=1.57min(3.00min).

[0146] Intermediate 24a: 6-(piperazin-1-yl)nicotinic acid hydrochloride [ka]

[0147] Step 1: Synthesis of t-butyl 4-(5-cyanopyridin-2-yl)piperazine-1-carboxylate A 100 mL three-neck flask was charged with 24a-1 (500 mg, 4.09 mmol), 1-t-butoxycarbonylpiperazine (915.24 mg, 4.91 mmol), and acetonitrile (10 mL), followed by dropwise addition of DIEA (1.36 mL, 8.19 mmol). The mixture was then reacted at 65 °C for 16 hours under nitrogen protection. The solvent was removed, and the residue was subjected to column chromatography (PE:EA = 5:1) to give 24a-2 (1.10 g, yellow solid, 93.22% yield). LCMS (ESI): m / z 289.2 [M+H] + ;RT=1.728min(2.50min).

[0148] Step 2: Synthesis of 6-(piperazin-1-yl)nicotinic acid hydrochloride 24a-2 (1.10 g, 3.81 mmol) and DCM (15 mL) were added sequentially to a dry 100 mL single-neck flask, and HCl-1,4-dioxane (1.5 mL) was added dropwise in an ice bath. The mixture was stirred at room temperature for 2 hours. The solvent was removed to give 24a (850 mg, yellow oil). LCMS (ESI): m / z 189.2 [M+H] + ;RT=0.352min. 1 H NMR (400MHz, DMSO-d6): δ9.54(s,2H),8.55(d,J=2.4Hz,1H),7.97-7.94(m,1H),7.03(d,J=9.2Hz,1H),3.93-3.91(m,4H),3.16(s,4H).

[0149] Intermediate 25a: 1-(2,4-difluorophenyl)piperazine [ka]

[0150] Step 1: Synthesis of t-butyl 4-(2,4-difluorophenyl)piperazine-1-carboxylate A dry 100 mL three-neck flask was charged with 25a-1 (2.00 g, 8.33 mmol), 1-t-butoxycarbonylpiperazine (2.33 g, 12.50 mmol), cesium carbonate (6.79 g, 20.83 mmol), 1,4-dioxane (50 mL), and Ruphos Pd G3 (698 mg, 0.83 mmol), and the mixture was heated to 100 °C under nitrogen protection for 16 h. After filtration, the solvent was removed, and the residue was subjected to column chromatography (PE:EA = 20:1) to give 25a-2 (720 mg, black solid, 25.00% yield). LCMS (ESI): m / z 243.1 [M+H-56]. + ;RT=1.624min(2.50min).

[0151] Step 2: Synthesis of 1-(2,4-difluorophenyl)piperazine 25a-2 (1.10 g, 3.81 mmol) and DCM (15 mL) were added sequentially to a dry 100 mL single-neck flask. HCl (1.5 mL) in 1,4-dioxane was added dropwise in an ice bath, and the mixture was stirred at room temperature for 2 h. The solvent was removed, and the mixture was extracted with water and EA. The aqueous phase was adjusted to weak alkalinity with saturated sodium bicarbonate and then extracted with EA. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give crude product 25a (180 mg, black oil). LCMS (ESI): m / z 199.1 [M+H] + ;RT=0.672min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ7.20-7.14 (m, 1H), 7.07-6.95 (m, 2H), 2.91-2.82 (m, 8H).

[0152] Intermediate 26a: 7-(bromomethyl)-2-methyl-2,5-dihydro-4H-pyrazolo[4,3-c]quinolin-4-one [ka]

[0153] The synthesis method refers to intermediate 11a, except that 5-bromo-2-iodoaniline was used instead of 11a-3. LCMS (ESI): m / z 292.0 [M+H] + ;RT=1.234min(2.50min).

[0154] Intermediate 27a: 7-(hydroxymethyl)-2-methyloxazolo[4,5-c]quinolin-4(5H)-one [ka]

[0155] Step 1: Synthesis of methyl (4-iodo-3-(2-methyloxazole-4-carboxamido)benzoate A dry 25 mL single-neck flask was charged with 27a-1 (340 mg, 2.68 mmol), methyl 3-amino-4-iodobenzoate (741 mg, 2.68 mmol), EA (20 mL), and T3P (5.12 g, 8.04 mmol, 50% EA solution) in that order, and the mixture was stirred at 80 °C for 16 h under nitrogen protection. After cooling to room temperature, water (20 mL) was added and the mixture was stirred for 2 min. The layers were separated, and the aqueous phase was extracted with EA. The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated, and purified on a silica gel column (EA:PE = 3:1) to give 27a-2 (420 mg, 1.09 mmol, pale yellow solid, 40.6% yield). LCMS (ESI): m / z 387.0 [M+H] + ;RT=1.74min (3.0min).

[0156] Step 2: Synthesis of methyl 3-(N-(t-butoxycarbonyl)-2-methyloxazole-4-carboxamido)-4-iodobenzoate To a dry 25 mL single-neck flask, 27a-2 (420 mg, 1.09 mmol), DCM (15 mL), di-t-butyl dicarbonate (356 mg, 1.63 mmol), and DMAP (199 mg, 1.63 mmol) were added sequentially and stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, stirred, and purified using a silica gel column (EA:PE = 1:5) to give product 27a-3 (350 mg, 0.72 mmol, white solid, 66% yield). LCMS (ESI): m / z 387.0 [M+H-100] + ;RT=1.80min(3.00min).

[0157] Step 3: Synthesis of methyl 2-methyl-4-oxo-4,5-dihydrooxazole[4,5-c]quinoline-7-carboxylate 27a-3 (320 mg, 0.658 mmol), Pd(OAc)2 (30 mg, 0.132 mmol), triphenylphosphine (34.6 mg, 0.132 mmol), and potassium carbonate (182 mg, 1.316 mmol) were added to DMF (5 mL). The reaction mixture was stirred under nitrogen protection at 100 °C for 1.5 h under microwave conditions. The reaction mixture was filtered and washed with EA. The filtrate was concentrated and then purified by reverse-phase column (acetonitrile:water (1‰HCOOH)) and lyophilized to give product 27a-4 (140 mg, 0.534 mmol, off-white solid, yield: 82.5%).

[0158] LCMS(ESI):m / z259.1[M+H] + ;RT=1.28min(3.00min).

[0159] Step 4: Synthesis of 7-(hydroxymethyl)-2-methyloxazolo[4,5-c]quinolin-4(5H)-one 27a-4 (140 mg, 0.534 mmol) was dissolved in THF (10 mL), and LiAlH (1.08 mL, 1M / THF) was added dropwise in an ice bath. The mixture was stirred continuously for 2 hours. The reaction mixture was quenched with methanol, the pH adjusted to weakly acidic with trifluoroacetic acid, and concentrated under reduced pressure. The residue was dissolved in dimethyl sulfoxide (5 mL) and purified using a reverse-phase column (acetonitrile:water (1‰NHHCO)) to give 27a-5 (75 mg, 0.326 mmol, off-white solid, 60% yield). LCMS (ESI): m / z 231.1 [M+H] + ;RT=1.07min(3.00min).

[0160] Step 5: Synthesis of 7-(bromomethyl)-2-methyloxazolo[4,5-c]quinolin-4(5H)-one 27a-5 (40 mg, 0.174 mmol) and DCM (5 mL) were added sequentially to a dry 25 mL single-neck flask at room temperature, and phosphorus tribromide (235 mg, 0.869 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give an oil (60 mg, crude product). LCMS (ESI): m / z 292.9 [M+H] + ;RT=1.39min(3.00min).

[0161] Intermediate 28a: 7-(chloromethyl)-2-methylthiazolo[4,5-c]quinolin-4(5H)-one [ka]

[0162] Step 1: Synthesis of ethyl 5-bromo-2-methylthiazole-4-carboxylate 28a-1 (2.0 g, 11.7 mmol), N-bromosuccinimide (4.16 g, 23.4 mmol), and anhydrous acetonitrile (20 mL) were added to a 50 mL single-neck flask, heated to 90 °C under argon protection, and stirred for 16 h. The mixture was cooled to room temperature, concentrated under reduced pressure, and passed through a normal phase column (PE:EA = 5:1) to give 28a-2 (1.3 g, yellow solid, 45% yield). LCMS (ESI): m / z 250.1 [M+H]+ ;RT=1.30min (3.0min).

[0163] Step 2: Synthesis of methyl 2-methyl-4-oxo-4,5-dihydrothiazole[4,5-c]quinoline-7-carboxylate 28a-2 (100 mg, 0.4 mmol), 2-amino-4-methoxycarbonylphenylboronic acid pinacol ester (166 mg, 0.6 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (29 mg, 0.04 mmol), potassium carbonate (166 mg, 1.2 mmol), dioxane (5 mL), and water (1 mL) were added to a single-neck flask. The mixture was heated to 80 °C under argon protection and stirred for 16 h. The mixture was cooled to room temperature, water was added, and the mixture was extracted with EA. The organic phase was dried over anhydrous sodium sulfate, and the residue was passed through a normal phase column (DCM:methanol = 10:1) to give 28a-3 (50 mg, white solid, yield: 45%). LCMS (ESI): m / z 275.1 [M+H] + ;RT=1.03min (3.0min).

[0164] Step 3: Synthesis of 7-(hydroxymethyl)-2-methylthiazolo[4,5-c]quinolin-4(5H)-one 28a-3 (50 mg, 0.18 mmol) and anhydrous THF (5 mL) were added to a 50 mL one-neck flask, cooled to 0 °C, and 1 M lithium aluminum tetrahydride solution (0.5 mL, 0.5 mmol) was slowly added dropwise. The mixture was stirred for 2 h, quenched with methanol, and 2 mL of trifluoroacetic acid was added dropwise. The mixture was stirred for 10 min and concentrated. The residue was passed through a reverse-phase column (1%-30% acetonitrile / 0.1% formic acid aqueous solution) to give 28a-4 (30 mg, white solid, 45% yield). LCMS (ESI): m / z 247.1 [M+H] + ;RT=0.54min (3.0min).

[0165] Step 4: Synthesis of 7-(chloromethyl)-2-methylthiazolo[4,5-c]quinolin-4(5H)-one 28a-4 (30 mg, 0.12 mmol) and anhydrous DCM (5 mL) were added to a 50 mL one-neck flask, cooled to 0 °C, and thionyl chloride (1 mL) was slowly added dropwise. The mixture was stirred for 2 hours and concentrated under reduced pressure to give crude product 28a (30 mg, white solid, 100% yield). LCMS (ESI): m / z 265.1 [M+H] + ;RT=1.41min (3.0min).

[0166] Intermediate 31a: 6-chloro-7-(bromomethyl)-[1,2,3]triazolo[1,5-a]quinoxalin-4(5H)-one [ka]

[0167] Step 1: Synthesis of 6-bromo-5-chloro-3-oxo-3,4-dihydroquinoxaline-2-formaldehyde 31a-1 (500 mg, 1.9 mmol), tin dioxide (328 mg, 2.9 mmol), and dioxane (10 mL) were added sequentially to a dry 50 mL single-neck flask at room temperature. The mixture was heated to 100 °C and stirred overnight. The reaction mixture was filtered, and the filtrate was concentrated to give 31a-2 (600 mg, oil). LCMS (ESI): m / z 271.2 [M+H] + ;RT=1.11min (3.00min).

[0168] Step 2: Synthesis of 7-bromo-6-chloro-[1,2,3]triazolo[1,5-a]quinoxalin-4(5H)-one 31a-2 (700 mg, 2.6 mmol), 4-methylbenzenesulfonyl hydrazide (579 mg, 3.1 mmol), and methanol (10 mL) were added to a dry one-neck flask and stirred at room temperature for 4 hours. The filtrate was concentrated, and the residue was purified by reverse-phase preparative column chromatography (1% to 50% acetonitrile) to give product 31a-3 (600 mg, red solid, 71% yield). LCMS (ESI): m / z 282.8 [M+H] + ;RT=1.617min(3.00min).

[0169] Step 3: Synthesis of 6-chloro-7-(hydroxymethyl)-[1,2,3]triazolo[1,5-a]quinoxalin-4(5H)-one A dry 50 mL single-neck flask was charged with 31a-3 (200 mg, 0.71 mmol), (tributyltin)methanol (273 mg, 0.85 mmol), x-phos Pd Glu (56 mg, 0.071 mmol), and dioxane (10 mL) at room temperature. The mixture was heated overnight at 80 °C under nitrogen protection. The reaction mixture was concentrated and purified using a silica gel column (DCM:methanol = 30:1) to give 31a-4 (100 mg, white solid, 60% yield). LCMS (ESI): m / z 234.9 [M+H] + ;RT=1.317min(3.00min).

[0170] Step 4: Synthesis of 6-chloro-7-(bromomethyl)-[1,2,3]triazolo[1,5-a]quinoxalin-4(5H)-one A dry 50 mL single-neck flask was charged with 31a-4 (20 mg, 0.085 mol) and dioxane (3 mL) at room temperature, followed by dropwise addition of phosphorus tribromide (69 mg, 0.25 mmol) in an ice bath. The mixture was stirred at room temperature for 2 hours, and the filtrate was concentrated to give the crude product (40 mg, oil). LCMS (ESI): m / z 296.9 [M+H] + ;RT=1.45min(3.00min).

[0171] Intermediate 32a: 7-(1-bromoethyl)-6-chloro-[1,2,3]triazolo[1,5-a]quinoxalin-4(5H)-one [ka]

[0172] The synthesis method was the same as that of 31a, except that the reaction conditions for the Stille coupling reaction were tributyl(1-ethoxyvinyl)tin, PdCl2(PPh3)2. LCMS (ESI): m / z 310.9 [M+H] + ;RT=1.55min(3.00min).

[0173] Intermediate 33a: 8-(chloromethyl)imidazo[1,2-c]quinazolin-5(6H)-one [ka]

[0174] Step 1: Synthesis of t-butyl 2-bromo-1H-imidazole-1-carboxylate 33a-1 (200 mg, 1.36 mmol), di-t-butyl dicarbonate (445 mg, 2.04 mmol), triethylamine (412 mg, 4.08 mmol), and DMF (5 mL) were added to a 50 mL one-neck flask and stirred at room temperature under argon protection for 2 h. The reaction mixture was extracted with water and EA. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified using a normal phase column (PE:EA = 5:1) to give 33a-2 (250 mg, white solid, yield: 74%). LCMS (ESI): m / z 146.9 [M+H-100] + ;RT=1.58min (3.0min).

[0175] Step 2: Synthesis of methyl 5-carbonyl-5,6-dihydroimidazo[1,2-c]quinazoline-8-carboxylate 33a-2 (100 mg, 0.4 mmol), methyl 3-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (166 mg, 0.6 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (29 mg, 0.04 mmol), potassium carbonate (166 mg, 1.2 mmol), dioxane (5 mL), and water (1 mL) were added to a 50 mL one-neck flask and stirred at 80 °C for 16 h under argon protection. The mixture was cooled to room temperature, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, and the residue was purified by normal phase column chromatography (DCM:methanol = 10:1) to give 33a-3 (50 mg, white solid, 45% yield). LCMS (ESI): m / z 244.1 [M+H] + ;RT=1.22min (3.0min).

[0176] Step 3: Synthesis of 8-(hydroxymethyl)imidazo[1,2-c]quinazolin-5(6H)-one 33a-3 (50 mg, 0.20 mmol) and anhydrous THF (5 mL) were added to a 50 mL one-neck flask, cooled to 0 °C, and 1 M lithium aluminum tetrahydride solution (0.5 mL, 0.5 mmol) was slowly added dropwise. The mixture was stirred for 2 h, quenched with methanol, and 2 mL of trifluoroacetic acid was added dropwise. The mixture was stirred for 10 min and concentrated. The residue was purified using a reverse-phase column (1%-30% acetonitrile / 0.1% aqueous ammonium bicarbonate) to give 33a-4 (20 mg, white solid, 47% yield). LCMS (ESI): m / z 216.1 [M+H] + ;RT=0.99min(3.0min).

[0177] Step 4: Synthesis of 8-(chloromethyl)imidazo[1,2-c]quinazolin-5(6H)-one 33a-4 (20 mg, 0.09 mmol) and DCM (5 mL) were added to a single-neck flask, cooled to 0 °C, and thionyl chloride (1 mL) was slowly added dropwise. The mixture was stirred for 2 hours and concentrated under reduced pressure to give crude product 33a (20 mg, white solid). LCMS (ESI): m / z 234.1 [M+H] + ;RT=1.28min (3.0min).

[0178] Intermediate 34a: Synthesis of N-ethyl-6-chloro-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0179] The synthesis method is based on the synthesis of intermediate 2a, except that ethylamine was used instead of methylamine hydrochloride in the condensation step. LCMS (ESI): m / z 253.2 [M+H] + ;RT=0.527min (2.5min).

[0180] Intermediate 35a: (R)-6-chloro-5-(piperazin-1-yl)-N-(tetrahydrofuran-3-yl)pyridinecarboxamide hydrochloride [ka]

[0181] The synthesis method refers to the synthesis of intermediate 2a, except that (R)-tetrahydrofuran-3-amine was used instead of methylamine hydrochloride in the condensation step. LCMS (ESI): m / z 395.2 [M+H] + ;RT=1.582min(2.50min).

[0182] Intermediate 36a: 6-chloro-N-methyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0183] Step 1: Synthesis of 5-bromo-6-chloro-N-methylpyridinecarboxamide 36a-1 (1.00 g, 3.99 mmol) and a solution of methylamine in ethanol (33% wt, 10 mL) were added sequentially to a dry 100 mL single-neck flask. The mixture was allowed to react at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to give 36a-2 (0.94 g, yellow oil, yield: 94.37%). LCMS (ESI): m / z 250.9 [M+H] + ;RT=1.448min(2.50min).

[0184] Step 2: Synthesis of t-butyl 4-(2-chloro-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate A dry 50 mL single-neck flask was charged with 36a-2 (620 mg, 2.49 mmol), toluene (15 mL), piperazine-1-t-butylcarboxylate (370 mg, 1.99 mmol), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (155 mg, 0.25 mmol), cesium carbonate (2020 mg, 6.21 mmol), and palladium acetate (56 mg, 0.25 mmol), sequentially. The mixture was heated to 100 °C under nitrogen protection and reacted for 16 h. The reaction mixture was filtered, and the filtrate was collected and concentrated under reduced pressure. Purification by silica gel column chromatography (PE:EA = 1:1) afforded 36a-3 (180 mg, yellow oil, 20.41% yield). LCMS (ESI): m / z 355.1 [M+H] + ;RT=1.679min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ8.46(d,J=4.8Hz,1H),7.95(d,J=8.4Hz,1H),7.68(d,J =8.4Hz,1H),3.50(s,4H),3.06-3.04(m,4H),2.80(d,J=4.8Hz,3H),1.43(s,9H).

[0185] Step 3: Synthesis of 6-chloro-N-methyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride A dried flask was charged with 36a-3 (180 mg, 0.51 mmol), DCM (2 mL), and HCl-dioxane (4.0 M, 2 mL). The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give 36a (140 mg, yellow solid, yield: 94.59%). LCMS (ESI): m / z 255.1 [M+H] + ;RT=0.336min&0.461min(2.50min). 1 H NMR(400MHz,DMSO-d6):δ9.40(s,2H),8.50(d,J=4.4Hz,1H),7.97(d,J=8.4Hz,1 H),7.76(d,J=8.4Hz,1H),3.34-3.32(m,4H),3.25(m,4H),2.80(d,J=4.8Hz,3H).

[0186] Intermediate 37a: N,6-dimethyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0187] Step 1: Synthesis of 5-bromo-6-methylpicolinic acid A dried flask was charged with 37a-1 (500 mg, 2.54 mmol), methanol (6 mL), water (3 mL), and sodium hydroxide (507 mg, 12.69 mmol), in that order. The mixture was allowed to react at 70°C for 1 hour. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (10 mL), adjusted to pH 4 with 3M dilute hydrochloric acid, and filtered to obtain 37a-2 (300 mg, white solid, 54.72% yield). LCMS (ESI): m / z 218.0 [M+H] + ;RT=1.208min(2.50min). 1 H NMR (400MHz, DMSO-d6): δ8.12 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 2.67 (s, 3H).

[0188] Step 2: Synthesis of 5-bromo-N,6-dimethylpyridinecarboxamide A dry flask was charged with 37a-2 (300 mg, 1.39 mmol), DMF (3 mL), DIEA (0.92 mL, 5.55 mmol), HATU (792 mg, 2.08 mmol), and a solution of methylamine in THF (2.0 M, 1.39 mL, 2.78 mmol). The reaction was allowed to proceed at room temperature for 1 h. The mixture was diluted with water (30 mL) and extracted with EA (10 mL × 2). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative plate chromatography (PE:EA = 3:2) to give 37a-3 (215 mg, yellow solid, yield: 67.59%). LCMS (ESI): m / z 231.0 [M+H] + ;RT=1.427min(2.50min). 1H NMR (400MHz, DMSO-d6): δ8.67(d,J=4.0Hz,1H),8.18(d,J=8.4Hz,1H),7.75(d,J=8.4Hz,1H),2.82(d,J=4.8Hz,3H),2.65(s,3H).

[0189] Step 3: Synthesis of t-butyl 4-(2-methyl-6-(methylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate A dried flask was charged with 37a-3 (200 mg, 0.87 mmol), toluene (8 mL), piperazine-1-t-butylcarboxylate (179 mg, 0.96 mmol), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (54 mg, 0.09 mmol), cesium carbonate (711 mg, 2.18 mmol), and palladium acetate (20 mg, 0.09 mmol). The mixture was heated to 100 °C under nitrogen protection for 16 h. The reaction mixture was filtered, and the filtrate was collected and concentrated under reduced pressure. The residue was purified by preparative plate (PE:EA = 1:1) to give 37a-4 (160 mg, yellow solid, yield: 54.80%). LCMS (ESI): m / z 335.1 [M+H] + ;RT=1.623min(2.50min). 1 H NMR(400MHz,DMSO-d6):δ8.44(d,J=4.8Hz,1H),7.80(d,J=8.4Hz,1H),7.49(d,J=8.4Hz ,1H),3.49(s,4H),2.89-2.87(m,4H),2.81(d,J=4.8Hz,3H),2.51(s,3H),1.43(s,9H).

[0190] Step 4: Synthesis of N,6-dimethyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride A dried flask was charged with 37a-4 (160 mg, 0.48 mmol), DCM (2 mL), and HCl-dioxane (4.0 M, 2 mL). The reaction was allowed to proceed at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to give 37a (129 mg, yellow solid, yield: 99.58%). LCMS (ESI): m / z 235.2 [M+H] +;RT=0.340min&0.450min(2.50min). 1 H NMR(400MHz,DMSO-d6):δ9.44(s,2H),8.56(d,J=4.8Hz,1H),7.88(d,J=8.0Hz,1 H),7.61(d,J=8.4Hz,1H),3.25-3.16(m,8H),2.82(d,J=4.4Hz,3H),2.54(s,3H).

[0191] Intermediate 38a: Synthesis of N-cyclopropyl-6-chloro-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0192] The synthesis method is based on the synthesis of intermediate 2a, except that cyclopropylamine was used instead of methylamine hydrochloride in the condensation step. LCMS (ESI): m / z 264.2 [M+H] + ;RT=0.958min(2.5min).

[0193] Intermediate 40a: 8-(bromomethyl)-2-methylimidazo[1,2-c]quinazolin-5(6H)-one [ka]

[0194] The synthesis method is the same as that of Intermediate 33a, except that 2-bromo-4-methyl-1H-imidazole was used as the starting material in Step 1 and phosphorus tribromide was used as the reaction reagent in Step 4. LCMS (ESI): m / z 292.0 [M+H] + ;RT=1.25min(3.00min).

[0195] Intermediate 41a: Synthesis of 8-(bromomethyl)-7-chloro-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0196] Step 1: Synthesis of 3-bromo-6-(4,5-dihydro-1H-imidazol-2-yl)-2-fluoroaniline Compound 41a-1 (500 mg, 2.33 mmol) was dissolved in methanol (20 mL), and ethylenediamine dihydrochloride (3.1 g, 23.3 mmol) and sodium carbonate (3.7 g, 35 mmol) were added sequentially. The mixture was heated to reflux for 72 hours. The mixture was concentrated, water (20 mL) was added, and the mixture was filtered. The solid was dissolved in DMSO (5 mL) and purified by reverse-phase column chromatography (acetonitrile:water) to give 41a-2 (130 mg, 0.504 mmol, white solid, 21.6% yield). LCMS (ESI): m / z 260.0 [M+H] + ;RT=0.75min (3.00min).

[0197] Step 2: Synthesis of 8-bromo-7-chloro-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 41a-2 (50 mg, 0.194 mmol) was dissolved in DMF (5 mL), and N,N'-carbonyldiimidazole (94 mg, 0.582 mmol) and DMAP (24 mg, 0.194 mmol) were added sequentially. The mixture was stirred at 100 °C for 3 h. After the reaction mixture was cooled to room temperature, the solid was collected by filtration, washed with EA, and dried under vacuum to give 41a-3 (41 mg, 0.144 mmol, white solid, 74.2% yield). LCMS (ESI): m / z 286.0 [M+H] + ;RT=1.22min (3.00min).

[0198] Step 3: Synthesis of 7-chloro-8-(hydroxymethyl)-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 41a-3 (41 mg, 0.144 mmol), (tributyltin)methanol (69 mg, 0.216 mmol), and x-phos Pd Glutamate (11.3 mg, 0.0144 mmol) were added sequentially to dioxane (5 mL). The mixture was stirred at 100 °C for 6 h under nitrogen protection, concentrated, and purified by silica gel column chromatography (methanol:DCM = 1:10) to give 41a-4 (29 mg, 0.123 mmol, white solid, yield: 85.7%). LCMS (ESI): m / z 236.1 [M+H]; RT = 0.91 min (3.00 min).

[0199] Step 4: Synthesis of 8-(bromomethyl)-7-chloro-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 41a-4 (29 mg, 0.123 mmol) was dissolved in DCM (8 mL), and phosphorus tribromide (0.5 mL) was added dropwise at room temperature. The mixture was stirred for 2 hours. The reaction mixture was concentrated to remove excess phosphorus tribromide and dried to give crude product 41a (35 mg). LCMS (ESI): m / z 298.2 [M+H] + ;RT=1.23min (3.00min).

[0200] Intermediate 42a: Synthesis of 8-(bromomethyl)-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0201] The synthesis method was the same as for 41a, except that 2-amino-4-bromobenzonitrile was used as the starting material. LCMS (ESI): m / z 280.0 [M+H] + ;RT=0.580min(2.50min).

[0202] Intermediate 43a: Synthesis of 8-(bromomethyl)-7-methyl-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0203] The synthesis method was the same as for 41a, except that 2-amino-4-bromo-3-methylbenzonitrile was used as the starting material. LCMS (ESI): m / z 294.1 [M+H] + ;RT=1.30min(3.00min).

[0204] Intermediate 44a: Synthesis of N-cyclopropyl-6-methyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0205] The synthesis method was the same as for 37a except that cyclopropylamine was used as the starting material. LCMS (ESI): m / z 261.1 [M+H] + ;RT=1.05min (3.0min).

[0206] Synthesis of intermediates 45a: 8-(bromomethyl)-7-chloro-dihydroimidazo[1,2-c]quinazolin-5(3H)-one, and 46a: 8-(bromomethyl)-9-chloro-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0207] Step 1: Synthesis of 5-bromo-2-(4,5-dihydro-1H-imidazol-2-yl)aniline 45a-1 (5 g, 25.5 mmol), diphosphorus pentasulfide (487 mg, 2.55 mmol), and ethylenediamine (10 mL) were added to a dry 100 mL single-neck flask at room temperature, and the mixture was allowed to react at 100 °C for 4 hours. Water (100 mL) was added to the reaction mixture, which was then suction filtered. The filter cake was washed with water and dried to give 45a-2 (1.5 g, pale yellow solid, 82% yield). LCMS (ESI): m / z 240.1 [M+H] + ;RT=0.98min(3.00min).

[0208] Step 2: Synthesis of 3-bromo-2-chloro-6-(4,5-dihydro-1H-imidazol-2-yl)aniline A dry 50 mL single-neck flask was charged with 45a-2 (500 mg, 2.09 mmol), N-chlorosuccinimide (335 mg, 2.5 mmol), p-toluenesulfonic acid (40 mg, 0.21 mmol), and DMF (10 mL) in that order, and the mixture was allowed to react at 90 °C for 6 h. Water (30 mL) was added to the reaction mixture, which was then extracted with EA (40 mL). The organic phase was washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated, and the residue was purified using a silica gel column (PE:EA = 3:1) to give a mixture of 45a-3 and 45a-4 (100 mg, yellow solid, 17% yield). LCMS (ESI): m / z 274.0 [M+H] + ;RT=1.19&1.21min(3.00min).

[0209] Step 3: Synthesis of 8-bromo-7-chloro-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one A mixture of 45a-3 and 45a-4 (100 mg, 0.37 mmol), N,N'-carbonyldiimidazole (118 mg, 0.73 mmol), DMAP (45 mg, 0.37 mmol), and DMF (3 mL) were added to a single-neck flask, heated to 100 °C, and reacted for 2 hours. Water (10 mL) was added to the reaction mixture, which was then suction filtered. The filter cake was washed with water and dried to give a mixture of 45a-5 and 45a-6 (90 mg, yellow solid, 82% yield). LCMS (ESI): m / z 299.9 [M+H] + ;RT=1.29&1.32min(3.00min).

[0210] Step 4: Synthesis of 7-chloro-8-(hydroxymethyl)-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one A mixture of 45a-5 and 45a-6 (90 mg, 0.3 mmol), (tributyltin)methanol (192 mg, 0.6 mmol), x-Phos Pd G2 (23 mg, 0.03 mmol), and dioxane (5 mL) were added to a dried flask and reacted at 100 °C for 6 hours under nitrogen protection. The mixture was concentrated, and the residue was purified using a reverse-phase preparative column (1% to 50% acetonitrile) to give 45a-7 (15 mg, yellow solid) and 45a-8 (25 mg, yellow solid).

[0211] 45a-7: LCMS (ESI): m / z 252.1 [M+H] + ;RT=0.95min(3.00min). 1 H NMR (600MHz, DMSO-d6): δ7.80 (d, J = 8.0 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 3.96 (m, 2H), 3.87 (m, 2H).

[0212] Step 5: Synthesis of 8-(bromomethyl)-7-chloro-dihydroimidazo[1,2-c]quinazolin-5(3H)-one A single-necked flask was charged with 45a-7 (15 mg, 0.06 mmol) and dioxane (3 mL), and phosphorus tribromide (48 mg, 0.18 mmol) was added dropwise in an ice bath. The mixture was stirred at room temperature for 2 hours and concentrated to give crude product 45a (20 mg, yellow solid). LCMS (ESI): m / z 314.0 & 316.0 [M+H] + ;RT=1.28min(3.00min).

[0213] Step 6: Synthesis of 8-(bromomethyl)-9-chloro-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one

[0214] A single-necked flask was charged with 45a-8 (25 mg, 0.1 mmol) and dioxane (3 mL), and phosphorus tribromide (81 mg, 0.3 mmol) was added dropwise in an ice bath. The mixture was stirred at room temperature for 2 hours and concentrated to give 46a (30 mg, yellow solid). LCMS (ESI): m / z 314.0 & 316.0 [M+H] + ;RT=1.32min(3.00min).

[0215] Intermediate 47a: Synthesis of 8-(bromomethyl)-7-chloro-2,2-dimethyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0216] Step 1: Synthesis of 2-((7-bromo-2-chloro-8-fluoroquinazolin-4-yl)amino)-2-methylprop-1-ol 47a-1 (300 mg, 1.01 mmol) was dissolved in THF (5 mL), and 47a-2 (108 mg, 1.21 mmol) and triethylamine (306 mg, 3.03 mmol) were added sequentially. The mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water (15 mL) and extracted three times with EA (20 mL). The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give 47a-3 (320 mg, 0.92 mmol, white solid, 91% yield). LCMS (ESI): m / z 349.9 [M+H] + ;RT=1.62min (3.0min).

[0217] Step 2: Synthesis of 8-bromo-5-chloro-7-chloro-2,2-dimethyl-2,3-dihydroimidazo[1,2-c]quinazoline 47a-3 (320 mg, 0.92 mmol) was dissolved in toluene (10 mL), and phosphorus oxychloride (1 mL) was added dropwise in an ice bath. The mixture was allowed to react at room temperature for 3 hours. The pH was adjusted to 7-8 with aqueous sodium carbonate, extracted with EA, dried over anhydrous sodium sulfate, concentrated, and purified on a silica gel column (PE:EA = 1:1) to give 47a-4 (215 mg, 0.651 mmol, white solid, yield, 70.8%). LCMS (ESI): m / z 331.9 [M+H] + ;RT=1.62min (3.0min).

[0218] Step 3: Synthesis of 8-bromo-7-chloro-2,2-dimethyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 47a-4 (215 mg, 0.651 mmol) was added to acetic acid (5 mL) and heated to 110 °C for 2 h. The reaction mixture was concentrated to remove acetic acid, and water (15 mL) was added to the residue. The pH was adjusted to 7-8 with saturated sodium bicarbonate solution, and the mixture was extracted three times with EA (20 mL). The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give 47a-5 (180 mg, 0.577 mmol, white solid, yield: 93.8%). LCMS (ESI): m / z 314.0 [M+H] + ;RT=1.32min (3.0min).

[0219] Step 4: 7-chloro-8-(hydroxymethyl)-2,2-dimethyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 47a-5 (80 mg, 0.256 mmol), (tributyltin)methanol (165 mg, 0.513 mmol), and X-Phos Pd G2 (20 mg, 0.0256 mmol) were added to anhydrous dioxane (5 mL), heated to 110 °C under argon protection, stirred for 2 hours, concentrated, and the residue was purified by column chromatography (DCM:methanol = 10:1) to give 47a-6 (48 mg, 0.154 mmol, white solid, yield: 60%). LCMS (ESI): m / z 264.2 [M+H] + ;RT=1.02min (3.0min).

[0220] Step 5: Synthesis of 8-(bromomethyl)-7-chloro-2,2-dimethyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one 47a-6 (48 mg, 0.154 mmol) was dissolved in DCM (10 mL), and phosphorus tribromide (0.5 mL) was added dropwise at room temperature. After the addition was complete, the mixture was stirred for an additional 2 hours. The reaction mixture was concentrated to remove the solvent and excess phosphorus tribromide, and the crude product 47a (60 mg) was dried under vacuum. LCMS (ESI): m / z 326.2 [M+H] + ;RT=1.38min (3.0min).

[0221] Intermediates 48a to 56a, 59a to 60a: The synthesis method was the same as that for 41a or 47a, except that the starting intermediates listed in the table below were used instead of ethylenediamine or 47a-2.

[0222] [Table 3] TIFF2025538192000063.tif94170

[0223] Intermediate 57a: 9-(chloromethyl)-3,3,8-trichloro-2,3,4,7-tetrahydro-6-pyrimido[1,2-c]quinazolin-6-one [ka]

[0224] Compound 57a-1 (20 mg, 0.07 mmol) was synthesized in the same manner as in 47a-6, except that 3-amino-2,2-difluoroprop-1-ol was used instead of 47a-2. Anhydrous DCM (10 mL) was added to a 50 mL one-neck flask, cooled to 0 °C, and thionyl chloride (1 mL) was slowly added dropwise. The mixture was stirred for 2 hours and concentrated under reduced pressure to give crude compound 57a (25 mg, white solid, 100% yield). LCMS (ESI): m / z 304.1 [M+H]. + ;RT=1.50min (3.0min).

[0225] Intermediate 58a: 9-(chloromethyl)-8-chloro-4-methyl-2,3,4,7-tetrahydro-6-pyrimido[1,2-c]quinazolin-6-one [ka]

[0226] The synthesis method was the same as for 57a, except that 4-amino-2-butanol was used instead of 47a-2. LCMS (ESI): m / z 282.1 [M+H] + ;RT=1.28min (3.0min).

[0227] Intermediate 61a: N-ethyl-6-methyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0228] The synthesis method was the same as for 37a, except that ethyl was used instead of methyl. LCMS (ESI): m / z 249.3 [M+H] + ;RT=1.0min (3.00min).

[0229] Intermediate 62a: 6-chloro-N-ethyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0230] The synthesis was the same as for 36a, except that ethyl was used instead of methyl. LCMS (ESI): m / z 269.3 [M+H] + ;RT=1.2min (3.00min).

[0231] Intermediate 63a: 6-chloro-N-cyclopropyl-5-(piperazin-1-yl)pyridinecarboxamide hydrochloride [ka]

[0232] The synthesis was the same as for 36a, except that cyclopropyl was used instead of methyl. LCMS (ESI): m / z 281.3 [M+H] + ;RT=1.3min (3.00min).

[0233] Intermediate 64a: 8'-(1-chloroethyl)-7'-chloro-3'-H-spiro[cyclopropane-1,2'-imidazo[1,2-c]quinazoline]-5'(6'H)-one [ka]

[0234] Step 1: 8'-Acetyl-7'-chloro-3'-H-spiro[cyclopropane-1,2'-imidazo[1,2-c]quinazoline]-5'(6'H)-one Compound 64a-1 (50 mg, 0.16 mmol), 64a-2 (87 mg, 0.24 mmol), and bis(triphenylphosphine)palladium(II) dichloride (11 mg, 0.016 mmol) were added to 5 mL of anhydrous dioxane and stirred at 100 °C for 6 h. After concentration, the residue was added with 5 N dilute hydrochloric acid (10 mL) and stirred at room temperature for 1 h. The mixture was then neutralized to alkalinity with saturated aqueous sodium bicarbonate and extracted with EA. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated, and purified by normal phase column chromatography (PE:EA = 3:1) to give compound 64a-3 (20 mg, white solid, yield: 46%). LCMS (ESI): m / z 274.1 [M+H] + ;RT=1.21min (3.0min).

[0235] Step 2: 7'-Chloro-8'-(1-hydroxyethyl)-3'-H-spiro[cyclopropane-1,2'-imidazo[1,2-c]quinazoline]-5'(6'H)-one 64a-3 (50 mg, 0.18 mmol) was dissolved in anhydrous methanol (10 mL), sodium borohydride (69 mg, 1.8 mmol) was added, and the mixture was stirred at 0 °C for 2 h. The mixture was concentrated under reduced pressure and extracted with ethyl acetate (30 mL × 3) and water (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to give compound 64a-4 (30 mg, white solid, yield: 60%). LCMS (ESI): m / z 276.2 [M+H] + ;RT=1.09min(3.0min).

[0236] Step 3: 8'-(1-chloroethyl)-7'-chloro-3'-H-spiro[cyclopropane-1,2'-imidazo[1,2-c]quinazoline]-5'(6'H)-one Compound 64a-4 (20 mg, 0.073 mmol) was dissolved in anhydrous dichloromethane (10 mL), cooled to 0 °C, and thionyl chloride (1 mL) was slowly added dropwise. The mixture was stirred for 2 hours and concentrated under reduced pressure to give crude product 64a (20 mg, white solid, yield: 100%). LCMS (ESI): m / z 294.1 [M+H] + ;RT=1.38min (3.0min).

[0237] Intermediate 65a: (2R)-8-(1-bromoethyl)-7-chloro-2-methyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one [ka]

[0238] The synthesis method was the same as for 64a, except that (R)-8-bromo-7-chloro-2-methyl-2,6-dihydroimidazo[1,2-c]quinazolin-5(3H)-one was used instead of 64a-1. LCMS (ESI): m / z 326.0 [M+H] + ;RT=1.37min (3.0min).

[0239] Intermediate 66a: Methyl (R)-(2-methyl-5-oxo-2,3,5,6-tetrahydroimidazo[1,2-c]quinazolin-8-yl)methanesulfonate [ka]

[0240] To a 50 mL single-neck flask, 66a-1 (100 mg, 0.43 mmol), THF (10 mL), triethylamine (0.18 mL, 1.29 mmol), and methanesulfonyl chloride (0.05 mL, 0.65 mmol) were added sequentially at 0 °C. The reaction was allowed to proceed at room temperature for 2 h. The reaction mixture was concentrated to give 66a (120 mg, crude product, yellow solid). LCMS (ESI): m / z 310.0 [M+H] + ;RT=0.807min(2.50min).

[0241] Intermediate 67a: 9-(Bromomethyl)-2,3,4,7-tetrahydro-6-pyrimido[1,2-c]quinazolin-6-one [ka]

[0242] The synthesis method was the same as for 45a, except that 2-amino-4-bromobenzonitrile and propylenediamine were used as starting materials. LCMS (ESI): m / z 296.0 [M+H] + ;RT=0.715min (2.50min).

[0243] Intermediate 68a: N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride [ka]

[0244] The synthesis method is the same as that of Intermediate 2a, except that step 1 is omitted and methyl 5-bromopyridine-2-carboxylate and 4-Boc piperazine are coupled together. LCMS (ESI): m / z 221.2 [M+H] + ;RT=0.285min(6.00min).

[0245] Intermediate 69a: 2-chloro-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide hydrochloride [ka]

[0246] The synthesis method is similar to that of Intermediate 36a, except that N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester was used as the starting material. LCMS (ESI): m / z 252.2 [M+H] + ;RT=0.95min(3.00min).

[0247] Intermediate 70a: 6-chloro-N-methyl-5-(piperidin-4-yl)pyridinecarboxamide hydrochloride [ka]

[0248] 69a (240 mg, 0.85 mmol) and platinum dioxide (30 mg) were added sequentially to methanol (5 mL) at room temperature, and the mixture was stirred under hydrogen gas at room temperature for 4 hours. The reaction mixture was filtered, and the filtrate was concentrated to give 70a (130 mg, white solid). LCMS (ESI): m / z 254.3 [M+H] + ;RT=0.97min(3.00min).

[0249] Intermediate 71a: (R)-6-chloro-N-methyl-5-(3-methylpiperazin-1-yl)picolinamide [ka]

[0250] The synthesis method is similar to that of Intermediate 36a, except that (S)-1-N-Boc-2-methylpiperazine was used as the starting material. LCMS (ESI): m / z 269.2 [M+H] + ;RT=0.91min (3.0min).

[0251] Intermediate 72a: 6-chloro-N-(2,2-difluoroethyl)-5-(piperazin-1-yl)picolinamide [ka]

[0252] The synthesis method is the same as that of Intermediate 36a, except that 2,2-difluoroethan-1-amine was used as the starting material. LCMS (ESI): m / z 305.2 [M+H] + ;RT=1.02min (3.0min).

[0253] Intermediate 73a: 6-(difluoromethyl)-N-methyl-5-(piperazin-1-yl)picolinamide [ka]

[0254] The synthesis method is similar to that of Intermediate 36a, except that 5-bromo-6-(difluoromethyl)-N-methylpicolinamide was used as the starting material. LCMS (ESI): m / z 271.3 [M+H] + ;RT=1.54min (3.0min).

[0255] Intermediate 74a: N-methyl-5-(((2R,3S)-2-methylazetidin-3-yl)oxy)picolinamide [ka]

[0256] The synthesis method was the same as that of Intermediate 12a, except that (2R,3R)-3-hydroxy-2-methylazetidine-1-t-butylcarboxylate was used as the starting material. LCMS (ESI): m / z 222.2 [M+H] + ;RT=0.44min (3.0min).

[0257] Synthesis of compound 4 [ka]

[0258] 5a (150 mg, 0.48 mmol), 3a (124 mg, 0.48 mmol), and DIEA (187 mg, 1.45 mmol) were added to a 25 mL single-neck flask containing 5 mL of acetonitrile, and the mixture was reacted at 70°C for 2 hours. The reaction mixture was separated by pre-HPLC (formic acid) to obtain compound 4 (40 mg, white solid, yield: 19%).

[0259] LCMS(ESI): m / z 435.05[M+H] +;RT=3.198min(6.00min).

[0260] 1 H NMR(400MHz,DMSO-d6):δ11.30(s,1H),8.39(d,J=4.8Hz,1H),8.26(s,1H),8.19(s,1H),7.89(d,J=8.4Hz,1H),7.83(d,J=8.8Hz,1H),7.39(d, J=8.4Hz,1H),7.25(s,1H),7.08(d,J=3.2Hz,1H),6.73-6.71(m,1H),3. 66(s,2H),3.34-3.23(m,4H),2.78(d,J=4.8Hz,3H),2.64-2.54(m,4H).

[0261] Synthesis of Compound 5 - Compound 204 Compounds 5 to 204 were synthesized according to the method described for Compound 4, replacing Intermediates 5a and 3a with the intermediates shown in the table below. The structural formulas of the compounds in each example are shown in the table above.

[0262] [Table 4] TIFF2025538192000082.tif248170TIFF2025538192000083.tif243170TIFF2025538192000084.tif248170TIFF202 5538192000085.tif248170TIFF2025538192000086.tif248170TIFF2025538192000087.tif248170TIFF2025538192 000088.tif248170TIFF2025538192000089.tif248170TIFF2025538192000090.tif248170TIFF2025538192000091. tif248170TIFF2025538192000092.tif248170TIFF2025538192000093.tif243170TIFF2025538192000094.tif89170

[0263] [Table 5]

[0264] The present invention will be described in more detail below with reference to specific examples and data.It should be understood that these examples are merely illustrative examples for describing the present invention, and are intended to describe the specific combination of the present invention, preparation method, its function and effect, and are not intended to limit the scope of the present invention in any way.The beneficial effect of the drug combination of the present invention can be determined by other test models known to those skilled in the art.

[0265] Biological Examples Experimental Example 1: Evaluation of PARP1 / 2 inhibitory activity of compounds The PARP1 / 2 inhibitory activity of compounds of the present disclosure was tested in an assay using histone as a substrate.

[0266] Experimental objective: To determine the IC50 value of the compounds of the present application for inhibiting PARP1 / 2 enzyme activity according to established experimental methods. AZD-2281 (Olaparib) was used as a positive control compound.

[0267] Experimental Reagents:

[0268] Recombinant human PARP1 protein (Abcam, cat. ab279663); recombinant human PARP2 protein (BPS, cat. 80502); recombinant histone H1 (Active Motif, cat. 81126); NAD+, Biotin-Labeled (BPS, cat. 80610); SuperBlock (TBS) Blocking Buffer (Thermo Scientific™, cat. 37535); Streptavidin (HRP) (Abcam, cat. ab7403); LumiGLO® Peroxidase Chemiluminescent Substrate Kit (Seracare, cat. 5430-0040); 20xPBS (CST, cat. 9808S); 20xPBST (CST, cat. 9809S); AZD2281 (Selleck, cat. S1060).

[0269] Experimental method 1: PARP1 inhibitory activity 1. Compound preparation: Compounds were diluted in DMSO in a 384-well plate to a final concentration of 1000 times and prepared for use.

[0270] 2. Coating microplates: 1) Histones were diluted with PBS, and 25 μL of the histone mixture was added to each well and incubated for 2 hours. 2) Each well was washed five times with PBST solution. The solution was then wiped off with a clean paper towel. 3) 75 μL of blocking buffer was added to each well and incubated at room temperature for 1 hour. 4) Each well was washed five times with PBST solution. The solution was then wiped off with a clean paper towel.

[0271] 3. Ribosylation reaction: 1) 25 nL of compound (prepared for use) at a final concentration of 1000x was transferred to a 384-well reaction plate. 25 nL of 100% DMSO was added to each of the minimum and maximum control wells. 2) A PARP1 solution was prepared at a final concentration of 2.5x using 1x assay buffer. 3) 10 μL of enzyme solution was added to each of the compound wells and maximum control wells. 10 μL of 1x assay buffer was added to the minimum control well. 4) The plate was centrifuged at 1000 rpm for 60 seconds and incubated at room temperature for 15 minutes. 5) A substrate solution (prepared at a final concentration of 1.67x) was prepared in 1x assay buffer. 500 μM NAD+ was added to the substrate solution, and 15 μL of substrate solution was added to each well to initiate the reaction. 6) The plate was centrifuged at 1000 rpm for 60 seconds and incubated at room temperature for 2 hours. 7) Each well was washed five times with PBST solution. The solution was then wiped off with a clean paper towel.

[0272] 4. Detection: 1) Streptavidin-HRP solution was prepared and 25 μL was added to each well. The wells were centrifuged at 1000 rpm for 60 seconds and incubated at room temperature for 30 minutes. 2) Each well was washed five times with PBST solution and wiped with a clean paper towel. 3) 50 μL of ELISA chemiluminescent substrate was added to each well. 4) The wells were centrifuged at 1000 rpm for 60 seconds and read on EnSight after 5 minutes.

[0273] 5. Data Analysis The percent inhibition was calculated using the following formula: % inhibition = (maximum signal - compound signal) / (maximum signal - minimum signal) x 100, where "minimum signal" is the average value of the negative control wells and "maximum signal" is the average value of the positive control wells.

[0274] Dose-effect curve fitting: Dose-effect curves were fitted using log(inhibitor) vs. response-variable slope in the analytical software GraphPad Prism 5, with log concentration as the X-axis and percentage inhibition as the Y-axis, to obtain IC values ​​for inhibition of enzyme activity by compounds of the present disclosure. 50 The fitting equation was Y = Nadir + (Apex - Nadir) / (1 + 10^((logIC50-X)*HillSlope)).

[0275] Experimental method 2: PARP2 inhibitory activity The experimental method for PARP2 inhibitory activity was the same as Experimental Method 1, except that recombinant human PARP2 solution was used instead of the PARP1 solution used in Step 2 "2)" of "3. Ribosylation Reaction," and 500 μM NAD+ was not added in the fifth step of the ribosylation reaction. [Table 6] IC50 of inhibitory effect of compounds of the present disclosure on PARP1 / 2 enzymes [Table 6] TIFF2025538192000097.tif56170Note: " / " indicates no data found.

[0276] The results in Table 6 show that the compounds of the present invention have high selectivity for PARP1 and can reduce the toxicity caused by PARP2 without significantly reducing efficacy. The inventors also unexpectedly discovered that the compounds of the present disclosure have good physical and chemical stability, good bioavailability (e.g., low clearance rate), and good druggability. Therefore, the compounds of the present invention have fewer side effects than olaparib (AZD-2281) and are of high clinical value.

[0277] Example 2: MDA-MB-436 cell proliferation inhibition test Human breast cancer cells MDA-MB-436 (purchased from ATCC) were cultured in DMEM medium (supplemented with 10% fetal bovine serum and 1% double antibody) at 37°C and 5% carbon dioxide. Cells in the logarithmic growth phase were harvested and digested to prepare a cell suspension at a fixed concentration. The cell suspension was then inoculated into a 96-well plate. 100 μL of the cell suspension was added to each well of the 96-well plate and incubated overnight. Various concentrations of compounds were then added and the plate was placed in a cell culture incubator for 7 days. After incubation, 50 μL of CellTiter-Glo reagent was added to each well and mixed evenly for 2 minutes on a microplate shaker. After leaving the plate at room temperature for 60 minutes, the fluorescence value was measured using a Spark® multimode microplate reader and calculated using the formula: [(1-(RLU)] compound -RLU blank ) / (RLU control -RLU blank The cell growth inhibition rate was calculated according to the following formula: IC × 100%. GraphPad Prism 6.0 software was used. 50 The value was obtained.

[0278] Table 7. Inhibitory activity of compounds of the present disclosure against MDA-MB-436 cell proliferation [Table 7]

[0279] The experimental results showed that the compounds of the present invention have significant growth inhibitory activity against MDA-MB-436 cells.

[0280] Example 3: Evaluation of bidirectional permeability using the MDR1-MDCKII cell model 3.3 × 10 MDR1-MDCKII cells 5Cells were seeded into 96-well plates at a concentration of 100 cells / mL and grown for 4–7 days to form confluent monolayers. Test compounds were added at 2 μM to the wells on the apical or basolateral side of the monolayer and incubated for 2.5 hours at 37.0°C in a 5.0% CO2 incubator. The integrity of the monolayer was measured using a Lucifer Yellow exclusion assay. Buffer was removed from the apical and basolateral sides, and the test compound concentrations were measured by LC-MS / MS. The concentration data were used to calculate the apparent permeability of transport from the apical to the basolateral side of the monolayer, and the efflux ratio was calculated. The formula was: Efflux Ratio = Papp(BA) / Papp(AB). Table 8. Bidirectional permeability of compounds of the present disclosure to MDR1-MDCKII cells [Table 8]

[0281] The experimental results showed that the compounds of the present invention have high cell permeability and low efflux rate in MDR1-MDCKII cells.

[0282] Experimental Example 4: Preliminary Pharmacokinetic Study 1. Healthy ICR mice were administered the test compound by intravenous injection (1 mg / kg) and oral gavage (5 mg / kg). For each administration route, nine male mice weighing 30-35 g were randomly divided into three groups, with three mice in each group.

[0283] Animals were fasted for 12 h before the experiment and had free access to water. All mice were fed 4 h after administration.

[0284] 2. Blood collection time and sample processing Intravenous administration and oral gavage: 0.25 h, 0.5 h, 1.0 h, 2.0 h, 3.0 h, 4.0 h, 6.0 h, 8.0 h and 24 h after administration.

[0285] Blood was collected continuously from three animals at each time point. Plasma collection and processing: At the above-mentioned time points, 30-40 μL of venous blood was collected from the retroorbital plexus of each mouse, placed in EDTA-K2 tubes, and centrifuged at 3500 rpm for 10 min. The plasma was separated and frozen in a -20°C refrigerator.

[0286] 3. Sample Testing and Data Analysis The compound concentrations in mouse plasma were measured by LC / MS / MS. Pharmacokinetic parameters after administration were calculated using a non-compartmental model with Phoenix 8.3 software (Pharsight, USA).

[0287] 4. Experimental Results [Table 9] Pharmacokinetic parameters of the compounds of the present invention in mouse plasma [Table 9] NOTE: iv: intravenous injection (1 mg / kg), po: oral administration (5 mg / kg).

[0288] According to the experimental results, the compound of the present invention has a half-life (T 1 / 2 ) and showed a low clearance rate (CL), resulting in high bioavailability.

[0289] As can be seen from Table 6, the compounds of the present invention have high activity against PARP1 and high selectivity against PARP2, and can reduce the toxicity caused by PARP2 without significantly reducing the efficacy. As can be seen from Table 7, the compounds of the present invention have strong growth inhibitory activity against human breast cancer cells (MDA-MB-436). The inventors also unexpectedly discovered that the compounds of the present invention have higher cell permeability and lower efflux rates in MDR1-MDCKII cells, and also have good physical and chemical stability, good bioavailability (e.g., low clearance rate), and good druggability. Therefore, the compounds of the present invention have fewer side effects than olaparib (AZD-2281), and are of great value for clinical application.

Claims

1. A compound represented by general formula (I), or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof. 【Chemistry 1】 (where, X １ are independently -N-, -NR １４ -, -CR ７ -, -CR ７ R ７’ , -CH ２ CR ７ R ７’ -, -CR ７ R ７’ -CH ２ -, O, and S; X ２ are independently -N-, -NR １５ -, -CR ８ , -CR ８ R ８’ -, O, and S; X ３ are independently -N-, -NR １６ -, -CR ９ -, O, and S; X ４ , X ８ are each independently selected from —N— and —C—; 【Chemistry 2】 is a single bond or a double bond, and X １ , X ２ , X ３ , X ４ , X ８ together form a 5-membered heteroaryl, or a partially saturated 5- or 6-membered heterocyclyl, wherein said heteroaryl or heterocyclyl each independently contains 1, 2 or 3 heteroatoms independently selected from N, O or S; R ７ , R ７’ , R ８ , R ８’ , R ９ are each independently hydrogen, halogen, hydroxy, cyano, C １ ~C ３ Alkoxy, unsubstituted or substituted C ３ ~C ６ Cycloalkyl, or unsubstituted or substituted C １ ~C ６ alkyl, or R ７ and R ７’ or R ８ and R ８’ is jointly C ３ ~C ６ constitutes a cycloalkyl, and R ７ , R ７’ , R ８ , R ８’ , R ９ are each independently preferably hydrogen, halogen or C １ ~C ４ alkyl, and R ７ , R ７’ , R ８ , R ８’ , R ９ are each independently more preferably hydrogen, F, or methyl, or preferably R ７ and R ７’ or R ８ and R ８’ is jointly C ３ ~C ４ cycloalkyl, such as cyclopropyl or cyclopentyl; R １４ , R １５ , R １６ are each independently hydrogen, unsubstituted or substituted C ３ ~C ６ Cycloalkyl, unsubstituted or substituted C １ ~C ６ alkyl, and R １４ , R １５ , R １６ are each independently preferably hydrogen or C １ ~C ３ alkyl, and R １４ , R １５ , R １６ are each independently more preferably methyl; X ５ , X ６ are each independently —N— and —CR １０ -, and R １０ is hydrogen, halogen, cyano, unsubstituted or substituted C １ ~C ６ Alkoxy, unsubstituted or substituted C １ ~C ６ alkyl, and R １０ is preferably hydrogen, halogen, cyano, or C １ ~C ４ alkyl, and R １０ is more preferably hydrogen, fluorine, chlorine or methyl, X ７ is -N- or -CR １７ - and R １７ is hydrogen, halogen, cyano, unsubstituted or substituted C １ ~C ６ Alkoxy, unsubstituted or substituted C １ ~C ６ alkyl, and R １７ is preferably hydrogen, halogen, cyano, or C １ ~C ４ alkyl, and R １７ is more preferably hydrogen, fluorine, chlorine or methyl, R １ , R １’ , R ２ , R ３ , R ４ , R ５ are each independently hydrogen, unsubstituted or substituted C １ ~C ６ alkyl, or R ４ , R ５ are C together with the carbon atoms to which they are attached. ３ ~C ６ constitutes a cycloalkyl, s and n are each independently selected from 0, 1, and 2; Y is N or CH; R ６ teeth, 【Transformation 3】 is selected from Each R １１ are independently halogen, cyano, C １ ~C ３ Alkoxy, carbonyl, -CONHR １３ , amino, preferably halogen, —CONHR １３ and cyano, more preferably -CONHR １３ is selected from m is 0, 1, 2, or 3; R １２ is hydrogen, cyano, halogen, unsubstituted or substituted C １ ~C ４ alkyl, R １３ is hydrogen, unsubstituted or substituted C １ ~C ６ Alkyl, unsubstituted or substituted C ３ ~C ８ Cycloalkyl, unsubstituted or substituted C １ ~C ６ alkoxy, or unsubstituted or substituted 3- to 8-membered heterocycloalkyl, preferably R １３ is hydrogen, unsubstituted or halogen-substituted C １ ~C ４ Alkyl, C ３ ~C ６ cycloalkyl, or C １ ~C ６ The heterocycloalkyl is an alkoxy group, and the heterocycloalkyl group contains 1 to 3 heteroatoms selected from N, O, and S. Preferably, R １３ is methyl, ethyl, C ２ ~C ３ alkoxy, cyclopropanyl, propylene oxide, oxetanyl, or oxiranyl; Here, R １ , R １’ , R ２ , R ３ , R ４ , R ５ , R ７ , R ７’ , R ８ , R ８’ , R ９ , R １０ , R １２ , R １３ , R １４ , R １５ , R １６ , R １７ The substitution in １ ~C ４ Alkyl, C １ ~C ６ Alkoxy, halogen, hydroxy, cyano, amino, carboxy, C ３ ~C ６ cycloalkyl; however, X ５ and X ８ When both of X are —N—, １ , X ２ , X ３ , X ４ at least one of is —N—; X ４ and X ５ When both of X are —N—, ３ is also -N-, X ３ is oxygen and X １ , X ２ , X ４ , X ８ is -C- and X ７ Ga-CR １７ -, then R １ , R １’ , R １７ is not hydrogen at the same time, Preferably, however, X ８ When is -N-, X １ , X ２ , X ３ At least one of the groups is —N—.

2. In general formula (I), 【Chemistry 4】 is selected from the following structures: or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate, or prodrug thereof. 【Transformation 5】 (Here, the definitions of each substituent are the same as in claim 1.)

3. In general formula (I), 【Transformation 6】 is selected from the following structures: or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate, or prodrug thereof. 【Transformation 7】 【change】 (Here, the definitions of each substituent are the same as in claim 1.)

4. In general formula (I), 【Transformation 8】 is selected from the following structures: or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate, or prodrug thereof. 【Chemistry 9】 (Here, the definitions of each substituent are the same as in claim 1.)

5. In general formula (I), 【Chemistry 10】 is selected from the following structures: or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate, or prodrug thereof. 【Chemistry 11】 (Here, the definitions of each substituent are the same as in claim 1.)

6. 6. The compound of any one of claims 1 to 5, wherein the compound of formula (I) is selected from the following specific compounds: Table 1

7. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) according to any one of claims 1 to 6, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, and a pharmaceutically acceptable carrier.

8. Use of a compound of formula (I) according to any one of claims 1 to 6, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate or prodrug thereof, or a pharmaceutical composition according to claim 7, in the preparation of a medicament for preventing or treating a disease ameliorated by inhibition of PARP1.

9. the disease is cancer, and the genome of the cancer is homologous recombination repair deficient; Alternatively, the cancer is dependent on a DNA double-strand break and defective homologous recombination repair pathway; Alternatively, the cancer comprises one or more cancer cells, wherein the cancer cells are deficient in the ability to repair DNA double-strand breaks by homologous recombination compared to normal cells; Alternatively, the cancer comprises one or more cancer cells, wherein the cancer cells are BRCA1 or BRCA2 deficient or have a type of BRCA1 or BRCA2 mutation.

10. The use of claim 9, wherein the cancer includes, but is not limited to, malignant tumors such as ovarian cancer, breast cancer, fallopian tube cancer, endometrial cancer, peritoneal cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, biliary tract cancer, osteosarcoma, cervical cancer, head and neck tumors, germ cell cancer and embryonal cancer, esophageal cancer, malignant glioma, Ewing's sarcoma, pancreatic cancer, melanoma, bile duct cancer, prostate cancer, small cell lung cancer, non-small cell lung cancer, lymphoma, and blood cancer.

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