Compound containing indoline structure as well as preparation method and application thereof

Abstract

The invention relates to a compound containing an indoline structure as well as a preparation method and application thereof, and belongs to the technical field of medicines. The invention particularly relates to a compound containing an indoline structure as shown in a general formula I, a stereoisomer and pharmaceutically acceptable salt of the compound, a preparation method of the compound and the stereoisomer and the pharmaceutically acceptable salt, and application of the compound and the stereoisomer and the pharmaceutically acceptable salt to preparation of drugs for treating PD-1 / PD-L1 protein / protein interaction and NAMPT related diseases. The compound has a novel chemical structure, has high-level inhibitory activity on PD-1 / PD-L1 protein / protein interaction and NAMPT, can significantly inhibit tumor cell growth, and can be used for preparing drugs for treating diseases related to PD-1 / PD-L1 protein / protein interaction and NAMPT, such as cancer or virus infection. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a compound containing an indoline structure, a preparation method thereof and an application thereof, belonging to the technical field of medicine. Background Art

[0002] In recent years, the immune checkpoint blockade therapy based on programmed cell death-1 (PD-1) and its ligand PD-L1 has achieved great success in the clinical treatment of tumors. As a T cell surface receptor, when PD-1 binds to PD-L1, it generates a negative immune regulation signal, resulting in the inhibition of T cell activation and proliferation, and the reduction of cytokine secretion such as interleukin-2 (IL-2) and interferon-γ (IFN-γ). In the tumor microenvironment, the expression of PD-1 on infiltrating T cells is up-regulated, and the expression of PD-L1 on tumor cells is highly expressed, thus activating the PD-1 / PD-L1 signaling pathway, causing the function of tumor-specific CD8 + T cells to be damaged, leading to tumor immune escape. Therefore, blocking the PD-1 / PD-L1 signaling pathway can reverse the above immune suppression mechanism, reshape the body's immune system's recognition and killing ability of tumor cells, and open up a new effective way for tumor treatment. At present, more than a dozen anti-PD-1 / PD-L1 monoclonal antibody drugs such as pembrolizumab of Merck Sharp & Dohme and nivolumab of Bristol-Myers Squibb have been approved for marketing, and have achieved good clinical efficacy in the treatment of various tumors such as melanoma, non-small cell lung cancer, gastric cancer, and urothelial cancer. With the advancement of clinical research, more indications have been approved.

[0003] However, monoclonal antibody drugs also have certain limitations. Their preparation and purification processes are complex, costly, and easily degraded by proteases, making it difficult to administer them through the oral route. At the same time, they may cause serious side effects due to their immunogenicity. In contrast, small molecule compounds show greater flexibility in regulating pharmacokinetic properties and have great potential in optimizing production and dosage forms. In addition, primary and secondary drug resistance that occur in PD-1 / PD-L1 monoclonal antibody therapy are bottleneck problems in this field, which greatly limit the application of such immunotherapy drugs.

[0004] Nicotinamide adenine dinucleotide (NAD + ) is one of the important coenzymes in cellular redox reactions and plays a key role in processes such as energy metabolism, gene expression regulation, DNA repair, cell signal transduction, and apoptosis. The level and metabolism of NAD + are strictly regulated, and its synthesis mainly occurs through the following several pathways: de novo synthesis from tryptophan; synthesis beside nicotinic acid and nicotinamide ribosome; recycling of NAD+ The metabolite NAM is converted to NMN, which then undergoes salvage synthesis. Salvage synthesis is an important pathway for NAD + biosynthesis, and nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme in this pathway.

[0005] Studies have shown that tumor cells consume NAD + at a faster rate than normal cells, with abnormally active metabolism. NAMPT and NAD + are upregulated in a variety of human malignancies and play a key role in the occurrence, development, and recurrence of cancer. In addition, unlike tumor cells that mainly rely on NAMPT, normal cells can synthesize NAD + through other biosynthetic pathways. Therefore, inhibiting the function of NAMPT has become a specific anti-tumor treatment strategy, and representative NAMPT inhibitors include CHS-828 and FK866. However, due to insufficient clinical efficacy and the problem of dose-limiting toxicity, the related clinical studies of the two drugs have been forced to terminate.

[0006]

[0007] Studies have shown that overexpressed NAMPT in tumor cells promotes the proliferation of immunosuppressive cells MDSCs, thereby promoting the formation of an immunosuppressive microenvironment. At the same time, the continuous accumulation of lactic acid produced by tumor cell glycolytic metabolism can promote the differentiation of monocytes into dendritic cells. The differentiated cells inhibit the T cell immune response by secreting immunosuppressive factors, thus directly weakening the immune response. It can be seen that the high metabolic state of tumor cells triggers the formation of an immunosuppressive microenvironment, which is one of the important reasons for tumor cells to develop resistance to anti-PD-1 / PD-L1 therapy. Research has shown that inhibiting NAMPT can upregulate the expression of PD-L1 through SIRT-mediated NF-κB p65 acetylation regulation, while inhibiting PD-L1 can activate the glycolytic pathway in a HIF-1α-dependent manner to induce the expression of NAMPT (Yang Y et al. The regulatory relationship between NAMPT and PD-L1 in cancer and identification of a dual-targeting inhibitor. EMBO molecular medicine 2024, 16(4): 885-903). Therefore, simultaneously inhibiting the PD-1 / PD-L1 protein / protein interaction and NAMPT has the potential to synergistically activate the immune function and is expected to solve the problems of PD-1 / PD-L1 inhibitor resistance and NAMPT inhibitor dose-limiting toxicity to a certain extent. Therefore, the targeted development of dual inhibitors of PD-1 / PD-L1 interaction and NAMPT is expected to obtain compounds with outstanding anti-tumor activity and unique mechanisms of action, which has important research significance. Summary of the Invention

[0008] The present invention provides a compound containing an indoline structure, a preparation method thereof and an application. Research has shown that such compounds can significantly inhibit the PD-1 / PD-L1 protein / protein interaction and NAMPT activity. The present invention relates to a compound containing an indoline structure represented by general formula I, its stereoisomers and pharmaceutically acceptable salts, a preparation method thereof, and a pharmaceutical composition containing the compound, wherein the meanings of substituents R1, R2, R3, as well as X, m and n are all described in the specification. The present invention relates to the fact that the compound has a high level of inhibitory activity against both the PD-1 / PD-L1 protein / protein interaction and NAMPT. The compound containing an indoline structure, its stereoisomers and pharmaceutically acceptable salts can be used to prepare drugs for preventing and / or treating diseases related to the PD-1 / PD-L1 protein / protein interaction and NAMPT, such as cancer or viral infection.

[0009] The present invention relates to a compound containing an indoline structure represented by general formula I, its stereoisomers, and pharmaceutically acceptable salts,

[0010]

[0011] wherein,

[0012] X is selected from CH or N;

[0013] R1 is selected from hydrogen, methyl or halogen;

[0014] R2 is selected from

[0015] R4 and R5 are each independently selected from hydrogen, (C1–C4)alkyl or hydroxy(C1–C4)alkyl, or R4, R5 and the nitrogen atom to which they are attached together form a 4- to 6-membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may be optionally substituted with 1 to 3 R7;

[0016] R6 is independently selected from hydrogen, (C1–C4)alkyl, (C1–C4)alkylcarbonyl or (C1–C4)alkylsulfonyl;

[0017] R7 is independently selected from hydrogen, halogen, hydroxy, carboxy, (C1–C4)alkyl or hydroxy(C1–C4)alkyl;

[0018] m is an integer selected from 1 to 3;

[0019] n is an integer selected from 0 to 5, wherein,

[0020] when n is 0, R3 is selected from: when n is 1 to 5, R3 is selected from:

[0021] The present invention preferably relates to a compound containing an indoline structure represented by general formula I, its stereoisomers, and pharmaceutically acceptable salts, wherein,

[0022] X is selected from CH or N;

[0023] R1 is selected from hydrogen, methyl, fluorine or chlorine;

[0024] R2 is selected from

[0025] R4 and R5 are each independently selected from hydrogen or (C1–C4)alkyl, or R4, R5 and the nitrogen atom to which they are attached together form a 4- to 6-membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may be optionally substituted with 1 to 3 R7;

[0026] R6 is independently selected from hydrogen or (C1–C4)alkyl;

[0027] R7 is independently selected from hydrogen, halogen, hydroxyl, carboxyl or hydroxy(C1–C4)alkyl;

[0028] m is an integer selected from 1 to 3;

[0029] n is an integer selected from 0 to 5, where,

[0030] when n is 0, R3 is selected from: when n is 1 to 5, R3 is selected from:

[0031] The present invention more preferably relates to a compound containing an indoline structure of general formula I, its stereoisomers and pharmaceutically acceptable salts, where,

[0032] X is selected from CH or N;

[0033] R1 is selected from hydrogen, methyl, fluorine or chlorine;

[0034] R2 is selected from

[0035] selected from:

[0036] selected from:

[0037] m is an integer selected from 1 to 3;

[0038] n is an integer selected from 0 to 5, where,

[0039] when n is 0, R3 is selected from: when n is 1 to 5, R3 is selected from:

[0040] A compound containing an indoline structure of general formula I of the present invention, its stereoisomers and pharmaceutically acceptable salts are preferably selected from the following compounds, but these compounds do not imply any limitation to the present invention:

[0041]

[0042]

[0043]

[0044]

[0045] In addition, the present invention also includes prodrugs of the compounds of the present invention. The prodrugs of the compounds of the present invention are derivatives of general formula I, which may themselves have weak activity or even no activity, but after administration, they are converted into the corresponding bioactive forms under physiological conditions (such as by metabolism, solvolysis or other means).

[0046] The indoline structure-containing compounds of general formula I as described above, their stereoisomers and pharmaceutically acceptable salts, wherein the pharmaceutically acceptable salts include salts formed with inorganic acids, organic acids or alkali metal ions; the inorganic acids are selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid or phosphoric acid; the organic acids are selected from: succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid or p-toluenesulfonic acid; the alkali metal ions are selected from lithium ions, sodium ions or potassium ions.

[0047] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to straight-chain or branched-chain alkyl. Represents the substituent connection point.

[0048] The present invention can use the indoline structure-containing compounds of general formula I as described above, their stereoisomers and pharmaceutically acceptable salts as active ingredients, and mix them with pharmaceutically acceptable carriers or excipients to prepare a composition. The carriers or excipients include diluents, binders, wetting agents, disintegrants, lubricants, glidants, etc. well known in the art. Diluents include but are not limited to starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, calcium hydrogen phosphate, etc.; wetting agents include but are not limited to water, ethanol, isopropanol, etc.; binders include but are not limited to starch paste, dextrin, syrup, honey, glucose solution, acacia mucilage, gelatin mucilage, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, polyethylene glycol, etc.; disintegrants include but are not limited to dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium dodecyl sulfate, etc.; lubricants and glidants include but are not limited to talc, silica, polyethylene glycol, etc.

[0049] The pharmaceutical composition of the present invention can be formulated into several dosage forms, including but not limited to injections, tablets, capsules, etc.

[0050] The indoline structure-containing compounds, their stereoisomers and pharmaceutically acceptable salts provided by the present invention can be used in combination with other active ingredients to achieve better therapeutic effects.

[0051] The present invention also provides the use of a compound containing an indoline structure of general formula I, its stereoisomers, and pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing and / or treating diseases related to PD-1 / PD-L1 protein / protein interaction and NAMPT.

[0052] Furthermore, the diseases related to PD-1 / PD-L1 protein / protein interaction and NAMPT are selected from cancer or infectious diseases.

[0053] Still further, the cancer is selected from lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell cancer, glioma, neuroblastoma, squamous cell lung cancer, adenocarcinoma of the lung, bladder cancer, gastric cancer, colon cancer, colorectal cancer, kidney cancer, cholangiocarcinoma, gastric cancer, esophageal squamous cell cancer, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, skin cancer, epithelial cell carcinoma, leukemia or cervical cancer; the infectious disease is selected from bacterial infection or viral infection.

[0054] Advantages of the present invention: The compound containing an indoline structure provided by the present invention has a novel chemical structure. The compound has significant inhibitory activity against PD-1 / PD-L1 protein / protein interaction and NAMPT in in vitro biological activity studies, and it can significantly inhibit the growth of tumor cells and can be used for the treatment and prevention of various diseases such as cancer. Detailed implementation mode

[0055] The following synthetic routes summarize and describe the preparation of derivatives of general formula I of the present invention. All raw materials are prepared by the methods described in these processes or are commercially available by methods well-known to those of ordinary skill in the field of organic chemistry. The derivatives of the present invention are all prepared by the methods described in these processes or by methods similar thereto, which are well-known to those of ordinary skill in the field of organic chemistry. All variable factors applied in these processes are defined as in the claims.

[0056] When n = 1 - 5 in general formula I, the preparation method of the compound containing an indoline structure includes the following steps:

[0057]

[0058] (a) Using 4-arylindole as a raw material, intermediate 2 is prepared under the action of a reducing agent such as sodium cyanoborohydride.

[0059] (b) Using intermediate 2 as a raw material, intermediate 3 is prepared through an aromatic nucleophilic substitution reaction with 7-bromo-4-chloropyrido[3,2-d]pyrimidine or 3-bromo-8-chloro-1,7-naphthyridine under acid catalysis.

[0060] (c) Using intermediate 3 as a raw material, reacting with pinacol vinyl borate or tributylvinyltin reagent through a coupling reaction to obtain intermediate 4;

[0061] (d) Using intermediate 4 as a raw material, reacting with a small molecule halide through a nucleophilic substitution reaction or reacting with an alcohol compound through a Mitsunobu reaction to obtain intermediate 5;

[0062] (e) Using intermediate 5 as a raw material, reacting under the action of an osmium reagent and an oxidant to obtain intermediate 6;

[0063] (f) Using intermediate 6 as a raw material, reacting with an amine compound under the action of sodium cyanoborohydride or sodium triacetoxyborohydride through a reductive amination reaction to obtain the target compound in general formula I (n = 1 - 5);

[0064] Alternatively, when n = 1 - 5 in general formula I, the preparation method of the indoline structure compound comprises the following steps:

[0065]

[0066] (g) Using intermediate 6 as a raw material, reacting with a mono - Boc - alkyl diamine compound under the action of sodium cyanoborohydride or sodium triacetoxyborohydride to obtain intermediate 7;

[0067] (h) Using intermediate 7 as a raw material, removing the protecting agent under acidic conditions to obtain intermediate 8;

[0068] (i) Using intermediate 8 as a raw material, obtaining the target compound in general formula I (n = 1 - 5) through a nucleophilic substitution or amidation reaction;

[0069] When n = 0 in general formula I, the preparation method of the indoline structure compound comprises the following steps:

[0070]

[0071] (j) Using intermediate 6 as a raw material, reacting under the action of a reducing agent such as sodium borohydride to obtain intermediate 9;

[0072] (k) Using intermediate 9 as a raw material, reacting under the action of a chlorinating agent such as thionyl chloride to obtain intermediate 10;

[0073] (l,m) Using intermediate 10 as a raw material, obtaining intermediate 12 through a Gabriel reaction;

[0074] (n) Using intermediate 12 as a raw material, obtaining the target compound in general formula I (n = 0) through a nucleophilic substitution or amidation reaction;

[0075] Alternatively, when n = 0 in general formula I, the preparation method of the indoline structure compound comprises the following steps:

[0076]

[0077] (o) Using intermediate 3 as a raw material, reacting with a cyanide reagent such as cuprous cyanide or zinc cyanide through a coupling reaction to obtain intermediate 13;

[0078] (p) Using intermediate 13 as a raw material, reacting with a small molecule halide through a nucleophilic substitution reaction or reacting with an alcohol compound through a Mitsunobu reaction to obtain intermediate 14;

[0079] (q) Using intermediate 14 as a raw material, reducing it with borane or through a catalytic hydrogenation reaction to obtain intermediate 12;

[0080] (r) Using intermediate 12 as a raw material, obtaining the target compound in general formula I (n = 0) through a nucleophilic substitution or amidation reaction.

[0081] The definitions of R1, R2, R3, X, m, and n are as described in the claims. The compounds with an indoline structure having general formula I according to the present invention can all be prepared by the methods described in the above reaction routes or similar methods.

[0082] The following non-limiting examples can enable those of ordinary skill in the art to more comprehensively understand the present invention, but do not limit the present invention in any way.

[0083] Example 1

[0084] N-(2-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-1)

[0085]

[0086] Step 1: 3-(1H-Indol-4-yl)phenol

[0087]

[0088] 4-Bromoindole (15 g, 76.5 mmol), 3-Hydroxybenzeneboronic acid (12.7 g, 91.8 mmol), Pd(dppf)Cl2 (1.12 g, 1.53 mmol), and potassium carbonate (26.4 g, 191.3 mmol) were successively added to a mixed solution of 1,4-dioxane (120 mL) and water (30 mL). The reaction was carried out at 90 °C for 3 h under N2 protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate several times. The filtrate was poured into 200 mL of water and extracted with ethyl acetate (300 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product as a pale yellow solid, which was used directly in the next step without purification. 1 H NMR (400 MHz, Chloroform-d) δ 8.16 (s, 1H), 7.34–7.24 (m, 2H), 7.23–7.18 (m, 2H), 7.17–7.15 (m, 1H), 7.13–7.06 (m, 2H), 6.79–6.74 (m, 1H), 6.68–6.64 (m, 1H). ESI-MS m / z: 210.1 [M+H] + 。

[0089] Step 2: 3-(Indolin-4-yl)phenol

[0090]

[0091] At room temperature, 3-(1H-Indol-4-yl)phenol (14.8 g, 76.5 mmol) was added to 40 mL of glacial acetic acid, and then NaBH3CN (9.62 g, 153.0 mmol) was slowly added under an ice bath. After addition, the reaction was carried out at room temperature for 2 h. After the reaction was completed, the reaction solution was poured into 400 mL of water, and the pH was adjusted to neutral with 40% NaOH solution under an ice bath. It was extracted with ethyl acetate (300 mL × 3), the organic layers were combined and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was separated and purified by column chromatography to obtain 13.8 g of a white solid, and the total yield of the two steps was 85.4%. 1 H NMR (400 MHz, Chloroform-d) δ 7.22–7.17 (m, 1H), 7.05–7.00 (m, 1H), 6.94–6.90 (m, 1H), 6.75–6.67 (m, 3H), 6.64–6.60 (m, 1H), 3.47 (t, J = 8.2 Hz, 2H), 3.01 (t, J = 8.2 Hz, 2H). ESI-MS m / z: 212.2 [M+H] + 。

[0092] Step 3: 3-(1-(7-Bromopyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol

[0093]

[0094] Add 3-(1H-Indol-4-yl)phenol (8 g, 37.9 mmol) and 7-bromo-4-chloropyrido[3,2-d]pyrimidine (9.26 g, 37.9 mmol) into a 500 mL eggplant-shaped flask, add 200 mL of isopropanol, then add 1 drop of concentrated hydrochloric acid, and react at 70 °C for 1 h. A large amount of yellow solid precipitates. After the reaction is completed, cool the reaction solution to room temperature, filter it by suction, wash the filter cake with ether, and dry it to obtain 15.5 g of yellow solid, with a yield of 97.6%. 1 1H NMR (600 MHz, DMSO-d6) δ 9.10 (d, J = 2.2 Hz, 1H), 8.95 (s, 1H), 8.62 (d, J = 2.2 Hz, 1H), 8.53 (d, J = 8.2 Hz, 1H), 7.46–7.40 (m, 1H), 7.33–7.21 (m, 3H), 6.95–6.90 (m, 2H), 6.84–6.79 (m, 1H), 4.95 (t, J = 7.7 Hz, 2H), 3.31 (t, J = 7.7 Hz, 2H). ESI-MS m / z: 419.0 [M+H] + 。

[0095] Step 4: 3-(1-(7-Vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol

[0096]

[0097] Dissolve 3-(1-(7-bromopyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol (15 g, 35.8 mmol) in a mixed solution of 140 mL of DMF and 17 mL of water. Subsequently, add vinylboronic acid pinacol ester (6.61 g, 42.9 mmol), potassium carbonate (12.4 g, 89.4 mmol), and Pd(PPh3)4 (0.83 g, 0.72 mmol) in sequence, and react at 100 °C for 2 h under N2 protection. After the reaction is completed, cool the reaction solution to room temperature, then pour the reaction solution into 1500 mL of water. A large amount of yellow solid precipitates. Filter it by suction and dry the filter cake to obtain 11.6 g of yellow solid, with a yield of 88.5%. 11H NMR (600 MHz, Chloroform-d) δ 8.83 (d, J = 2.1 Hz, 1H), 8.66 (s, 1H), 8.38 (d, J = 8.2 Hz, 1H), 8.17 (s, 1H), 7.49–7.46 (m, 1H), 7.28–7.25 (m, 1H), 7.25–7.21 (m, 1H), 7.07 (d, J = 7.5 Hz, 1H), 6.91–6.88 (m, 2H), 6.84–6.81 (m, 1H), 6.80–6.76 (m, 1H), 6.04 (d, J = 17.7 Hz, 1H), 5.57 (d, J = 11.0 Hz, 1H), 4.86 (t, J = 7.9 Hz, 2H), 3.15 (t, J = 7.9 Hz, 2H). ESI-MS m / z: 367.2 [M+H] + 。

[0098] Step 5: 4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine

[0099]

[0100] 3-(1-(7-Vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol (8 g, 21.8 mmol), 1-(3-chloropropyl)pyrrolidine (3.87 g, 26.2 mmol), and potassium carbonate (9.05 g, 65.5 mmol) were successively added to 80 mL of DMF, and the mixture was stirred at 70 °C for 18 h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with 100 mL of ethyl acetate, and then filtered through diatomaceous earth. The filter cake was washed with ethyl acetate several times. The filtrate was poured into 300 mL of water, and the mixture was extracted with ethyl acetate (200 mL × 3). The organic phase was washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The crude product was separated and purified by column chromatography to obtain 8.6 g of a yellow solid with a yield of 82.5%. 11H NMR (600 MHz, Chloroform-d) δ 8.80 (d, J = 2.2 Hz, 1H), 8.70 (s, 1H), 8.46 (d, J = 8.2 Hz, 1H), 7.99 (d, J = 2.2 Hz, 1H), 7.28 (t, J = 7.9 Hz, 1H), 7.21–7.17 (m, 2H), 6.93–6.88 (m, 2H), 6.84–6.78 (m, 2H), 6.01 (d, J = 17.7 Hz, 1H), 5.52 (d, J = 11.0 Hz, 1H), 4.94–4.81 (m, 2H), 4.15–4.06 (m, 2H), 3.05 (s, 1H), 2.91 (s, 1H), 2.80–2.55 (m, 6H), 2.16–2.05 (m, 2H), 1.84–1.75 (m, 4H). ESI-MS m / z: 478.4 [M+H] + 。

[0101] Step 6: 4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0102]

[0103] Dissolve the intermediate 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine (8 g, 16.8 mmol) in a mixed solution of 1,4-dioxane (200 mL) and water (50 mL). Subsequently, add K2OsO4·2H2O (0.25 g, 0.67 mmol), stir at room temperature for 30 min, then place the reaction solution in an ice bath, add NaIO4 (9.77 g, 45.7 mmol), and react at no more than 10 °C for 6 h. After the reaction is completed, perform suction filtration. Wash the filter cake with dichloromethane until it is white. Pour the filtrate into 400 mL of 4 M sodium thiosulfate solution, stir for 30 min, extract with dichloromethane (400 mL × 3). Dry the organic phase over anhydrous sodium sulfate, perform suction filtration, evaporate the filtrate to dryness, and purify the residue by column chromatography to obtain 5.9 g of a yellow solid with a yield of 73.5%. 11H NMR (600 MHz, Chloroform-d) δ 10.31 (s, 1H), 9.24 (d, J = 2.1 Hz, 1H), 8.86 (s, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.51 (d, J = 8.2 Hz, 1H), 7.39–7.34 (m, 2H), 7.16 (dd, J = 7.6, 1.0 Hz, 1H), 7.04–7.01 (m, 1H), 6.99 (dd, J = 2.5, 1.6 Hz, 1H), 6.92 (ddd, J = 8.3, 2.6, 0.9 Hz, 1H), 4.97 (t, J = 8.1 Hz, 2H), 4.09 (t, J = 6.4 Hz, 2H), 3.31 (t, J = 8.0 Hz, 2H), 2.71 (t, J = 7.5 Hz, 2H), 2.61 (s, 4H), 2.10–2.05 (m, 2H), 1.84–1.81 (m, 4H). ESI-MS m / z: 480.4 [M+H] + 。

[0104] Step 7: tert-Butyl (2-(imidazo[1,2-a]pyridine-6-carboxamido)ethyl)carbamate

[0105]

[0106] Imidazo[1,2-a]pyridine-6-carboxylic acid (2 g, 12.3 mmol), N-Boc-ethylenediamine (1.8 g, 11.2 mmol), and HATU (5.54 g, 14.6 mmol) were successively added to 30 mL of dichloromethane, and the mixture was stirred at room temperature for 10 min. DIPEA (4.35 g, 33.6 mmol) was slowly added, and after addition, the reaction was continued for 1 h. After completion of the reaction, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to obtain 2.74 g of a white solid with a yield of 80.3%. 1 1H NMR (600 MHz, Chloroform-d) δ 8.19–8.14 (m, 2H), 7.96 (s, 1H), 7.77–7.74 (m, 1H), 7.66 (s, 1H), 7.35–7.32 (m, 1H), 5.26 (s, 1H), 3.60 (q, J = 5.2 Hz, 2H), 3.44 (q, J = 5.8 Hz, 2H), 1.43 (s, 9H). ESI-MS m / z: 305.2 [M+H] + 。

[0107] Step 8: Imidazo[1,2-a]pyridine-6-carboxamide, N-(2-aminoethyl)-, hydrochloride

[0108]

[0109] Dissolve tert-butyl (2-(imidazo[1,2-a]pyridine-6-carboxamido)ethyl)carbamate (2 g, 6.57 mmol) in 20 mL of methanol, then add 4 mL of hydrochloric acid dioxane solution, and react at room temperature for 3 h. After the reaction is completed, evaporate the solvent under reduced pressure to obtain 1.58 g of white solid, with a yield of 100%. ESI-MS m / z: 205.1 [M+H] + 。

[0110] Step 9: N-(2-(((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-1)

[0111]

[0112] Dissolve N-(2-aminoethyl)imidazo[1,2-a]pyridine-6-carboxamide hydrochloride (90.3 mg, 0.38 mmol) in 4 mL of methanol, add NaOH (30 mg, 0.75 mmol), and react at room temperature for 20 min. Slowly adjust the pH of the reaction system to neutral with a methanol solution of glacial acetic acid, then add 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl))pyrido[3,2-d]pyrimidine-7-carbaldehyde (120 mg, 0.25 mmol), then add 2 mL of dichloromethane, stir at room temperature for 30 min, add NaBH3CN (39.3 mg, 0.63 mmol), and continue to react at room temperature for 2 h. After the reaction is completed, pour the reaction solution into 20 mL of water, extract with dichloromethane (15 mL×3), combine the organic phases, concentrate under reduced pressure, and purify the crude product by column chromatography to obtain 70 mg of yellow solid, with a yield of 41.9%. 11H NMR (600 MHz, Chloroform-d) δ 8.76 (s, 1H), 8.70 (d, J = 2.2 Hz, 1H), 8.68 (s, 1H), 8.36 (d, J = 8.2 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.61 (s, 2H), 7.55 (d, J = 9.4 Hz, 1H), 7.39 (d, J = 1.8 Hz, 1H), 7.27 (t, J = 7.9 Hz, 2H), 7.04 (d, J = 7.6 Hz, 1H), 6.97–6.89 (m, 3H), 6.83 (dd, J = 8.3, 2.6 Hz, 1H), 4.82 (t, J = 8.1 Hz, 2H), 4.03–4.00 (m, 2H), 3.98 (s, 2H), 3.55 (q, J = 5.6 Hz, 2H), 3.18 (t, J = 8.1 Hz, 2H), 2.91 (t, J = 5.7 Hz, 2H), 2.70 (t, J = 7.6 Hz, 2H), 2.61 (s, 4H), 2.07–1.99 (m, 2H), 1.80–1.77 (m, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.75, 159.17, 157.96, 154.77, 149.11, 147.81, 145.04, 144.82, 141.73, 141.48, 138.25, 134.79, 133.80, 132.27, 131.07, 130.05, 128.80, 127.54, 124.10, 123.35, 120.91, 120.32, 117.83, 116.42, 114.71, 114.68, 113.88, 66.27, 54.59, 54.06, 52.66, 50.15, 48.37, 29.07, 28.52, 23.53. ESI-MS m / z: 668.1 [M+H] + 。

[0113] Example 2

[0114] N-(3-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-2)

[0115]

[0116] This compound was prepared by a similar preparation method to Example 1. In step 7, N-Boc-1,3-propanediamine was used instead of N-Boc-ethylenediamine, and thus this compound was obtained as a yellow solid. 11H NMR (600 MHz, Chloroform-d) δ 8.73–8.70 (m, 1H), 8.68 (d, J = 1.9 Hz, 2H), 8.38 (d, J = 8.2 Hz, 1H), 7.99 (d, J = 2.1 Hz, 1H), 7.80 (t, J = 5.2 Hz, 1H), 7.56 (d, J = 1.3 Hz, 1H), 7.53 (s, 1H), 7.46 (d, J = 9.4 Hz, 1H), 7.33–7.25 (m, 3H), 7.04 (dd, J = 7.6, 1.0 Hz, 1H), 6.98–6.94 (m, 1H), 6.93–6.89 (m, 1H), 6.83 (dd, J = 8.2, 2.5 Hz, 1H), 4.81 (t, J = 8.1 Hz, 2H), 4.02 (t, J = 6.2 Hz, 2H), 3.97 (s, 2H), 3.56 (q, J = 5.9 Hz, 2H), 3.19 (t, J = 8.1 Hz, 2H), 2.85 (t, J = 5.9 Hz, 2H), 2.75 (t, J = 7.6 Hz, 2H), 2.67 (s, 4H), 2.10–2.02 (m, 2H), 1.82–1.76 (m, 6H). 13 13C NMR (151 MHz, DMSO-d6) δ 163.54, 158.08, 156.91, 153.73, 148.05, 146.74, 143.97, 143.78, 140.70, 140.08, 137.20, 133.72, 132.87, 131.26, 130.03, 129.02, 127.68, 126.50, 123.06, 122.18, 119.91, 119.25, 116.79, 115.38, 113.68, 113.63, 112.85, 65.10, 53.54, 52.94, 51.50, 49.30, 45.62, 36.96, 28.54, 28.02, 27.12, 22.43. ESI-MS m / z: 682.1 [M+H] + 。

[0117] Example 3

[0118] N-(4-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-3)

[0119]

[0120] According to a preparation method similar to that of Example 1, N-Boc-1,4-diaminobutane was used to replace N-Boc-ethylenediamine in Step 7, thereby obtaining this compound, which is a yellow solid. 1 H NMR(600MHz,Chloroform-d)δ8.82(d,J=1.7Hz,1H),8.75(d,J=3.7Hz,2H),8.43(d,J=8.2Hz,1H),8.06(d,J=2.2Hz,1H),7.67(d,J=1.3Hz,1H),7.63(s,1H),7.56(d,J=9.4Hz,1H),7.41(dd,J=9.4,1.8Hz,1H),7.38–7.32(m,2H),7.13–7.09(m,1H),7.05–7.01(m,1H),7.00–6.97(m,2H),6.91(dd,J=8.3,2.6Hz,1H),4.91(t,J=8.1Hz,2H),4.08(t,J=6.2Hz,2H),4.02(s,2H),3.50(q,J=6.5Hz,2H),3.27(t,J=8.1Hz,2H),2.80–2.74(m,4H),2.69(s,4H),2.15–2.07(m,2H),1.86(p,J=3.2Hz,4H),1.77(p,J=7.1Hz,2H),1.67(p,J=6.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ164.52,159.11,157.95,154.80,149.08,147.75,145.04,144.83,141.75,140.77,138.24,134.77,134.01,132.35,131.08,130.06,128.72,127.54,124.11,123.32,120.97,120.37,117.85,116.43,114.73,114.68,113.90,66.11,54.60,53.93,52.49,50.23,48.75,29.08,28.02,27.39,27.12,23.47.ESI-MS m / z:696.5[M+H] + 。

[0121] Example 4

[0122] N-(5-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)pentyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-4)

[0123]

[0124] According to a preparation method similar to that of Example 1, N-Boc-1,5-pentanediamine was used to replace N-Boc-ethylenediamine in Step 7, thereby obtaining this compound, which is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ9.09–9.03(m,1H),8.84(d,J=2.1Hz,1H),8.70(s,1H),8.52(t,J=5.6Hz,1H),8.42(d,J=8.2Hz,1H),8.08(d,J=2.0Hz,1H),8.06–8.03(m,1H),7.65–7.60(m,2H),7.58(d,J=9.4Hz,1H),7.39–7.32(m,2H),7.11(dd,J=7.6,1.0Hz,1H),7.07–7.04(m,1H),7.03(t,J=2.0Hz,1H),6.95(dd,J=8.4,2.5Hz,1H),4.87(t,J=8.1Hz,2H),4.07(t,J=6.4Hz,2H),3.93(s,2H),3.31–3.23(m,4H),2.57–2.52(m,4H),2.46–2.41(m,4H),1.93–1.87(m,2H),1.70–1.63(m,4H),1.58–1.44(m,4H),1.41–1.35(m,2H). 13 C NMR(151MHz,DMSO-d6)δ164.50,159.20,157.96,154.75,149.04,147.81,145.04,144.84,141.74,141.62,138.26,134.76,133.68,132.24,131.05,130.04,128.69,127.53,124.09,123.30,120.88,120.39,117.82,116.43,114.72,114.67,113.86,66.32,54.59,54.08,52.71,50.54,49.11,29.70,29.49,29.08,28.69,24.76,23.56.ESI-MS m / z:710.1[M+H] + 。

[0125] Example 5

[0126] N-(2-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-5)

[0127]

[0128] Prepared by a similar method to that of Example 1. In Step 7, imidazo[1,2-a]pyridine-7-carboxylic acid was used instead of imidazo[1,2-a]pyridine-6-carboxylic acid to obtain this compound as a yellow solid. 1 H NMR(600MHz,Chloroform-d)δ8.78(d,J=2.1Hz,1H),8.74(s,1H),8.44(d,J=8.2Hz,1H),8.14(dd,J=7.1,0.9Hz,1H),8.08(s,1H),8.06(d,J=2.1Hz,1H),7.72(d,J=1.2Hz,1H),7.63(s,1H),7.37–7.32(m,2H),7.31–7.26(m,2H),7.11(dd,J=7.6,1.0Hz,1H),7.04–7.00(m,1H),6.99–6.96(m,1H),6.93–6.88(m,1H),4.89(t,J=8.1Hz,2H),4.08(d,J=6.3Hz,2H),4.05(s,2H),3.63(q,J=5.6Hz,2H),3.24(t,J=8.1Hz,2H),2.99(t,J=5.8Hz,2H),2.73(t,J=7.6Hz,2H),2.64(s,4H),2.13–2.05(m,2H),1.84(p,J=3.1Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ165.24,159.17,157.94,154.76,149.10,147.81,144.83,144.12,141.74,141.47,138.25,135.38,133.80,132.26,131.06,130.46,130.05,127.54,126.99,124.09,120.91,117.83,116.33,114.72,114.51,113.87,110.87,66.27,54.58,54.05,52.66,50.17,48.35,40.04,29.07,28.52,23.53.ESI-MS m / z:668.4[M+H] + 。

[0129] Example 6

[0130] N-(3-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-6)

[0131]

[0132] According to a preparation method similar to that of Example 1, imidazo[1,2-a]pyridine-7-carboxylic acid was used to replace imidazo[1,2-a]pyridine-6-carboxylic acid, and N-Boc-1,3-propanediamine was used to replace N-Boc-ethylenediamine in Step 7, thereby obtaining this compound as a yellow solid. 1 HNMR(600MHz,Chloroform-d)δ8.76(d,J=2.2Hz,1H),8.75(s,1H),8.45(d,J=8.2Hz,1H),8.11(d,J=7.0Hz,1H),8.02(d,J=2.1Hz,1H),7.96(s,1H),7.85(s,1H),7.67(s,1H),7.60(s,1H),7.36(t,J=7.9Hz,2H),7.24(dd,J=7.1,1.7Hz,1H),7.12(d,J=7.6Hz,1H),7.03(d,J=7.6Hz,1H),6.99(t,J=2.1Hz,1H),6.91(dd,J=8.2,2.6Hz,1H),4.87(t,J=8.1Hz,2H),4.09(t,J=6.2Hz,2H),4.05(s,2H),3.64(q,J=5.8Hz,2H),3.26(t,J=8.1Hz,2H),2.91(t,J=5.9Hz,2H),2.82–2.77(m,2H),2.71(s,4H),2.16–2.09(m,2H),1.90–1.83(m,6H). 1313C NMR(151MHz,DMSO-d6)δ165.07,159.18,157.94,154.75,149.06,147.79,144.84,144.11,141.76,141.34,138.25,135.36,133.82,132.28,131.06,130.48,130.05,127.54,127.00,124.09,120.92,117.84,116.20,114.73,114.50,113.87,110.79,66.26,54.58,54.04,52.65,50.42,46.69,38.09,29.67,29.07,28.49,23.53.ESI-MS m / z:682.5[M+H] + 。

[0133] Example 7

[0134] N-(4-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-7)

[0135]

[0136] Prepared by a method similar to that of Example 1. In step 7, imidazo[1,2-a]pyridine-7-carboxylic acid was used instead of imidazo[1,2-a]pyridine-6-carboxylic acid, and N-Boc-1,4-butanediamine was used instead of N-Boc-ethylenediamine to obtain this compound as a yellow solid. 1HNMR(600MHz,Chloroform-d)δ8.75(s,2H),8.43(d,J=8.1Hz,1H),8.12(d,J=7.0Hz,1H),8.03(d,J=2.2Hz,1H),8.00(s,1H),7.70(s,1H),7.61(s,1H),7.38–7.32(m,2H),7.31–7.27(m,1H),7.11(d,J=7.5Hz,1H),7.03(d,J=7.6Hz,1H),7.00–6.97(m,1H),6.91(dd,J=8.3,2.6Hz,1H),4.91(t,J=8.1Hz,2H),4.09(t,J=6.3Hz,2H),4.02(s,2H),3.51(q,J=6.5Hz,2H),3.27(t,J=8.1Hz,2H),2.77(t,J=6.7Hz,2H),2.73(t,J=7.5Hz,2H),2.64(s,4H),2.09(p,J=6.6Hz,2H),1.84(p,J=3.2Hz,4H),1.78(p,J=7.1Hz,2H),1.67(p,J=6.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ164.97,159.16,157.95,154.77,149.05,147.79,144.84,144.13,141.75,141.23,138.24,135.37,133.83,132.29,131.06,130.54,130.05,127.54,127.00,124.09,120.92,117.83,116.21,114.72,114.50,113.88,110.84,66.23,54.59,54.02,52.62,50.40,48.86,29.08,28.40,27.39,27.33,23.52.ESI-MS m / z:696.3[M+H] + 。

[0137] Example 8

[0138] N-(5-(((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)pentyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-8)

[0139]

[0140] Prepared by a similar method to Example 1, in step 7, imidazo[1,2-a]pyridine-7-carboxylic acid was used to replace imidazo[1,2-a]pyridine-6-carboxylic acid, and N-Boc-1,5-pentanediamine was used to replace N-Boc-ethylenediamine to obtain this compound, which is a yellow solid. ESI-MS m / z: 710.8 [M+H] + 。

[0141] Example 9

[0142] 3-(1H-Imidazol-1-yl)-N-((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)propan-1-amine (Compound I-9)

[0143]

[0144] Dissolve 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde (120 mg, 0.25 mmol) and 3-(1H-imidazol-1-yl)propan-1-amine (47 mg, 0.38 mmol) in 4 mL of methanol and 2 mL of dichloromethane solution, add a drop of glacial acetic acid, react at room temperature for 30 min, then add NaBH3CN (39.3 mg, 0.63 mmol) and continue to react at room temperature for 2 h. After the reaction is completed, pour the reaction solution into 20 mL of water, extract with dichloromethane (15 mL×3), combine the organic phases, concentrate under reduced pressure, and purify the crude product by column chromatography to obtain 85 mg of yellow solid, with a yield of 50.87%. 11H NMR (600 MHz, DMSO-d6) δ 8.86 (d, J = 2.1 Hz, 1H), 8.72 (s, 1H), 8.43 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 2.1 Hz, 1H), 7.59 (s, 1H), 7.40–7.33 (m, 2H), 7.15 (d, J = 1.3 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 7.09–7.06 (m, 1H), 7.05–7.02 (m, 1H), 6.96 (dd, J = 8.3, 2.5 Hz, 1H), 6.87–6.84 (m, 1H), 4.89 (t, J = 8.1 Hz, 2H), 4.09 (t, J = 6.3 Hz, 2H), 4.04 (t, J = 7.0 Hz, 2H), 3.93 (s, 2H), 3.27 (t, J = 8.1 Hz, 2H), 2.68 (s, 2H), 2.60 (s, 4H), 2.50–2.47 (m, 2H), 1.95 (p, J = 6.8 Hz, 2H), 1.89 (p, J = 6.9 Hz, 2H), 1.74–1.71 (m, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 159.09, 158.03, 154.84, 149.17, 147.81, 144.85, 141.77, 141.23, 138.27, 137.70, 133.92, 132.33, 131.13, 130.11, 128.75, 127.59, 124.15, 121.03, 119.77, 117.86, 114.74, 113.95, 66.04, 54.65, 53.93, 52.43, 50.30, 45.76, 44.37, 31.23, 29.11, 27.79, 23.44. ESI-MS m / z: 589.2 [M+H] + 。

[0145] Replace 1-(3-chloropropyl)pyrrolidine in Step 5 of Example 1 with 3-chloro-N,N-diethylpropan-1-amine, and then prepare the target compounds in Examples 10 to 17 according to a preparation procedure similar to that of Examples 1 to 8.

[0146] Example 10

[0147] N-(2-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-10)

[0148]

[0149] 1 1H NMR (600 MHz, DMSO-d6) δ 9.12–9.08 (m, 1H), 8.88 (d, J = 2.1 Hz, 1H), 8.70 (s, 1H), 8.57 (t, J = 5.6 Hz, 1H), 8.42 (d, J = 8.1 Hz, 1H), 8.12 (d, J = 2.1 Hz, 1H), 8.07–8.03 (m, 1H), 7.66–7.62 (m, 2H), 7.59 (d, J = 9.4 Hz, 1H), 7.41–7.37 (m, 1H), 7.36–7.32 (m, 1H), 7.11 (dd, J = 7.6, 1.0 Hz, 1H), 7.09–7.05 (m, 1H), 7.05–7.01 (m, 1H), 6.98–6.94 (m, 1H), 4.85 (t, J = 8.2 Hz, 2H), 4.08 (t, J = 6.3 Hz, 2H), 4.00 (s, 2H), 3.44 (q, J = 6.3 Hz, 2H), 3.25 (t, J = 8.2 Hz, 2H), 2.76 (t, J = 6.5 Hz, 2H), 2.69–2.55 (m, 6H), 1.91–1.86 (m, 2H), 0.99 (t, J = 7.1 Hz, 6H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.78, 159.15, 157.99, 154.79, 149.14, 147.82, 145.05, 144.84, 141.74, 141.41, 138.26, 134.79, 133.86, 132.29, 131.08, 130.08, 128.81, 127.56, 124.11, 123.37, 120.95, 120.32, 117.85, 116.42, 114.75, 114.69, 113.89, 66.11, 54.59, 50.12, 49.02, 48.35, 46.85, 29.48, 29.07, 26.45, 11.65. ESI-MS m / z: 670.8 [M+H] + 。

[0150] Example 11

[0151] N-(3-(((4-(4-(3-(3-(diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-11)

[0152]

[0153] 1 1H NMR (600 MHz, Chloroform-d) δ 8.79 (s, 1H), 8.76 (s, 2H), 8.46 (d, J = 8.2 Hz, 1H), 8.07 (d, J = 2.2 Hz, 1H), 7.85 (d, J = 5.2 Hz, 1H), 7.64 (s, 1H), 7.61 (s, 1H), 7.54 (d, J = 9.4 Hz, 1H), 7.40–7.33 (m, 3H), 7.12 (d, J = 7.6 Hz, 1H), 7.04 (d, J = 7.5 Hz, 1H), 6.99–6.96 (m, 1H), 6.90 (dd, J = 8.3, 2.6 Hz, 1H), 4.88 (t, J = 8.1 Hz, 2H), 4.08 (t, J = 6.0 Hz, 2H), 4.04 (s, 2H), 3.63 (q, J = 5.7 Hz, 2H), 3.27 (t, J = 8.1 Hz, 2H), 2.93 (t, J = 5.8 Hz, 2H), 2.82 (t, J = 7.7 Hz, 2H), 2.74 (q, J = 7.1 Hz, 4H), 2.12–2.03 (m, 2H), 1.91–1.83 (m, 2H), 1.14 (t, J = 7.2 Hz, 6H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.61, 159.08, 157.97, 154.83, 149.17, 147.74, 145.03, 144.82, 141.76, 140.28, 138.25, 134.78, 134.29, 132.42, 131.11, 130.10, 128.79, 127.57, 124.14, 123.27, 121.02, 120.27, 117.87, 116.42, 114.76, 114.69, 113.92, 65.96, 54.60, 50.04, 48.83, 46.77, 46.50, 37.87, 29.25, 29.08, 25.82, 11.04. ESI-MS m / z: 684.7 [M+H] + 。

[0154] Example 12

[0155] N-(4-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-12)

[0156]

[0157] 11H NMR (600 MHz, Chloroform-d) δ 8.82 (s, 1H), 8.75 (d, J = 1.7 Hz, 2H), 8.43 (t, J = 7.4 Hz, 1H), 8.07 (d, J = 2.2 Hz, 1H), 7.66 (s, 1H), 7.63 (s, 1H), 7.56 (d, J = 9.4 Hz, 1H), 7.42 (dd, J = 9.5, 1.8 Hz, 1H), 7.35 (dd, J = 10.6, 5.3 Hz, 2H), 7.11 (d, J = 8.0 Hz, 1H), 7.05–6.95 (m, 3H), 6.90 (dd, J = 8.6, 2.9 Hz, 1H), 4.91 (q, J = 7.2 Hz, 2H), 4.07 (dd, J = 8.0, 4.3 Hz, 2H), 4.01 (d, J = 5.2 Hz, 2H), 3.50 (q, J = 6.5 Hz, 2H), 3.29–3.21 (m, 2H), 2.76 (t, J = 6.9 Hz, 2H), 2.69 (q, J = 7.3 Hz, 4H), 2.03 (d, J = 6.7 Hz, 4H), 1.80–1.73 (m, 2H), 1.66 (dd, J = 15.0, 7.9 Hz, 2H), 1.12 (t, J = 7.0 Hz, 6H). ESI-MS m / z: 698.5 [M+H] + 。

[0158] Example 13

[0159] N-(5-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)pentyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-13)

[0160]

[0161] ESI-MS m / z: 712.5 [M+H] + 。

[0162] Example 14

[0163] N-(2-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-14)

[0164]

[0165] ESI-MS m / z: 670.7 [M+H] + 。

[0166] Example 15

[0167] N-(3-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-15)

[0168]

[0169] 1 H NMR(600MHz,Chloroform-d)δ8.77–8.72(m,2H),8.45(d,J=8.2Hz,1H),8.11(d,J=7.0Hz,1H),8.03(s,1H),7.97(s,1H),7.88(d,J=6.5Hz,1H),7.67(s,1H),7.60(s,1H),7.38–7.33(m,2H),7.24(d,J=7.4Hz,1H),7.12(d,J=7.6Hz,1H),7.03(d,J=7.5Hz,1H),6.99(s,1H),6.91(dd,J=8.0,2.5Hz,1H),4.87(t,J=8.1Hz,2H),4.09–4.03(m,4H),3.63(q,J=5.8Hz,2H),3.26(t,J=8.1Hz,2H),2.90(t,J=6.0Hz,2H),2.72(t,J=7.5Hz,2H),2.64(q,J=7.2Hz,4H),2.04–1.98(m,2H),1.90–1.85(m,2H),1.09(t,J=7.2Hz,6H). 13 C NMR(151MHz,DMSO-d6)δ165.07,159.19,157.96,154.76,149.08,147.79,144.84,144.11,141.75,141.25,138.26,135.36,133.87,132.30,131.07,130.47,130.07,127.55,127.00,124.09,120.92,117.84,116.20,114.75,114.50,113.86,110.79,66.19,54.58,50.39,49.09,46.86,46.67,38.07,29.63,29.08,26.74,11.92.ESI-MS m / z:684.4[M+H] + 。

[0170] Example 16

[0171] N-(4-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-16)

[0172]

[0173] 1 H NMR (600 MHz, Chloroform-d) δ 8.75 (d, J = 2.5 Hz, 2H), 8.43 (d, J = 8.1 Hz, 1H), 8.13 (d, J = 7.1 Hz, 1H), 8.04–7.98 (m, 2H), 7.70 (s, 1H), 7.61 (s, 1H), 7.38–7.32 (m, 2H), 7.29 (d, J = 7.0 Hz, 1H), 7.11 (d, J = 7.5 Hz, 1H), 7.02 (d, J = 7.6 Hz, 1H), 6.98 (s, 1H), 6.91 (dd, J = 8.3, 2.6 Hz, 1H), 4.91 (t, J = 8.0 Hz, 2H), 4.06 (t, J = 6.2 Hz, 2H), 4.02 (s, 2H), 3.51 (q, J = 6.5 Hz, 2H), 3.27 (t, J = 8.0 Hz, 2H), 2.77 (t, J = 6.7 Hz, 2H), 2.69 (t, J = 7.5 Hz, 2H), 2.61 (q, J = 7.2 Hz, 4H), 2.01–1.96 (m, 2H), 1.80–1.75 (m, 2H), 1.70–1.65 (m, 2H), 1.07 (t, J = 7.2 Hz, 6H). ESI-MS m / z: 698.5 [M+H] + 。

[0174] Example 17

[0175] N-(5-(((4-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)pentyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-17)

[0176]

[0177] ESI-MS m / z: 712.5 [M+H] + 。

[0178] Example 18

[0179] (E)-2-Cyano-1-(pyridin-4-yl)-3-((4-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)guanidine (Compound I-18)

[0180]

[0181] Step 1: (4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanol

[0182]

[0183] Under an ice bath, NaBH4 (0.095 g, 2.5 mmol) was slowly added to 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde (1.0 g, 2.09 mmol) in a mixed solvent of methanol (20 mL) and dichloromethane (8 mL). After addition, the reaction was carried out at room temperature for 10 min. After the reaction was completed, 0.2 mL of saturated ammonium chloride aqueous solution was added to quench the reaction. The reaction solution was poured into 40 mL of water and extracted with dichloromethane (40 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was evaporated to dryness under reduced pressure to obtain 0.98 g of a yellow solid with a yield of 97.6%. 1 1H NMR (600 MHz, Chloroform-d) δ 8.69 (d, J = 2.1 Hz, 1H), 8.66 (s, 1H), 8.35 (d, J = 8.2 Hz, 1H), 8.05 (dd, J = 2.3, 1.1 Hz, 1H), 7.28–7.24 (m, 2H), 7.03 (dd, J = 7.6, 1.0 Hz, 1H), 6.94–6.91 (m, 1H), 6.90–6.88 (m, 1H), 6.84–6.81 (m, 1H), 4.85 (s, 2H), 4.82 (t, J = 8.1 Hz, 2H), 4.00 (t, J = 6.3 Hz, 2H), 3.72 (s, 1H), 3.17 (t, J = 8.1 Hz, 2H), 2.62 (t, J = 7.6 Hz, 2H), 2.52 (d, J = 6.0 Hz, 4H), 1.99 (q, J = 7.0 Hz, 2H), 1.79–1.71 (m, 4H). ESI-MS m / z: 482.3 [M+H] + 。

[0184] Step 2: 7-(Chloromethyl)-4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine

[0185]

[0186] At room temperature, (4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanol (0.98 g, 2.03 mmol) was slowly added to 5 mL of thionyl chloride, and the reaction was carried out at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated, and the residue was added to 30 mL of saturated NaHCO3 solution. It was extracted with dichloromethane (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was evaporated to dryness to obtain 0.97 g of a yellow solid, with a yield of 95.3%. 1 1H NMR (600 MHz, Chloroform-d) δ 8.76 (d, J = 2.2 Hz, 1H), 8.72 (s, 1H), 8.40 (d, J = 8.2 Hz, 1H), 8.06 (d, J = 2.3 Hz, 1H), 7.32–7.26 (m, 2H), 7.05 (dd, J = 7.6, 1.0 Hz, 1H), 7.00–6.95 (m, 1H), 6.90–6.88 (m, 1H), 6.85–6.80 (m, 1H), 4.87 (t, J = 8.1 Hz, 2H), 4.69 (s, 2H), 4.03 (t, J = 5.9 Hz, 2H), 3.21 (t, J = 8.1 Hz, 2H), 2.98–2.82 (m, 6H), 2.21–2.15 (m, 2H), 1.92 (t, J = 3.4 Hz, 4H). ESI-MS m / z: 500.1 [M+H] + 。

[0187] Step 3: 2-((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)isoindoline-1,3-dione

[0188]

[0189] The intermediate 7-(chloromethyl)-4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine (0.95 g, 1.9 mmol) and potassium phthalimide (0.53 g, 2.85 mmol) were added to 10 mL of DMF, and the reaction was carried out at 40 °C for 4 h. After the reaction was completed, the reaction solution was poured into 150 mL of water, and a solid was precipitated. It was filtered by suction and dried to obtain 0.95 g of a yellow solid, with a yield of 81.9%. 11H NMR (600 MHz, Chloroform-d) δ 8.80 (d, J = 2.2 Hz, 1H), 8.68 (s, 1H), 8.38 (d, J = 8.2 Hz, 1H), 8.02 (d, J = 2.2 Hz, 1H), 7.84–7.79 (m, 2H), 7.72–7.66 (m, 2H), 7.30–7.25 (m, 2H), 7.03 (d, J = 7.2 Hz, 1H), 7.00–6.93 (m, 1H), 6.89 (t, J = 1.9 Hz, 1H), 6.82 (dd, J = 8.2, 2.6 Hz, 1H), 4.99 (s, 2H), 4.85 (t, J = 8.1 Hz, 2H), 4.02 (t, J = 6.1 Hz, 2H), 3.19 (t, J = 8.1 Hz, 2H), 2.79 (d, J = 32.5 Hz, 6H), 2.16–2.07 (m, 2H), 1.90–1.81 (m, 4H). ESI-MS m / z: 611.4 [M+H] + 。

[0190] Step 4: (4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine

[0191]

[0192] Dissolve 2-((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)isoindoline-1,3-dione (1.0 g, 1.64 mmol) in 20 mL of ethanol, add hydrazine hydrate (0.16 g, 3.27 mmol) dropwise, and react at 80 °C for 6 h. After the reaction is completed, pour the reaction solution into 50 mL of water, extract with dichloromethane (40 mL × 3), combine the organic phases, wash with saturated brine (80 mL), dry over anhydrous sodium sulfate, filter by suction, evaporate the filtrate to dryness, and purify the crude product by column chromatography to obtain 0.62 g of a yellow solid with a yield of 78.8%. 11H NMR (600 MHz, Chloroform-d) δ 8.80–8.77 (m, 2H), 8.43 (d, J = 8.2 Hz, 1H), 8.06 (q, J = 1.1 Hz, 1H), 7.37–7.32 (m, 2H), 7.11 (d, J = 7.6 Hz, 1H), 7.03 (dd, J = 7.7, 1.5 Hz, 1H), 6.99–6.97 (m, 1H), 6.90 (dd, J = 8.3, 2.5 Hz, 1H), 4.93 (t, J = 8.1 Hz, 2H), 4.12 (s, 2H), 4.09 (t, J = 6.2 Hz, 2H), 3.28 (t, J = 8.1 Hz, 2H), 2.81 (t, J = 7.6 Hz, 2H), 2.74 (s, 4H), 2.14 (p, J = 6.6 Hz, 2H), 1.92–1.85 (m, 4H). ESI-MS m / z: 481.6 [M+H] + 。

[0193] Step 5: (E)-2-Cyano-1-(pyridin-4-yl)-3-((4-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)guanidine (Compound I-18)

[0194]

[0195] (4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine (100 mg, 0.21 mmol), methyl (Z)-N'-cyano-N-(pyridin-4-yl)carbamimidothioate (48 mg, 0.25 mmol), 4-dimethylaminopyridine (2.54 mg, 0.021 mmol) and triethylamine (31.6 mg, 0.31 mmol) were successively added to 4 mL of pyridine, and the reaction was carried out at 70 °C for 5 h. After the reaction was completed, the reaction solution was evaporated to dryness, and the crude product was separated and purified by column chromatography to obtain 60 mg of a yellow solid with a yield of 46.2%. 11H NMR (600 MHz, DMSO-d6) δ 9.78 (bs, 1H), 8.87 (d, J = 2.2 Hz, 1H), 8.73 (s, 1H), 8.52 (s, 1H), 8.44 (d, J = 8.2 Hz, 1H), 8.40 (d, J = 5.4 Hz, 2H), 8.07 (d, J = 2.1 Hz, 1H), 7.41–7.32 (m, 2H), 7.22 (s, 2H), 7.12 (d, J = 7.6 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 7.04–7.01 (m, 1H), 6.96 (dd, J = 8.3, 2.5 Hz, 1H), 4.90 (t, J = 8.1 Hz, 2H), 4.73 (s, 2H), 4.08 (t, J = 6.4 Hz, 2H), 3.27 (t, J = 8.1 Hz, 2H), 2.57 (t, J = 7.2 Hz, 2H), 2.49–2.44 (m, 4H), 1.91 (p, J = 6.7 Hz, 2H), 1.70–1.67 (m, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 159.19, 158.40, 157.94, 155.04, 150.21, 148.17, 147.65, 146.70, 144.76, 141.72, 138.82, 138.30, 133.61, 132.62, 131.14, 130.07, 127.58, 124.25, 120.91, 117.92, 116.71, 115.61, 114.72, 113.90, 66.29, 54.64, 54.08, 52.68, 43.14, 29.10, 28.58, 23.55. ESI-MS m / z: 625.2 [M+H] + 。

[0196] Example 19

[0197] N-((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-19)

[0198]

[0199] At room temperature, (4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine (80 mg, 0.17 mmol), imidazo[1,2-a]pyridine-6-carboxylic acid (32.4 mg, 0.20 mmol), and HATU (82.3 mg, 0.22 mmol) were successively added to 4 mL of DMF. After stirring for 10 min, DIPEA (64.5 mg, 0.50 mmol) was added, and the reaction was continued at room temperature for 30 min. After the reaction was completed, the reaction solution was poured into 20 mL of water, and extracted with ethyl acetate (15 mL×3). The organic layers were combined, washed twice with water, once with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, and the obtained filtrate was concentrated under reduced pressure. The residue was purified by column chromatography to obtain 82 mg of a yellow solid, with a yield of 78.9%. 1 H NMR(600MHz,DMSO-d6)δ9.38(t,J=5.8Hz,1H),9.23–9.20(m,1H),8.91(d,J=2.2Hz,1H),8.72(s,1H),8.44(d,J=8.2Hz,1H),8.11–8.07(m,2H),7.71(dd,J=9.5,1.8Hz,1H),7.68–7.62(m,2H),7.41–7.32(m,2H),7.12(d,J=7.5Hz,1H),7.06(d,J=8.0Hz,1H),7.04–7.01(m,1H),6.95(dd,J=8.3,2.5Hz,1H),4.89(t,J=8.1Hz,2H),4.76(d,J=5.7Hz,2H),4.08(t,J=6.3Hz,2H),3.27(t,J=8.1Hz,2H),2.60(d,J=54.2Hz,6H),1.93(p,J=6.8Hz,2H),1.75–1.65(m,4H). 13 C NMR(151MHz,DMSO-d6)δ165.16,159.14,157.93,154.97,148.28,147.71,145.10,144.77,141.73,139.80,138.28,134.94,133.49,132.48,131.12,130.06,129.22,127.57,124.20,123.21,120.94,119.77,117.91,116.65,114.84,114.73,113.89,66.18,54.61,54.02,52.59,40.94,29.08,28.29,23.50.ESI-MS m / z:625.4[M+H] +。

[0200] Example 20

[0201] (E)-3-(Pyridin-3-yl)-N-((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-20)

[0202]

[0203] Prepared by a similar method to Example 19, replacing imidazo[1,2-a]pyridine-6-carboxylic acid with (E)-3-(pyridin-3-yl)acrylic acid, the compound is a yellow solid. 1 H NMR (600 MHz, DMSO-d6) δ 8.98 (t, J = 5.9 Hz, 1H), 8.84 (d, J = 2.1 Hz, 1H), 8.80 (d, J = 2.2 Hz, 1H), 8.71 (s, 1H), 8.57 (dd, J = 4.7, 1.6 Hz, 1H), 8.44 (d, J = 8.2 Hz, 1H), 8.05–8.00 (m, 2H), 7.56 (d, J = 15.9 Hz, 1H), 7.45 (dd, J = 8.0, 4.7 Hz, 1H), 7.39–7.30 (m, 2H), 7.11 (d, J = 7.6 Hz, 1H), 7.06–6.99 (m, 2H), 6.94 (dd, J = 8.3, 2.5 Hz, 1H), 6.87 (d, J = 15.9 Hz, 1H), 4.86 (t, J = 8.1 Hz, 2H), 4.67 (d, J = 5.8 Hz, 2H), 4.06 (t, J = 6.4 Hz, 2H), 3.25 (t, J = 8.1 Hz, 2H), 2.55–2.51 (m, 2H), 2.44–2.38 (m, 4H), 1.89 (p, J = 6.8 Hz, 2H), 1.70–1.61 (m, 4H). 1313C NMR (151 MHz, DMSO-d6) δ 165.45, 159.21, 157.94, 154.99, 150.72, 149.70, 148.26, 147.72, 144.77, 141.72, 139.83, 138.30, 136.62, 134.51, 133.33, 132.45, 131.12, 131.04, 130.05, 127.56, 124.43, 124.21, 124.07, 120.88, 117.90, 114.72, 113.89, 66.34, 54.61, 54.09, 52.72, 40.49, 29.08, 28.73, 23.57. ESI-MS m / z: 612.1 [M+H] + 。

[0204] Example 21

[0205] (E)-2-Cyano-1-(pyridin-4-yl)-3-(2-(((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)guanidine (Compound I-21)

[0206]

[0207] Step 1: tert-Butyl (2-(((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)carbamate

[0208]

[0209] To a mixed solution of methanol (4 mL) and dichloromethane (2 mL) containing 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde (0.12 g, 0.25 mmol) and N-Boc-ethylenediamine (60.1 mg, 0.38 mmol) was added 1 drop of glacial acetic acid, and the mixture was stirred at room temperature for 30 min. Then, NaBH3CN (39.3 mg, 0.63 mmol) was added, and the reaction was continued at room temperature for 1 h. After completion of the reaction, the reaction solution was poured into 20 mL of water, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was evaporated to dryness. The crude product was purified by column chromatography to obtain 110 mg of a yellow solid with a yield of 70.5%. 1HNMR(600MHz, DMSO-d6) δ 8.85 (d, J = 2.1 Hz, 1H), 8.72 (s, 1H), 8.42 (d, J = 8.2 Hz, 1H), 8.09 (d, J = 2.0 Hz, 1H), 7.40–7.33 (m, 2H), 7.12 (d, J = 7.6 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 7.03 (t, J = 2.1 Hz, 1H), 6.95 (dd, J = 8.3, 2.6 Hz, 1H), 6.78 (t, J = 5.7 Hz, 1H), 4.89 (t, J = 8.1 Hz, 2H), 4.08 (t, J = 6.4 Hz, 2H), 3.93 (s, 2H), 3.27 (t, J = 8.2 Hz, 2H), 3.06 (q, J = 6.3 Hz, 2H), 2.60–2.55 (m, 4H), 2.46 (d, J = 5.9 Hz, 4H), 1.94–1.88 (m, 2H), 1.71–1.66 (m, 4H), 1.37 (s, 9H). ESI-MS m / z: 624.4 [M+H] + 。

[0210] Step 2: N 1 -((4-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)ethane-1,2-diamine

[0211]

[0212] To a solution of tert-butyl (2-(((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)carbamate (100 mg, 0.16 mmol) in methanol (4 mL) was added 1 mL of dioxane hydrochloride solution, and the reaction was carried out at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated, 15 mL of saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was evaporated to dryness to obtain 80 mg of a yellow solid with a yield of 95.3%. ESI-MS m / z: 524.6 [M+H] + 。

[0213] Step 3: (E)-2-Cyano-1-(pyridin-4-yl)-3-(2-(((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)ethyl)guanidine (Compound I-21)

[0214]

[0215] Prepared by a similar preparation method as in step 5 of Example 18 using N 1 -((4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)ethane-1,2-diamine as the raw material to obtain the target compound. ESI-MS m / z: 668.2 [M+H] + 。

[0216] Example 22

[0217] (S)-N-(3-(((4-(4-(3-(3-(3-hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-22)

[0218]

[0219] Step 1: 4-(4-(3-(3-chloropropoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine

[0220]

[0221] The intermediate 3-(1-(7-bromopyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol (5 g, 13.7 mmol) obtained in step 3 of Example 1, 1-bromo-3-chloropropane (2.36 g, 15.0 mmol), and potassium carbonate (4.71 g, 34.1 mmol) were successively added to 50 mL of DMF, and the mixture was stirred at 70 °C for 4 h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with 100 mL of ethyl acetate, and then filtered through diatomaceous earth. The filter cake was washed with ethyl acetate several times. The filtrate was poured into 300 mL of water, and extracted with ethyl acetate (200 mL×3). The organic phase was washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain 5.3 g of a yellow solid, with a yield of 87.7%. 11H NMR (600 MHz, Chloroform-d) δ 8.89 (d, J = 2.2 Hz, 1H), 8.77 (d, J = 1.2 Hz, 1H), 8.44 (d, J = 8.1 Hz, 1H), 8.09 (d, J = 2.2 Hz, 1H), 7.38–7.34 (m, 2H), 7.12 (dd, J = 7.5, 1.0 Hz, 1H), 7.07–7.03 (m, 1H), 7.00 (dd, J = 2.6, 1.6 Hz, 1H), 6.94–6.91 (m, 1H), 6.89 (dd, J = 17.7, 11.0 Hz, 1H), 6.09 (d, J = 17.6 Hz, 1H), 5.61 (d, J = 11.0 Hz, 1H), 4.94 (dd, J = 9.5, 6.6 Hz, 2H), 4.17 (t, J = 5.9 Hz, 2H), 3.78 (t, J = 6.3 Hz, 2H), 3.31–3.27 (m, 2H), 2.27 (p, J = 6.1 Hz, 2H). ESI-MS m / z: 443.6 [M+H] + 。

[0222] Step 2: (S)-1-(3-(3-(1-(7-Vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenoxy)propyl)pyrrolidin-3-ol

[0223]

[0224] The intermediate 4-(4-(3-(3-chloropropoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine (2.5 g, 5.64 mmol), (S)-3-hydroxypyrrolidine hydrochloride (0.84 g, 6.77 mmol), potassium carbonate (2.34 g, 16.9 mmol) and potassium iodide (0.094 g, 0.56 mmol) were successively added to 20 mL of DMF, and the reaction was carried out at 70 °C for 8 h. After the reaction was completed, the reaction solution was poured into 200 mL of water, and extracted with ethyl acetate (200 mL × 3). The organic phase was washed twice with water, once with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, the filtrate was evaporated to dryness, and the crude product was separated and purified by column chromatography to obtain 2.05 g of a yellow solid with a yield of 73.6%. 11H NMR (400 MHz, Chloroform-d) δ 8.81 (d, J = 2.2 Hz, 1H), 8.70 (s, 1H), 8.38–8.33 (m, 1H), 7.99 (d, J = 2.2 Hz, 1H), 7.32–7.24 (m, 2H), 7.06–7.01 (m, 1H), 6.98–6.94 (m, 1H), 6.92 (dd, J = 2.6, 1.5 Hz, 1H), 6.86–6.77 (m, 2H), 6.01 (d, J = 17.7 Hz, 1H), 5.53 (d, J = 11.0 Hz, 1H), 4.86 (t, J = 8.1 Hz, 2H), 4.29 (t, J = 6.4 Hz, 1H), 4.01 (t, J = 6.3 Hz, 2H), 3.21 (t, J = 8.1 Hz, 2H), 2.93–2.86 (m, 1H), 2.70 (d, J = 9.7 Hz, 1H), 2.66–2.59 (m, 2H), 2.52 (dd, J = 10.1, 5.1 Hz, 1H), 2.34–2.24 (m, 1H), 2.23–2.07 (m, 2H), 2.02–1.93 (m, 2H), 1.76–1.67 (m, 1H). ESI-MS m / z: 494.3 [M+H] + 。

[0225] Step 3: (S)-4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0226]

[0227] According to a preparation method similar to that in Step 6 of Example 1, replace 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine with (S)-1-(3-(3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenoxy)propyl)pyrrolidin-3-ol to obtain this intermediate, which is a yellow solid with a yield of 70.7%. 11H NMR (600 MHz, Chloroform-d) δ 10.30 (s, 1H), 9.24 (d, J = 2.0 Hz, 1H), 8.86 (s, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.51 (d, J = 8.2 Hz, 1H), 7.40–7.34 (m, 2H), 7.16 (d, J = 7.6 Hz, 1H), 7.03 (d, J = 7.5 Hz, 1H), 7.00–6.97 (m, 1H), 6.92 (dd, J = 8.2, 2.6 Hz, 1H), 4.97 (t, J = 8.0 Hz, 2H), 4.36 (t, J = 7.6 Hz, 1H), 4.09 (t, J = 6.3 Hz, 2H), 3.31 (t, J = 8.0 Hz, 2H), 2.98–2.93 (m, 1H), 2.76 (d, J = 10.0 Hz, 1H), 2.69 (t, J = 7.3 Hz, 2H), 2.56 (dd, J = 10.1, 5.2 Hz, 1H), 2.37–2.32 (m, 1H), 2.24–2.17 (m, 2H), 2.04 (p, J = 6.6 Hz, 1H), 1.95–1.85 (m, 1H), 1.80–1.74 (m, 1H). ESI-MS m / z: 496.2 [M+H] + 。

[0228] Step 4: (S)-N-(3-(((4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-22)

[0229]

[0230] According to a preparation method similar to that of Example 2, (S)-4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde was used to replace 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde, thereby obtaining this compound as a yellow solid. 11H NMR (600 MHz, DMSO-d6) δ 9.09–9.06 (m, 1H), 8.86 (d, J = 2.1 Hz, 1H), 8.69 (s, 1H), 8.60 (t, J = 5.5 Hz, 1H), 8.42 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 2.1 Hz, 1H), 8.04 (s, 1H), 7.63–7.59 (m, 2H), 7.57 (d, J = 9.4 Hz, 1H), 7.40–7.32 (m, 2H), 7.11 (dd, J = 7.7, 1.0 Hz, 1H), 7.06 (dd, J = 7.6, 1.3 Hz, 1H), 7.04–7.02 (m, 1H), 6.96 (dd, J = 8.3, 2.5 Hz, 1H), 4.85 (t, J = 8.2 Hz, 2H), 4.71 (s, 1H), 4.21–4.15 (m, 1H), 4.07 (t, J = 6.4 Hz, 2H), 3.96 (s, 2H), 3.37 (q, J = 6.5 Hz, 2H), 3.25 (t, J = 8.1 Hz, 2H), 2.74–2.70 (m, 1H), 2.65–2.53 (m, 5H), 2.48–2.42 (m, 1H), 2.35 (dd, J = 9.7, 3.7 Hz, 1H), 2.02–1.94 (m, 1H), 1.89 (p, J = 6.8 Hz, 2H), 1.75 (p, J = 6.9 Hz, 2H), 1.58–1.50 (m, 1H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.59, 159.18, 157.96, 154.77, 149.08, 147.80, 145.03, 144.83, 141.75, 141.38, 138.26, 134.77, 133.81, 132.28, 131.07, 130.06, 128.73, 127.54, 124.11, 123.24, 120.91, 120.32, 117.84, 116.43, 114.74, 114.69, 113.87, 69.80, 66.26, 63.18, 54.59, 52.99, 52.89, 50.44, 46.72, 38.05, 34.84, 29.69, 29.07, 28.38. ESI-MS m / z: 698.2 [M+H] + 。

[0231] Example 23

[0232] (S)-N-(4-(((4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-23)

[0233]

[0234] Prepared by a similar preparation method as in Example 3, using (S)-4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material through reductive amination reaction. ESI-MS m / z: 712.5 [M+H] + 。

[0235] Prepared by a similar preparation method as in Examples 22 and 23, using (R)-3-Hydroxypyrrolidine hydrochloride to replace (S)-3-Hydroxypyrrolidine hydrochloride in Step 2 of Example 22 to obtain the compounds described in Examples 24 and 25.

[0236] Example 24

[0237] (R)-N-(3-(((4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-24)

[0238]

[0239] 11H NMR (600 MHz, DMSO-d6) δ 9.13–9.04 (m, 1H), 8.85 (d, J = 2.1 Hz, 1H), 8.69 (s, 1H), 8.59 (t, J = 5.5 Hz, 1H), 8.42 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 2.0 Hz, 1H), 8.04 (s, 1H), 7.63 (d, J = 1.2 Hz, 1H), 7.60 (dd, J = 9.5, 1.8 Hz, 1H), 7.57 (d, J = 9.4 Hz, 1H), 7.41–7.35 (m, 1H), 7.37–7.31 (m, 1H), 7.11 (d, J = 7.6 Hz, 1H), 7.08–7.04 (m, 1H), 7.03 (t, J = 2.0 Hz, 1H), 6.96 (dd, J = 8.3, 2.5 Hz, 1H), 4.85 (t, J = 8.1 Hz, 2H), 4.69 (d, J = 4.5 Hz, 1H), 4.21–4.15 (m, 1H), 4.07 (t, J = 6.4 Hz, 2H), 3.95 (s, 2H), 3.37 (q, J = 6.6 Hz, 2H), 3.25 (t, J = 8.1 Hz, 2H), 2.71 (dd, J = 9.6, 6.2 Hz, 1H), 2.63 (t, J = 6.8 Hz, 2H), 2.62–2.50 (m, 3H), 2.46–2.41 (m, 1H), 2.36–2.31 (m, 1H), 2.02–1.93 (m, 1H), 1.92–1.85 (m, 2H), 1.79–1.71 (m, 2H), 1.57–1.49 (m, 1H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.59, 159.19, 157.95, 154.76, 149.07, 147.80, 145.03, 144.83, 141.75, 141.44, 138.25, 134.77, 133.78, 132.26, 131.07, 130.06, 128.72, 127.54, 124.11, 123.23, 120.90, 120.33, 117.84, 116.43, 114.74, 114.69, 113.86, 69.82, 66.28, 63.22, 54.59, 53.00, 52.89, 50.47, 46.74, 38.07, 34.86, 29.72, 29.07, 28.43. ESI-MS m / z: 698.7 [M+H] + 。

[0240] Example 25

[0241] (R)-N-(4-(((4-(4-(3-(3-(3-Hydroxypyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-25)

[0242]

[0243] ESI-MS m / z: 712.5 [M+H] + 。

[0244] Example 26

[0245] N-(3-(((4-(4-(3-(3-(3,3-Difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-26)

[0246]

[0247] Step 1: 4-(4-(3-(3-(3,3-Difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine

[0248]

[0249] According to a preparation method similar to Steps 1 and 2 in Example 22, replacing (S)-3-hydroxypyrrolidine hydrochloride in Step 2 of Example 22 with 3,3-difluoropyrrolidine hydrochloride, this intermediate was prepared as a yellow solid with a yield of 30.2%. 11H NMR (600 MHz, Chloroform-d) δ 8.89 (d, J = 2.2 Hz, 1H), 8.77 (s, 1H), 8.44 (d, J = 8.2 Hz, 1H), 8.09 (d, J = 2.2 Hz, 1H), 7.37–7.34 (m, 2H), 7.12 (dd, J = 7.6, 1.0 Hz, 1H), 7.05–7.02 (m, 1H), 7.00–6.98 (m, 1H), 6.93–6.86 (m, 2H), 6.09 (d, J = 17.7 Hz, 1H), 5.61 (d, J = 11.0 Hz, 1H), 4.94 (t, J = 8.1 Hz, 2H), 4.08 (t, J = 6.2 Hz, 2H), 3.29 (t, J = 8.1 Hz, 2H), 2.95 (t, J = 13.1 Hz, 2H), 2.79 (t, J = 6.9 Hz, 2H), 2.69 (t, J = 7.2 Hz, 2H), 2.33–2.25 (m, 2H), 2.01 (p, J = 6.6 Hz, 2H). ESI-MS m / z: 514.5 [M+H] + 。

[0250] Step 2: 4-(4-(3-(3-(3,3-Difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0251]

[0252] According to a preparation method similar to that in Step 6 of Example 1, using 4-(4-(3-(3-(3,3-difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine to replace 4-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine to obtain this intermediate, which is a yellow solid with a yield of 58.6%. 11H NMR (400 MHz, Chloroform-d) δ 10.23 (s, 1H), 9.17 (d, J = 2.1 Hz, 1H), 8.79 (s, 1H), 8.46 (d, J = 2.1 Hz, 1H), 8.44 (dd, J = 8.3, 0.9 Hz, 1H), 7.33–7.26 (m, 2H), 7.08 (dd, J = 7.7, 1.0 Hz, 1H), 6.98–6.94 (m, 1H), 6.91 (dd, J = 2.6, 1.5 Hz, 1H), 6.88–6.82 (m, 1H), 4.93–4.86 (m, 2H), 4.01 (t, J = 6.2 Hz, 2H), 3.23 (t, J = 8.0 Hz, 2H), 2.89 (t, J = 13.2 Hz, 2H), 2.73 (s, 2H), 2.67–2.59 (m, 2H), 2.29–2.16 (m, 2H), 1.94 (p, J = 6.6 Hz, 2H). ESI-MS m / z: 516.2 [M+H] + 。

[0253] Step 4: N-(3-(((4-(4-(3-(3-(3,3-difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-26)

[0254]

[0255] Prepared by a similar preparation method to Example 2, using 4-(4-(3-(3-(3,3-difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the starting material. 11H NMR (600 MHz, DMSO-d6) δ 9.08 (s, 1H), 8.84 (d, J = 2.1 Hz, 1H), 8.69 (s, 1H), 8.60 (t, J = 5.5 Hz, 1H), 8.42 (d, J = 8.2 Hz, 1H), 8.13–8.00 (m, 2H), 7.67–7.55 (m, 3H), 7.40–7.29 (m, 2H), 7.13–6.99 (m, 3H), 6.96 (dd, J = 8.1, 2.6 Hz, 1H), 4.84 (t, J = 8.2 Hz, 2H), 4.07 (t, J = 6.3 Hz, 2H), 3.94 (s, 2H), 3.38 (q, J = 8.6, 7.4 Hz, 2H), 3.24 (t, J = 8.2 Hz, 2H), 2.89 (t, J = 13.4 Hz, 2H), 2.71–2.56 (m, 6H), 2.28–2.18 (m, 2H), 1.92–1.87 (m, 2H), 1.79–1.72 (m, 2H). 13 13C NMR (151 MHz, DMSO-d6) δ 163.53, 158.10, 156.93, 153.73, 148.05, 146.77, 143.97, 143.79, 140.72, 140.47, 137.20, 133.72, 132.73, 131.22, 130.04, 129.99 (t, J = 246.9 Hz), 129.03, 127.67, 126.51, 123.06, 122.16, 119.91, 119.27, 116.80, 115.39, 113.70, 113.64, 112.83, 65.03, 60.81 (t, J = 28.3 Hz), 53.55, 51.12 (t, J = 5.3 Hz), 51.09, 49.42, 45.69, 37.02, 34.68 (t, J = 23.8 Hz), 28.69, 28.02, 26.77. ESI-MS m / z: 718.3 [M+H] + 。

[0256] Example 27

[0257] N-(4-(((4-(4-(3-(3-(3,3-difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-27)

[0258]

[0259] Prepared by a similar preparation method as in Example 3, using 4-(4-(3-(3-(3,3-difluoropyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material to obtain this compound. ESI-MS m / z: 732.4 [M+H] + 。

[0260] Example 28

[0261] N-(3-(((4-(4-(2-methyl-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-28)

[0262]

[0263] Step 1: 3-(1H-indol-4-yl)-2-methylphenol

[0264]

[0265] 3-Bromo-2-methylphenol (5 g, 26.7 mmol), 4-indoleboronic acid pinacol ester (7.8 g, 32.1 mmol), Pd(dppf)Cl2 (0.39 g, 0.53 mmol), and potassium carbonate (9.24 g, 66.8 mmol) were successively added to a mixed solution of 1,4-dioxane (40 mL) and water (10 mL). The reaction was carried out at 90 °C for 3 h under N2 protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate multiple times. The filtrate was poured into 100 mL of water and extracted with ethyl acetate (100 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product as a light tan solid, which was directly used for the next reaction without purification. ESI-MS m / z: 224.2 [M+H] + 。

[0266] Step 2: 4-(4-(2-methyl-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0267]

[0268] ESI-MS m / z: 494.3 [M+H] + 。

[0269] According to a preparation method similar to steps 2 to 6 in Example 1, 3-(1H-indol-4-yl)-2-methylphenol was used to replace 3-(1H-indol-4-yl)phenol in step 2, thereby obtaining this key intermediate, which is a yellow solid. ESI-MS m / z: 224.2 [M+H] + 。

[0270] Step 3: N-(3-(((4-(4-(2-methyl-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-28)

[0271]

[0272] According to a preparation method similar to that in Example 2, this compound was prepared using 4-(4-(2-methyl-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material. ESI-MS m / z: 696.4 [M+H] + 。

[0273] Example 29

[0274] N-(4-(((4-(4-(2-methyl-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-29)

[0275]

[0276] According to a preparation method similar to that in Example 7, this compound was prepared using 4-(4-(2-methyl-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material. ESI-MS m / z: 710.4 [M+H] + 。

[0277] Example 30

[0278] N-(3-(((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-30)

[0279]

[0280] Step 1: 2-Chloro-3-(1H-indol-4-yl)phenol

[0281]

[0282] 3-Bromo-2-chlorophenol (10 g, 48.2 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (14.1 g, 57.8 mmol), Pd(dppf)Cl2 (0.71 g, 0.96 mmol), and potassium carbonate (16.7 g, 120.5 mmol) were successively added to a mixed solution of 1,4-dioxane (120 mL) and water (30 mL). The reaction was carried out at 90 °C for 3 h under N2 protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate several times. The filtrate was poured into 300 mL of water and extracted with ethyl acetate (300 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product as a light yellowish-brown solid, which was used directly in the next step without purification. 1 1H NMR (400 MHz, Chloroform-d) δ 8.17 (s, 1H), 7.39–7.34 (m, 1H), 7.22–7.18 (m, 2H), 7.17–7.13 (m, 1H), 7.06–7.02 (m, 1H), 7.02–6.96 (m, 2H), 6.29–6.25 (m, 1H). ESI-MS m / z: 244.1 [M+H] + 。

[0283] Step 2: 2-Chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol

[0284]

[0285] According to a similar preparation method as in Steps 2–4 of Example 1, in Step 2, 2-chloro-3-(1H-indol-4-yl)phenol was used instead of 3-(1H-indol-4-yl)phenol to obtain this key intermediate as a yellow solid. ESI-MS m / z: 401.2 [M+H] + 。

[0286] Step 3: 4-(4-(2-Chloro-3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0287]

[0288] According to a preparation method similar to Steps 5 and 6 in Example 1, in Step 5, 2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol was used to replace 3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol, thereby obtaining this key intermediate, which is a yellow solid. 1 HNMR(600MHz,Chloroform-d)δ10.23(s,1H),9.17(d,J=2.0Hz,1H),8.79(s,1H),8.54(d,J=8.2Hz,1H),8.46(d,J=2.1Hz,1H),7.34–7.28(m,1H),7.21(d,J=8.­0Hz,1H),6.97–6.90(m,2H),6.83(dd,J=7.5,1.4Hz,1H),4.96–4.85(m,2H),4.11(q,J=6.7Hz,2H),3.13–3.02(m,1H),2.97–2.90(m,1H),2.77–2.58(m,6H),2.13–2.08(m,2H),1.83–1.76(m,4H).ESI-MS m / z:514.4[M+H] + 。

[0289] Step 4: N-(3-(((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-30)

[0290]

[0291] According to a preparation method similar to that in Example 2, using 4-(4-(2-chloro-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material to obtain this compound. ESI-MS m / z:716.4[M+H] + 。

[0292] Example 31

[0293] N-(3-(((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)amino)propyl)imidazo[1,2-a]pyridine-7-carboxamide (Compound I-31)

[0294]

[0295] Prepared by a similar preparation method to Example 6, using 4-(4-(2-chloro-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material to obtain this compound. ESI-MS m / z: 716.4 [M+H] + 。

[0296] Example 32

[0297] N-(4-(((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-yl)methyl)amino)butyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-32)

[0298]

[0299] Prepared by a similar preparation method to Example 3, using 4-(4-(2-chloro-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material to obtain this compound. ESI-MS m / z: 730.4 [M+H] + 。

[0300] Example 33

[0301] N-((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-yl)methyl)-3-(1H-imidazol-1-yl)propan-1-amine (Compound I-33)

[0302]

[0303] Prepared by a similar preparation method to Example 9, using 4-(4-(2-chloro-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material to obtain this compound. 11H NMR (600 MHz, DMSO-d6) δ 8.86 (d, J = 2.1 Hz, 1H), 8.73 (s, 1H), 8.50 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 2.1 Hz, 1H), 7.59 (s, 1H), 7.40–7.31 (m, 2H), 7.19 (dd, J = 8.4, 1.4 Hz, 1H), 7.15 (d, J = 1.3 Hz, 1H), 6.98–6.91 (m, 2H), 6.85 (s, 1H), 4.91 (t, J = 8.7 Hz, 2H), 4.16 (q, J = 5.8 Hz, 2H), 4.04 (t, J = 7.0 Hz, 2H), 3.92 (s, 2H), 2.98 (t, J = 8.3 Hz, 2H), 2.70 (t, J = 7.3 Hz, 2H), 2.59 (s, 4H), 2.50–2.46 (m, 2H), 1.99 (p, J = 6.8 Hz, 2H), 1.88 (p, J = 6.9 Hz, 2H), 1.76–1.68 (m, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 156.98, 153.72, 153.70, 148.08, 146.75, 143.37, 140.27, 139.36, 136.63, 135.40, 132.79, 131.23, 130.95, 127.69, 127.11, 126.11, 123.44, 121.7, 119.73, 118.68, 117.24, 112.10, 66.33, 53.22, 52.97, 51.46, 49.30, 44.74, 43.32, 30.24, 27.30, 27.12, 22.45. ESI-MS m / z: 623.3 [M+H] + 。

[0304] Example 34

[0305] (E)-1-((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-34)

[0306]

[0307] Step 1: (4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine

[0308]

[0309] Using 4-(4-(2-chloro-3-(3-pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde obtained in Step 3 of Example 30 as the raw material, this intermediate was prepared by a similar preparation method as in Steps 1 to 4 of Example 18 and is a yellow solid. ESI-MS m / z: 515.4 [M+H] + 。

[0310] Step 2: (E)-1-((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-34)

[0311]

[0312] Using (4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as the starting material, this compound was prepared by a similar synthesis method as in Step 5 of Example 18 and is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ9.81(bs,1H),8.87(d,J=2.2Hz,1H),8.74(s,1H),8.58–8.49(m,2H),8.41(s,2H),8.08(d,J=2.1Hz,1H),7.41–7.32(m,2H),7.28–7.17(m,3H),6.99–6.92(m,2H),4.95–4.88(m,2H),4.74(s,2H),4.22–4.13(m,2H),2.98(t,J=8.3Hz,2H),2.85–2.62(m,6H),2.02(p,J=6.6Hz,2H),1.81–1.71(m,4H). 13 C NMR(151MHz,DMSO-d6)δ158.34,158.02,155.02,154.71,150.26,148.19,147.64,146.49,144.37,140.41,138.82,136.47,133.62,132.62,132.07,128.19,127.20,124.62,122.88,120.78,118.39,116.64,115.55,113.20,67.27,54.30,53.95,52.41,43.13,28.37,27.79,23.46.ESI-MSm / z:659.0[M+H]+ 。

[0313] Example 35

[0314] N-((4-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-35)

[0315]

[0316] Using (4-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as the starting material, this compound was prepared by a synthetic method similar to that of Example 19 and is a yellow solid. 1 HNMR(600MHz,DMSO-d6)δ9.42(t,J=5.8Hz,1H),9.24–9.21(m,1H),8.91(d,J=2.1Hz,1H),8.72(s,1H),8.51(d,J=8.2Hz,1H),8.11–8.07(m,2H),7.73(dd,J=9.5,1.8Hz,1H),7.67(d,J=1.3Hz,1H),7.65(d,J=9.4Hz,1H),7.38–7.31(m,2H),7.17(dd,J=8.4,1.4Hz,1H),6.96–6.92(m,2H),4.93–4.86(m,2H),4.76(d,J=5.7Hz,2H),4.17–4.12(m,2H),3.00–2.94(m,2H),2.61(t,J=7.1Hz,2H),2.50–2.44(m,4H),1.98–1.92(m,2H),1.71–1.66(m,4H). 13 CNMR(151MHz,DMSO-d6)δ165.15,158.00,154.94,154.80,148.31,147.71,145.10,144.37,140.39,139.79,136.49,134.93,133.50,132.49,132.04,129.22,128.14,127.18,124.59,123.21,122.76,120.78,119.76,118.36,116.64,114.84,113.14,67.48,54.27,54.07,52.59,40.94,28.49,28.35,23.56.ESI-MS m / z:659.0[M+H]+ 。

[0317] Example 36

[0318] (E)-N-((4-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-3-yl)acrylamide (Compound I-36)

[0319]

[0320] Using (4-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(pyridin-3-yl)acrylic acid as starting materials, this compound was prepared by a synthetic method similar to that of Example 19 and was a yellow solid. 1 HNMR(600MHz,Chloroform-d)δ8.80(t,J=3.2Hz,1H),8.75(dd,J=7.1,3.7Hz,2H),8.57(dd,J=10.1,6.3Hz,2H),8.01(d,J=7.5Hz,1H),7.79(d,J=8.2Hz,1H),7.69(dd,J=15.7,4.8Hz,1H),7.37–7.28(m,2H),7.25(s,1H),6.99–6.93(m,2H),6.89(d,J=7.6Hz,1H),6.64(d,J=7.7Hz,1H),6.57(dd,J=15.5,4.7Hz,1H),4.92(s,2H),4.84–4.74(m,2H),4.16(s,2H),3.15–2.92(m,2H),2.91–2.68(m,6H),2.24–2.15(m,2H),1.89(s,4H). 1313C NMR (151 MHz, DMSO-d6) δ 165.47, 158.00, 154.96, 154.60, 150.72, 149.70, 148.31, 147.71, 144.38, 140.42, 139.85, 136.58, 136.42, 134.50, 133.34, 132.43, 132.04, 131.05, 128.20, 127.20, 124.57, 124.44, 124.11, 123.00, 120.78, 118.38, 113.24, 67.03, 54.28, 53.78, 52.16, 40.48, 28.35, 26.98, 23.33. ESI-MS m / z: 645.9 [M+H] + 。

[0321] Example 37

[0322] (E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-37)

[0323]

[0324] Using (4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid as starting materials, this compound was prepared by a synthetic method similar to that of Example 19 and was a yellow solid. 1 1H NMR (600 MHz, DMSO-d6) δ 8.84 (d, J = 2.2 Hz, 1H), 8.73 (s, 1H), 8.70 (t, J = 6.0 Hz, 1H), 8.51 (d, J = 8.2 Hz, 1H), 8.09 (d, J = 2.4 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.63 (dd, J = 8.7, 2.4 Hz, 1H), 7.39–7.31 (m, 3H), 7.18 (dd, J = 8.4, 1.4 Hz, 1H), 6.98–6.91 (m, 2H), 6.50–6.42 (m, 4H), 4.90 (t, J = 9.1 Hz, 2H), 4.63 (d, J = 5.9 Hz, 2H), 4.15 (d, J = 6.8 Hz, 2H), 2.97 (t, J = 8.3 Hz, 2H), 2.71–2.52 (m, 6H), 1.97 (p, J = 6.7 Hz, 2H), 1.74–1.68 (m, 4H). 1313C NMR (151 MHz, DMSO-d6) δ 166.40, 161.07, 158.00, 154.92, 154.78, 150.18, 148.31, 147.72, 144.37, 140.39, 140.22, 137.79, 136.48, 135.09, 133.23, 132.38, 132.03, 128.15, 127.17, 124.56, 122.78, 120.78, 119.38, 118.34, 116.88, 113.15, 108.76, 67.45, 54.27, 54.03, 52.55, 41.80, 28.36, 28.34, 23.55. ESI-MS m / z: 661.0 [M+H] + 。

[0325] Example 38

[0326] 1 - ((4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-3-ylmethyl)urea (Compound I-38)

[0327]

[0328] (4-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine (100 mg, 0.19 mmol), phenyl(pyridin-3-ylmethyl)carbamate (66.5 mg, 0.29 mmol), and DIPEA (50.2 mg, 0.39 mmol) were successively added to 3 mL of DMF, and the reaction was carried out at 80 °C for 4 h. After the reaction was completed, the reaction solution was poured into 30 mL of water, and extracted with ethyl acetate (20 mL × 3). The organic phase was washed twice with water and once with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, the filtrate was evaporated to dryness, and the crude product was separated and purified by column chromatography to obtain 58 mg of a yellow solid with a yield of 46.0%. 1HNMR(600MHz, DMSO-d6) δ 8.80 (d, J = 2.2 Hz, 1H), 8.73 (s, 1H), 8.52–8.47 (m, 2H), 8.43 (dd, J = 4.8, 1.7 Hz, 1H), 7.98 (d, J = 2.1 Hz, 1H), 7.68–7.64 (m, 1H), 7.40–7.31 (m, 3H), 7.19 (dd, J = 8.4, 1.4 Hz, 1H), 6.98–6.92 (m, 2H), 6.83 (t, J = 6.1 Hz, 1H), 6.79 (t, J = 6.1 Hz, 1H), 4.92–4.88 (m, 2H), 4.47 (d, J = 6.0 Hz, 2H), 4.27 (d, J = 6.0 Hz, 2H), 4.15 (d, J = 6.8 Hz, 2H), 2.97 (t, J = 8.3 Hz, 2H), 2.71–2.53 (m, 6H), 1.98 (p, J = 6.6 Hz, 2H), 1.76–1.68 (m, 4H). 13 C NMR(151MHz, DMSO-d6) δ 158.59, 158.03, 154.88, 154.77, 149.08, 148.37, 148.26, 147.78, 144.41, 141.54, 140.42, 136.68, 136.48, 135.29, 132.87, 132.27, 132.03, 128.17, 127.18, 124.53, 123.84, 122.82, 120.79, 118.32, 113.17, 67.41, 54.28, 54.05, 52.54, 41.20, 41.14, 28.36, 28.24, 23.52. ESI-MS m / z: 649.0 [M+H] + 。

[0329] Example 39

[0330] (R,E)-1-((4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-39)

[0331]

[0332] Step 1: (R)-tert-butyl 3-((2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenoxy)methyl)pyrrolidine-1-carboxylate

[0333]

[0334] At room temperature, 2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol (2.0 g, 4.99 mmol), (R)-1-Boc-3-hydroxymethylpyrrolidine (1.1 g, 5.49 mmol), and triphenylphosphine (1.96 g, 7.48 mmol) were successively added to 20 mL of anhydrous tetrahydrofuran. Under N2 protection, DIAD (1.51 g, 7.48 mmol) was slowly added in an ice bath. After the addition was complete, the mixture was refluxed for 6 h. After the reaction was completed, the reaction solution was poured into 150 mL of water, and extracted with dichloromethane (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered by suction, and the filtrate was concentrated to obtain a crude yellow oil. 20 mL of ethyl acetate was added for pulping for 30 min, and a yellow solid was observed to precipitate. The solid was filtered by suction, and the filter cake was dried to obtain 2.4 g of a yellow solid with a yield of 82.4%. It was directly used in the next reaction without purification. 1 1H NMR (600 MHz, Chloroform-d) δ 8.88 (d, J = 2.1 Hz, 1H), 8.78 (s, 1H), 8.55 (d, J = 8.2 Hz, 1H), 8.09 (s, 1H), 7.71–7.64 (m, 2H), 7.56–7.53 (m, 1H), 7.49–7.44 (m, 2H), 7.38–7.34 (m, 1H), 6.99–6.85 (m, 4H), 6.08 (d, J = 17.7 Hz, 1H), 5.60 (d, J = 11.0 Hz, 1H), 5.03–4.89 (m, 2H), 4.04 (s, 2H), 3.71–3.58 (m, 1H), 3.54 (s, 1H), 3.44–3.33 (m, 1H), 3.33–3.25 (m, 1H), 3.19–3.06 (m, 1H), 3.05–2.92 (m, 1H), 2.83–2.72 (m, 1H), 2.19–2.08 (m, 1H), 1.96–1.81 (m, 1H), 1.47 (s, 9H). ESI-MS m / z: 584.2 [M+H] + 。

[0335] Step 2: (R)-4-(4-(2-chloro-3-(pyrrolidin-3-ylmethoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine

[0336]

[0337] (R)-tert-Butyl 3-((2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenoxy)methyl)pyrrolidine-1-carboxylate (2.3 g, 3.94 mmol) was dissolved in 20 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added. The reaction was carried out at 30 °C for 5 h. After the reaction was completed, the reaction solution was concentrated, the residue was added to water, and the pH was adjusted to 8 with an aqueous NaOH solution. A yellow solid was precipitated, filtered by suction, and the filter cake was dried to obtain 1.8 g of a yellow solid with a yield of 94.5%. 1 HNMR (600 MHz, DMSO-d6) δ 9.08 (d, J = 2.2 Hz, 1H), 8.73 (s, 1H), 8.50 (d, J = 8.2 Hz, 1H), 8.23 (d, J = 2.1 Hz, 1H), 7.43–7.38 (m, 1H), 7.38–7.33 (m, 1H), 7.21 (dd, J = 8.4, 1.4 Hz, 1H), 7.03–6.98 (m, 2H), 6.96–6.92 (m, 1H), 6.33 (d, J = 17.8 Hz, 1H), 5.65 (d, J = 11.1 Hz, 1H), 4.90 (t, J = 8.4 Hz, 2H), 4.16 (d, J = 37.9 Hz, 2H), 3.43–3.38 (m, 1H), 3.30–3.27 (m, 1H), 3.24–3.19 (m, 1H), 3.11–3.06 (m, 1H), 2.98 (t, J = 8.2 Hz, 2H), 2.83–2.77 (m, 1H), 2.17–2.10 (m, 1H), 1.87–1.80 (m, 1H). ESI-MS m / z: 484.3 [M+H] + 。

[0338] Step 3: (R)-4-(4-(2-Chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine

[0339]

[0340] At room temperature, (R)-4-(4-(2-chloro-3-(pyrrolidin-3-ylmethoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine (1.8 g, 3.72 mmol) was added to 20 mL of dichloromethane, and then aqueous formaldehyde solution (37%, 0.56 g, 18.6 mmol) and NaBH(OAc)3 (1.97 g, 9.3 mmol) were added successively. The reaction was carried out at room temperature for 30 min. After the reaction was completed, the reaction solution was evaporated to dryness, and the residue was separated and purified by column chromatography to obtain 1.7 g of a yellow solid with a yield of 91.8%. 11H NMR (600 MHz, Chloroform-d) δ 8.80 (d, J = 2.2 Hz, 1H), 8.71 (s, 1H), 8.46 (d, J = 8.2 Hz, 1H), 7.99 (d, J = 2.2 Hz, 1H), 7.30–7.26 (m, 1H), 7.20–7.18 (m, 1H), 6.91–6.87 (m, 2H), 6.85–6.83 (m, 1H), 6.82–6.77 (m, 1H), 6.01 (d, J = 17.6 Hz, 1H), 5.53 (d, J = 11.0 Hz, 1H), 4.92–4.83 (m, 2H), 3.99–3.93 (m, 2H), 3.09–3.01 (m, 1H), 2.94–2.82 (m, 2H), 2.81–2.74 (m, 1H), 2.73–2.58 (m, 4H), 2.40 (s, 3H), 2.15–2.08 (m, 1H). ESI-MS m / z: 498.2 [M+H] + 。

[0341] Step 4: (R)-4-(4-(2-Chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde

[0342]

[0343] Using (R)-4-(4-(2-Chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)-7-vinylpyrido[3,2-d]pyrimidine as the raw material, this intermediate was prepared by a preparation method similar to that in Step 6 of Example 1 and is a yellow solid. 1 1H NMR (600 MHz, Chloroform-d) δ 10.23 (s, 1H), 9.17 (d, J = 2.1 Hz, 1H), 8.79 (s, 1H), 8.54 (d, J = 8.2 Hz, 1H), 8.46 (d, J = 2.1 Hz, 1H), 7.33–7.28 (m, 1H), 7.22–7.20 (m, 1H), 6.94 (dd, J = 7.6, 1.0 Hz, 1H), 6.90 (dd, J = 8.3, 1.4 Hz, 1H), 6.84 (dd, J = 7.6, 1.4 Hz, 1H), 4.96–4.85 (m, 2H), 4.02–3.95 (m, 2H), 3.08 (d, J = 10.0 Hz, 1H), 2.96–2.89 (m, 2H), 2.84–2.65 (m, 4H), 2.45 (s, 3H), 2.19–2.12 (m, 1H), 1.79–1.70 (m, 1H). ESI-MS m / z: 500.3 [M+H]+ .

[0344] Step 5: (R)-(4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine

[0345]

[0346] Using (R)-4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbaldehyde as the raw material, this intermediate was prepared by a similar preparation method to Steps 1-4 in Example 18 and is a yellow solid. 1 HNMR(600MHz,Chloroform-d)δ8.71(d,J=2.2Hz,2H),8.46(d,J=8.2Hz,1H),7.98(d,J=2.1Hz,1H),7.30–7.26(m,1H),7.18(d,J=7.9Hz,1H),6.91–6.87(m,2H),6.83(dd,J=7.6,1.4Hz,1H),4.93–4.83(m,2H),4.05(s,2H),3.98–3.92(m,2H),3.10–3.01(m,1H),2.91(s,1H),2.82–2.72(m,2H),2.70–2.55(m,3H),2.37(s,3H),2.11–2.07(m,1H),1.69–1.64(m,1H).ESI-MS m / z:501.4[M+H] + .

[0347] Step 6: (R,E)-1-((4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-39)

[0348]

[0349] Using (R)-(4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as the raw material, this compound was prepared by a similar preparation method to Step 5 in Example 18 and is a yellow solid. 1HNMR(600MHz, DMSO-d6) δ 9.83(bs, 1H), 8.87(d, J = 2.2Hz, 1H), 8.74(s, 1H), 8.58–8.48(m, 2H), 8.40(s, 2H), 8.08(d, J = 2.2Hz, 1H), 7.40–7.3(m, 2H), 7.27–7.17(m, 3H), 6.99–6.92(m, 2H), 4.91(t, J = 8.2Hz, 2H), 4.74(d, J = 4.0Hz, 2H), 4.05(d, J = 22.8Hz, 2H), 2.98(t, J = 8.3Hz, 2H), 2.90–2.83(m, 1H), 2.80–2.60(m, 4H), 2.43(s, 3H), 2.09–2.00(m, 1H), 1.70–1.62(m, 1H). 13 C NMR(151MHz, DMSO-d6) δ 158.34, 158.01, 155.01, 154.71, 150.23, 148.19, 147.64, 146.54, 144.37, 140.42, 138.81, 136.43, 133.62, 132.61, 132.07, 128.18, 127.20, 124.63, 123.02, 120.82, 118.39, 116.64, 115.54, 113.37, 71.86, 58.70, 55.54, 54.30, 43.13, 41.70, 37.30, 28.37, 27.57. ESI-MS m / z: 645.0[M + H] + 。

[0350] Example 40

[0351] (R,E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-40)

[0352]

[0353] This compound was prepared from (R)-(4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid as starting materials according to a preparation method similar to that of Example 19, and it is a yellow solid. 1HNMR(600MHz, DMSO-d6) δ 8.84 (d, J = 2.1Hz, 1H), 8.74–8.69 (m, 2H), 8.51 (d, J = 8.2Hz, 1H), 8.09 (d, J = 2.4Hz, 1H), 8.01 (d, J = 2.1Hz, 1H), 7.63 (dd, J = 8.7, 2.4Hz, 1H), 7.40–7.31 (m, 3H), 7.20 (dd, J = 8.4, 1.4Hz, 1H), 6.99–6.92 (m, 2H), 6.50–6.42 (m, 4H), 4.90 (t, J = 8.6Hz, 2H), 4.63 (d, J = 5.9Hz, 2H), 4.10–3.99 (m, 2H), 2.97 (t, J = 8.3Hz, 2H), 2.91–2.83 (m, 1H), 2.79–2.63 (m, 4H), 2.43 (s, 3H), 2.08–2.00 (m, 1H), 1.70–1.62 (m, 1H). 13 C NMR(151MHz, DMSO-d6) δ 166.41, 161.06, 158.00, 154.93, 154.70, 150.18, 148.32, 147.72, 144.38, 140.42, 140.22, 137.81, 136.42, 135.11, 133.23, 132.38, 132.04, 128.17, 127.19, 124.57, 123.02, 120.82, 119.38, 118.36, 116.87, 113.35, 108.76, 71.84, 58.66, 55.51, 54.28, 41.66, 37.30, 28.35, 27.56. ESI-MS m / z: 647.0 [M+H] + 。

[0354] Example 41

[0355] (S,E)-1-((4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-41)

[0356]

[0357] Prepared by a similar method to Example 39, substituting (S)-1-Boc-3-hydroxymethylpyrrole for (R)-1-Boc-3-hydroxymethylpyrrole in Step 1 of Example 39 to obtain the compound. 1H NMR (600 MHz, DMSO-d6) δ 9.77 (bs, 1H), 8.87 (d, J = 2.2 Hz, 1H), 8.74 (s, 1H), 8.58–8.47 (m, 2H), 8.43–8.36 (m, 2H), 8.07 (d, J = 2.1 Hz, 1H), 7.39–7.31 (m, 2H), 7.27–7.17 (m, 3H), 6.98–6.92 (m, 2H), 4.90 (d, J = 7.5 Hz, 2H), 4.73 (s, 2H), 4.06–3.94 (m, 2H), 3.01–2.95 (m, 2H), 2.68–2.54 (m, 3H), 2.49–2.44 (m, 2H), 2.29 (s, 3H), 2.02–1.94 (m, 1H), 1.61–1.54 (m, 1H). 13 C NMR (151 MHz, DMSO-d6) δ 158.38, 158.01, 155.02, 154.80, 150.22, 148.19, 147.64, 146.63, 144.36, 140.40, 138.80, 136.47, 133.62, 132.62, 132.07, 128.16, 127.19, 124.63, 122.92, 120.82, 118.38, 116.68, 115.59, 113.34, 72.34, 59.21, 55.75, 54.30, 43.14, 42.12, 37.50, 28.37, 27.84. ESI-MS m / z: 644.9 [M+H] + 。

[0358] Example 42

[0359] (S,E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-42)

[0360]

[0361] Using (S)-(4-(4-(2-chloro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid as starting materials, this compound was prepared according to a preparation method similar to that of Example 19 and is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ8.84(d,J=2.2Hz,1H),8.74–8.70(m,2H),8.51(d,J=8.2Hz,1H),8.09(d,J=2.5Hz,1H),8.01(d,J=2.1Hz,1H),7.64(dd,J=8.7,2.5Hz,1H),7.40–7.31(m,3H),7.20(dd,J=8.4,1.4Hz,1H),6.99–6.92(m,2H),6.50–6.42(m,4H),4.90(t,J=8.7Hz,2H),4.63(d,J=5.9Hz,2H),4.10–3.98(m,2H),2.97(t,J=8.4Hz,2H),2.91–2.84(m,1H),2.80–2.61(m,4H),2.43(s,3H),2.10–2.00(m,1H),1.70–1.62(m,1H). 13 C NMR(151MHz,DMSO-d6)δ166.41,161.06,158.00,154.93,154.70,150.19,148.31,147.72,144.38,140.42,140.22,137.82,136.42,135.11,133.23,132.38,132.04,128.17,127.19,124.57,123.02,120.82,119.38,118.36,116.86,113.35,108.76,71.85,58.67,55.52,54.28,41.68,37.30,28.35,27.56.ESI-MS m / z:647.0[M+H] + 。

[0362] Example 43

[0363] (E)-2-cyano-1-((4-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-43)

[0364]

[0365] Step 1: 2-Fluoro-3-(1H-indol-4-yl)phenol

[0366]

[0367] Prepared according to a similar preparation method as in Step 1 of Example 30, substituting 3-bromo-2-fluorophenol for 3-bromo-2-chlorophenol to obtain this compound. ESI-MS m / z: 228.1 [M+H] + .

[0368] Step 2: 3-(1-(7-Bromopyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)-2-fluorophenol

[0369]

[0370] Prepared according to a similar preparation method as in Steps 2 and 3 of Example 1, substituting 3-bromo-2-fluorophenol for 3-(1H-indol-4-yl)phenol to obtain this compound. ESI-MS m / z: 437.1 [M+H] + .

[0371] Step 3: 4-(4-(2-Fluoro-3-hydroxyphenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbonitrile

[0372]

[0373] At room temperature, add 3-(1-(7-bromopyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)-2-fluorophenol (2.0 g, 4.57 mmol), zinc cyanide (0.54 g, 4.57 mmol), zinc powder (0.06 g, 0.91 mmol), Pd(dppf)Cl2 (0.13 g, 0.18 mmol), and Pd(dba)2·CHCl3 (0.19 g, 0.18 mmol) to 20 mL of N-methylpyrrolidone, protect with nitrogen, and react at 90 °C for 4 h. After the reaction is completed, filter with diatomaceous earth, wash the filter cake with a small amount of N-methylpyrrolidone, then pour the filtrate into 300 mL of water, filter, and dry to obtain 1.5 g of a yellow solid with a yield of 85.5%. ESI-MS m / z: 384.3 [M+H] + .

[0374] Step 4: (4-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)carbonitrile

[0375]

[0376] Using a preparation method similar to that of Step 5 in Example 1, 4-(4-(2-fluoro-3-hydroxyphenyl)indolin-1-yl)pyrido[3,2-d]pyrimidine-7-carbonitrile was used to replace 3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol to obtain this intermediate, which is a yellow solid. ESI-MS m / z: 495.3 [M+H] + 。

[0377] Step 5: (4-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine

[0378]

[0379] (4-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)carbonitrile (1.5 g, 3.01 mmol) and Raney nickel (0.11 g, 1.96 mmol) were added to 20 mL of ammonia-methanol solution, and the reaction was carried out at 40 °C for 2 h under a hydrogen atmosphere. After the reaction was completed, it was filtered by suction through diatomaceous earth, the filtrate was evaporated to dryness, and the crude product was separated and purified by column chromatography to obtain 0.85 g of a yellow solid with a yield of 56.3%. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 2H), 8.43 (d, J = 8.2 Hz, 1H), 7.98 (s, 1H), 7.28 (d, J = 15.6 Hz, 1H), 7.11–6.89 (m, 3H), 6.88–6.77 (m, 1H), 4.87 (t, J = 8.2 Hz, 2H), 4.16–3.97 (m, 4H), 3.08 (t, J = 7.9 Hz, 2H), 2.76–2.41 (m, 6H), 2.14–1.94 (m, 2H), 1.75 (s, 4H). ESI-MS m / z: 499.3 [M+H] + 。

[0380] Step 5: (E)-2-Cyano-1-((4-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-43)

[0381]

[0382] Using (4-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as a raw material, this compound was prepared by a synthesis method similar to that in Step 5 of Example 18 and is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ9.88(bs,1H),8.89(d,J=2.2Hz,1H),8.74(s,1H),8.60(s,1H),8.50(d,J=8.2Hz,1H),8.42(s,2H),8.09(d,J=2.2Hz,1H),7.39–7.34(m,1H),7.31–7.20(m,4H),7.05(d,J=7.6Hz,1H),7.03–6.97(m,1H),4.91(t,J=8.2Hz,2H),4.76(d,J=5.7Hz,2H),4.21(t,J=6.1Hz,2H),3.27–3.00(m,8H),2.23–2.14(m,2H),1.95–1.87(m,4H). 13 C NMR(151MHz,DMSO-d6)δ158.03,155.03,150.56,149.05(d,J=243.2Hz),148.24,147.65,147.11(d,J=10.8Hz),146.03,144.57,138.89,133.62,132.60,132.53,132.47,128.40(d,J=13.0Hz),127.37,125.05,124.91(d,J=4.3Hz),122.80,118.53,116.57,115.46,114.77,66.85,54.41,53.44,51.72,43.10,28.63,26.01,23.22.ESI-MS m / z:643.0[M+H] + 。

[0383] Example 44

[0384] N-((4-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-44)

[0385]

[0386] Using (4-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as the starting material, this compound was prepared according to a synthetic method similar to that of Example 19 and was a yellow solid. 1 HNMR(600MHz,DMSO-d6)δ9.43(t,J=5.7Hz,1H),9.22(d,J=1.6Hz,1H),8.92(d,J=2.1Hz,1H),8.73(s,1H),8.50(d,J=8.2Hz,1H),8.12–8.07(m,2H),7.72(dd,J=9.4,1.8Hz,1H),7.68–7.62(m,2H),7.39–7.33(m,1H),7.27–7.20(m,2H),7.04(d,J=7.5Hz,1H),7.02–6.97(m,1H),4.91(t,J=8.2Hz,2H),4.76(d,J=5.7Hz,2H),4.20(t,J=6.1Hz,2H),3.26–2.97(m,8H),2.19–2.11(m,2H),1.94–1.83(m,4H). 13 C NMR(151MHz,DMSO-d6)δ164.08,156.96,153.89,147.98(d,J=245.0Hz),147.35,146.65,146.07(d,J=10.8Hz),144.04,143.52,138.84,133.86,132.46,131.47,131.39,128.19,127.34(d,J=13.1Hz),126.30,123.96,123.84(d,J=4.5Hz),122.22,121.71,118.70,117.44,115.56,113.78,113.68,65.85,53.33,52.44,50.74,39.86,27.56,25.19,22.18.ESI-MS m / z:643.2[M+H] + 。

[0387] Example 45

[0388] (E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-45)

[0389]

[0390] Using (4-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid as starting materials, this compound was prepared by a synthetic method similar to that of Example 19 and is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ8.84(d,J=2.2Hz,1H),8.76–8.71(m,2H),8.49(d,J=8.2Hz,1H),8.09(d,J=2.4Hz,1H),8.02(d,J=2.1Hz,1H),7.64(dd,J=8.7,2.5Hz,1H),7.39–7.33(m,2H),7.26–7.19(m,2H),7.04(d,J=7.5Hz,1H),7.01–6.96(m,1H),6.50–6.42(m,4H),4.90(t,J=8.2Hz,2H),4.63(d,J=5.9Hz,2H),4.18(t,J=6.2Hz,2H),3.11–2.85(m,8H),2.13–2.06(m,2H),1.83(s,4H). 13 C NMR(151MHz,DMSO-d6)δ166.41,161.07,158.01,154.94,150.18,149.05(d,J=244.8Hz),148.36,147.73,147.21(d,J=10.8Hz),144.58,140.27,137.79,135.10,133.24,132.54,132.43,132.36,128.38(d,J=13.0Hz),127.35,125.00,124.88(d,J=4.4Hz),122.67,119.38,118.48,116.89,114.70,108.77,67.10,54.38,53.66,52.01,28.60,26.87,23.33.ESI-MSm / z:644.9[M+H] + 。

[0391] Example 46

[0392] (R,E)-2-Cyano-1-((4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-46)

[0393]

[0394] Step 1: 2-Fluoro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol

[0395]

[0396] According to a preparation method similar to that of intermediate 2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol, in Step 1 of Example 30, 3-bromo-2-fluorophenol was used to replace 3-bromo-2-chlorophenol, thereby obtaining this intermediate, which is a yellow solid. 1 H NMR(600MHz,DMSO-d6)δ9.97(s,1H),9.09(d,J=2.2Hz,1H),8.73(s,1H),8.47(d,J=8.1Hz,1H),8.23(d,J=2.2Hz,1H),7.37–7.33(m,1H),7.10–7.06(m,1H),7.04–6.97(m,3H),6.83–6.79(m,1H),6.33(d,J=17.8Hz,1H),5.65(d,J=11.2Hz,1H),4.90(t,J=8.2Hz,2H),3.09(t,J=8.2Hz,2H).ESI-MS m / z:385.2[M+H] + 。

[0397] Step 2: (R)-(4-(4-(2-Fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine

[0398]

[0399] According to a preparation method similar to Steps 1 to 5 in Example 39, in Step 1, 2-fluoro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol was used to replace 2-chloro-3-(1-(7-vinylpyrido[3,2-d]pyrimidin-4-yl)indolin-4-yl)phenol to obtain this intermediate, which is a yellow solid.ESI-MS m / z:485.4[M+H] + 。

[0400] Step 3: (R,E)-2-Cyano-1-((4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-46)

[0401]

[0402] Using (R)-(4-(4-(2-Fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine as a raw material, this compound was prepared according to a preparation method similar to that in Step 5 of Example 18 and is a yellow solid. 1 HNMR(600MHz,Chloroform-d)δ9.83(bs,1H),8.88(d,J=2.2Hz,1H),8.73(s,1H),8.56(s,1H),8.49(d,J=8.2Hz,1H),8.40(s,2H),8.08(d,J=2.1Hz,1H),7.38–7.33(m,1H),7.27–7.18(m,4H),7.04(d,J=7.6Hz,1H),7.00–6.95(m,1H),4.91(t,J=8.3Hz,2H),4.74(s,2H),4.08–3.99(m,2H),3.08(t,J=8.2Hz,2H),2.85(t,J=8.7Hz,1H),2.81–2.74(m,1H),2.73–2.62(m,3H),2.43(s,3H),2.07–1.99(m,1H),1.67–1.58(m,1H). 13 C NMR(151MHz,DMSO-d6)δ158.34,158.02,155.03,150.23,149.09(d,J=245.1Hz),148.23,147.65,147.32(d,J=10.9Hz),146.30,144.57,138.86,133.62,132.61,132.56,132.46,128.40(d,J=13.0Hz),127.35,125.07,124.88(d,J=4.4Hz),122.73,118.51,116.63,115.53,114.91,72.05,58.64,55.47,54.41,43.13,41.60,37.26,28.61,27.56.ESI-MS m / z:629.1[M+H] + 。

[0403] Example 47

[0404] (R,E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-47)

[0405]

[0406] This compound was prepared as a yellow solid from (R)-(4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methylamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid according to a synthetic method similar to that of Example 19. 1 H NMR(600MHz,DMSO-d6)δ8.86–8.82(m,1H),8.78–8.70(m,2H),8.49(d,J=8.2Hz,1H),8.09(d,J=2.4Hz,1H),8.02(s,1H),7.63(dd,J=8.7,2.4Hz,1H),7.39–7.33(m,2H),7.26–7.19(m,2H),7.06–6.97(m,2H),6.50–6.42(m,4H),4.93–4.86(m,2H),4.63(d,J=5.9Hz,2H),4.15–4.06(m,2H),3.23–3.16(m,1H),3.12–2.91(m,5H),2.86–2.78(m,1H),2.64(s,3H),2.18–2.09(m,1H),1.80–1.71(m,1H). 13 C NMR(151MHz,DMSO-d6)δ166.41,161.06,158.02,154.94,150.18,149.08(d,J=245.0Hz),148.36,147.73,147.17(d,J=11.1Hz),144.59,140.28,137.79,135.10,133.24,132.49,132.42,132.36,128.44(d,J=13.0Hz),127.35,125.01,124.89(d,J=4.5Hz),122.89,119.38,118.49,116.89,114.94,108.77,71.25,57.74,55.03,54.38,40.84,36.96,28.60,27.09.ESI-MS m / z:631.1[M+H] + 。

[0407] Example 48

[0408] (S,E)-2-Cyano-1-((4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-48)

[0409]

[0410] Prepared by a similar preparation method to Example 46, replacing (R)-1-Boc-3-hydroxymethylpyrrole with (S)-1-Boc-3-hydroxymethylpyrrole to obtain this compound. 1 HNMR(600MHz,DMSO-d6)δ9.84(bs,1H),8.88(d,J=2.1Hz,1H),8.74(s,1H),8.55(s,1H),8.50(d,J=8.2Hz,1H),8.41(s,2H),8.08(d,J=2.2Hz,1H),7.39–7.33(m,1H),7.27–7.17(m,4H),7.05(d,J=7.6Hz,1H),7.01–6.95(m,1H),4.91(t,J=8.2Hz,2H),4.74(s,2H),4.07–3.98(m,2H),3.08(t,J=8.2Hz,2H),2.80(t,J=8.7Hz,1H),2.75–2.57(m,4H),2.40(s,3H),2.06–1.99(m,1H),1.65–1.57(m,1H). 13 C NMR(151MHz,DMSO-d6)δ158.36,158.02,155.03,150.22,149.09(d,J=245.0Hz),148.22,147.65,147.35(d,J=11.0Hz),146.47,144.56,138.85,133.62,132.60,132.56,132.46,128.39(d,J=13.0Hz),127.35,125.07,124.87(d,J=4.4Hz),122.70,118.51,116.64,115.54,114.90,72.18,58.78,55.54,54.40,43.13,41.72,37.31,28.61,27.64.ESI-MS m / z:629.1[M+H] + 。

[0411] Example 49

[0412] (S,E)-3-(6-Aminopyridin-3-yl)-N-((4-(4-(2-fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methyl)acrylamide (Compound I-49)

[0413]

[0414] Using (S)-(4-(4-(2-Fluoro-3-((1-methylpyrrolidin-3-yl)methoxy)phenyl)indolin-1-yl)pyrido[3,2-d]pyrimidin-7-yl)methanamine and (E)-3-(6-aminopyridin-3-yl)acrylic acid as starting materials, this compound was prepared by a synthetic method similar to that of Example 19 and is a yellow solid. 1 HNMR(600MHz,DMSO-d6)δ8.84(d,J=2.2Hz,1H),8.74–8.69(m,2H),8.49(d,J=8.2Hz,1H),8.09(d,J=2.4Hz,1H),8.02(d,J=2.1Hz,1H),7.64(dd,J=8.7,2.4Hz,1H),7.39–7.33(m,2H),7.25–7.18(m,2H),7.04(d,J=7.5Hz,1H),7.00–6.95(m,1H),6.50–6.41(m,4H),4.90(t,J=8.2Hz,2H),4.63(d,J=5.9Hz,2H),4.06–3.97(m,2H),3.08(t,J=8.2Hz,2H),2.76(t,J=8.6Hz,1H),2.72–2.63(m,2H),2.61–2.52(m,2H),2.37(s,3H),2.05–1.97(m,1H),1.63–1.56(m,1H). 1313C NMR (151 MHz, DMSO-d6) δ 166.40, 161.07, 158.02, 154.95, 150.19, 149.08 (d, J = 244.9 Hz), 148.36, 147.74, 147.33 (d, J = 11.0 Hz), 144.59, 140.27, 137.82, 135.11, 133.24, 132.55, 132.43, 132.37, 128.40 (d, J = 13.2 Hz), 127.34, 125.02, 124.87 (d, J = 4.5 Hz), 122.72, 119.37, 118.47, 116.86, 114.89, 108.76, 72.11, 58.69, 55.49, 54.38, 41.65, 37.28, 28.60, 27.60. ESI-MS m / z: 631.2 [M+H] + 。

[0415] Example 50

[0416] N-(2-(((8-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-50)

[0417]

[0418] Step 1: 8-(4-(3-(3-(Diethylamino)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde

[0419]

[0420] According to a preparation method similar to Steps 1 to 6 in Example 1, in Step 3, 3-bromo-8-chloro-1,7-naphthyridine is used to replace 7-bromo-4-chloropyrido[3,2-d]pyrimidine, and in Step 6, 3-chloro-N,N-diethylpropan-1-amine is used to replace 1-(3-chloropropyl)pyrrolidine, thereby obtaining this intermediate, which is a red solid. 1HNMR(600MHz,DMSO-d6)δ10.27(s,1H),9.23–9.21(m,1H),8.92–8.86(m,1H),8.32–8.28(m,1H),7.58–7.55(m,2H),7.38–7.34(m,1H),7.20–7.16(m,1H),7.06(dd,J=7.3,2.0Hz,1H),7.02(d,J=2.0Hz,1H),6.96–6.92(m,2H),4.61–4.56(m,2H),4.06(t,J=6.3Hz,2H),3.24–3.18(m,2H),2.63–2.52(m,6H),1.91–1.81(m,2H),0.97(t,J=7.1Hz,6H).ESI-MS m / z:481.3[M+H] + 。

[0421] Step 2: N-(2-(((8-(4-(3-(3-(diethylamino)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-50)

[0422]

[0423] Prepared by a similar preparation method as in Step 9 of Example 1, using 8-(4-(3-(3-(diethylamino)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde as the raw material to obtain the target compound, which is a yellow solid. 11H NMR (600 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.88 (d, J = 2.1 Hz, 1H), 8.59–8.56 (m, 1H), 8.22 (d, J = 2.2 Hz, 1H), 8.17 (d, J = 5.4 Hz, 1H), 8.05 (s, 1H), 7.67–7.63 (m, 2H), 7.60 (d, J = 9.5 Hz, 1H), 7.50 (d, J = 8.1 Hz, 1H), 7.40–7.33 (m, 2H), 7.19–7.13 (m, 1H), 7.07 (d, J = 7.6 Hz, 1H), 7.05–7.02 (m, 1H), 6.97–6.90 (m, 2H), 4.55 (t, J = 8.2 Hz, 2H), 4.08 (t, J = 6.3 Hz, 2H), 3.98 (s, 2H), 3.44 (q, J = 6.3 Hz, 2H), 3.20 (t, J = 8.3 Hz, 2H), 2.77 (t, J = 6.5 Hz, 2H), 2.69–2.54 (m, 6H), 1.93–1.86 (m, 2H), 0.99 (t, J = 7.1 Hz, 6H). 13 13C NMR (151 MHz, DMSO-d6) δ 164.77, 159.14, 154.83, 149.99, 147.08, 145.06, 142.17, 141.88, 138.39, 137.99, 135.28, 134.82, 133.61, 133.25, 130.01, 129.94, 128.84, 127.18, 123.36, 121.50, 120.92, 120.32, 116.44, 114.69, 114.67, 114.30, 113.74, 66.11, 54.43, 50.42, 49.04, 48.42, 46.86, 29.50, 29.01, 26.50, 11.70. ESI-MS m / z: 669.4 [M+H] + 。

[0424] Example 51

[0425] N-(2-(((8-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-51)

[0426]

[0427] Step 1: 8-(4-(3-(3-(Pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde

[0428]

[0429] According to a preparation method similar to Steps 1 to 6 in Example 1, 3-bromo-8-chloro-1,7-naphthyridine is used to replace 7-bromo-4-chloropyrido[3,2-d]pyrimidine in Step 3, thereby obtaining this intermediate, which is a red solid. ESI-MS m / z: 479.4 [M+H] + 。

[0430] Step 2: N-(2-(((8-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)amino)ethyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-51)

[0431]

[0432] According to a preparation method similar to Step 9 in Example 1, the target compound is obtained using 8-(4-(3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)carbaldehyde as the raw material, which is a yellow solid. 1 H NMR (600 MHz, DMSO-d6) δ 9.11 (d, J = 1.7 Hz, 1H), 8.88 (d, J = 2.1 Hz, 1H), 8.57 (t, J = 5.7 Hz, 1H), 8.21 (d, J = 2.1 Hz, 1H), 8.17 (d, J = 5.4 Hz, 1H), 8.05 (s, 1H), 7.67–7.62 (m, 2H), 7.60 (d, J = 9.4 Hz, 1H), 7.50 (d, J = 8.1 Hz, 1H), 7.39–7.33 (m, 2H), 7.19–7.13 (m, 1H), 7.09–7.05 (m, 1H), 7.05–7.02 (m, 1H), 6.97–6.90 (m, 2H), 4.55 (t, J = 8.3 Hz, 2H), 4.09 (t, J = 6.4 Hz, 2H), 3.98 (s, 2H), 3.44 (q, J = 6.3 Hz, 2H), 3.20 (t, J = 8.3 Hz, 2H), 2.76 (t, J = 6.5 Hz, 2H), 2.66–2.61 (m, 2H), 2.57–2.52 (m, 4H), 1.96–1.90 (m, 2H), 1.72–1.69 (m, 4H). 1313C NMR (151 MHz, DMSO-d6) δ 164.77, 159.15, 154.83, 149.98, 147.07, 145.06, 142.17, 141.86, 138.45, 137.99, 135.27, 134.80, 133.61, 133.21, 130.00, 129.94, 128.83, 127.17, 123.38, 121.50, 120.90, 120.33, 116.43, 114.69, 114.65, 114.30, 114.28, 113.73, 66.24, 54.43, 54.04, 52.65, 50.44, 48.43, 29.00, 28.46, 23.53, 11.72. ESI-MS m / z: 667.3 [M+H] + 。

[0433] Example 52

[0434] (E)-1-((8-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-52)

[0435]

[0436] Step 1: 8-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde

[0437]

[0438] Following a similar preparation method to Steps 1 to 3 in Example 30, 3-bromo-8-chloro-1,7-naphthyridine was used to replace 7-bromo-4-chloropyrido[3,2-d]pyrimidine in Step 2, thereby obtaining this intermediate as an orange-red solid. 11H NMR (400 MHz, Chloroform-d) δ 10.21 (s, 1H), 9.19 (d, J = 2.1 Hz, 1H), 8.43 (d, J = 2.2 Hz, 1H), 8.26 (d, J = 5.5 Hz, 1H), 7.71 (d, J = 8.1 Hz, 1H), 7.21 (d, J = 5.5 Hz, 1H), 7.17 (d, J = 7.8 Hz, 2H), 6.89 (dd, J = 8.3, 1.4 Hz, 1H), 6.83 (dd, J = 7.7, 1.4 Hz, 1H), 6.78 (d, J = 7.5 Hz, 1H), 4.72–4.56 (m, 2H), 4.10 (d, J = 7.0 Hz, 2H), 3.05–2.87 (m, 2H), 2.72 (t, J = 7.5 Hz, 2H), 2.60 (d, J = 5.9 Hz, 4H), 2.07 (q, J = 6.9 Hz, 2H), 1.79–1.75 (m, 4H). ESI-MS m / z: 513.2 [M+H] + 。

[0439] Step 2: 8-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine

[0440]

[0441] Using 8-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde as the raw material, this intermediate was prepared by a method similar to Steps 1 to 4 in Example 18 and was a yellow oil. 1 1H NMR (400 MHz, Chloroform-d) δ 8.75 (d, J = 2.2 Hz, 1H), 8.15 (d, J = 5.5 Hz, 1H), 7.95–7.91 (m, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.17–7.08 (m, 3H), 6.91–6.80 (m, 2H), 6.75–6.69 (m, 1H), 4.56 (d, J = 31.6 Hz, 2H), 4.07 (d, J = 17.6 Hz, 4H), 3.09–2.86 (m, 2H), 2.70 (t, J = 7.5 Hz, 2H), 2.57 (s, 4H), 2.06 (q, J = 6.9 Hz, 2H), 1.81–1.71 (m, 4H). ESI-MS m / z: 514.3 [M+H] + 。

[0442] Step 3: (E)-1-((8-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)-2-cyano-3-(pyridin-4-yl)guanidine (Compound I-52)

[0443]

[0444] Using 8-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine as the starting material, this compound was prepared by a synthetic method similar to that in Step 5 of Example 18 and is a yellow solid. 1 HNMR(400MHz,DMSO-d6)δ9.89(bs,1H),8.88(d,J=2.2Hz,1H),8.55(s,1H),8.47–8.34(m,2H),8.21(d,J=5.5Hz,2H),7.63(d,J=8.1Hz,1H),7.47–7.32(m,2H),7.31–7.14(m,4H),6.99(d,J=7.6Hz,1H),6.76(dd,J=11.6,6.9Hz,1H),4.74(d,J=5.4Hz,2H),4.62(t,J=8.4Hz,2H),4.21(t,J=6.1Hz,2H),3.18–3.05(m,6H),2.92(t,J=8.5Hz,2H),2.16(p,J=6.4Hz,2H),1.94–1.83(m,4H). 13 C NMR(101MHz,DMSO-d6)δ158.24,154.75,154.45,150.15,148.95,146.41,145.03,142.05,140.85,136.12,135.85,135.25,133.47,132.94,130.88,128.07,126.74,123.12,121.96,120.78,116.60,115.44,114.76,114.27,113.06,66.81,54.13,53.54,51.90,43.37,28.25,26.31,23.21.ESI-MS m / z:658.9[M+H] + 。

[0445] Example 53

[0446] N-((8-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-53)

[0447]

[0448] Using 8-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine as the starting material, this compound was prepared according to a synthetic method similar to that of Example 19 and is a yellow solid. ESI-MS m / z: 658.0 [M+H] + 。

[0449] Example 54

[0450] (E)-3-(6-Aminopyridin-3-yl)-N-((8-(4-(2-chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)acrylamide (Compound I-54)

[0451]

[0452] Using 8-(4-(2-Chloro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine and (E)-3-(pyridin-3-yl)acrylic acid as the starting materials, this compound was prepared according to a synthetic method similar to that of Example 19 and is a yellow solid. ESI-MS m / z: 660.7 [M+H] + 。

[0453] Example 55

[0454] (E)-2-Cyano-1-((8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-55)

[0455]

[0456] Step 1: 8-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde

[0457]

[0458] According to a preparation method similar to Steps 1 to 3 in Example 30, 3-fluoro-2-chlorophenol was used to replace 3-bromo-2-chlorophenol in Step 1, and 3-bromo-8-chloro-1,7-naphthyridine was used to replace 7-bromo-4-chloropyrido[3,2-d]pyrimidine in Step 2, thereby obtaining this intermediate, which is an orange-red solid. 1 H NMR(400MHz,Chloroform-d)δ10.21(s,1H),9.20(d,J=2.1Hz,1H),8.43(d,J=2.2Hz,1H),8.26(d,J=5.5Hz,1H),7.64(d,J=8.1Hz,1H),7.22(d,J=5.5Hz,1H),7.18–7.13(m,1H),7.06–7.00(m,1H),6.96–6.89(m,1H),6.89–6.82(m,2H),4.62(t,J=8.2Hz,2H),4.09(t,J=6.3Hz,2H),3.07(t,J=8.2Hz,2H),2.77–2.69(m,2H),2.63(s,4H),2.08(p,J=6.6Hz,2H),1.85–1.74(m,4H).ESI-MS m / z:497.3[M+H] + 。

[0459] Step 2: 8-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine

[0460]

[0461] Using 8-(4-(2-Fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridine-3-carbaldehyde as the raw material, this intermediate was prepared according to a preparation method similar to Steps 1 to 4 in Example 18, and it is a yellow oil. 11H NMR (400 MHz, Chloroform-d) δ 8.76 (d, J = 2.2 Hz, 1H), 8.15 (d, J = 5.5 Hz, 1H), 7.96–7.93 (m, 1H), 7.42 (d, J = 8.1 Hz, 1H), 7.15–7.08 (m, 2H), 7.06–7.01 (m, 1H), 6.94–6.84 (m, 2H), 6.80 (d, J = 7.6 Hz, 1H), 4.54 (t, J = 8.3 Hz, 2H), 4.09 (t, J = 6.1 Hz, 2H), 4.05 (s, 2H), 3.05 (t, J = 8.2 Hz, 2H), 2.94–2.77 (m, 6H), 2.21–2.14 (m, 2H), 1.93–1.84 (m, 4H). ESI-MS m / z: 498.4 [M+H] + 。

[0462] Step 3: (E)-2-Cyano-1-((8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)-3-(pyridin-4-yl)guanidine (Compound I-55)

[0463]

[0464] Using 8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methylamine as the starting material, this compound was prepared by a synthetic method similar to that in Step 5 of Example 18 and is a yellow solid. ESI-MS m / z: 642.5 [M+H] + 。

[0465] Example 56

[0466] N-((8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)imidazo[1,2-a]pyridine-6-carboxamide (Compound I-56)

[0467]

[0468] Using 8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methylamine as the starting material, this compound was prepared by a synthetic method similar to that in Example 19 and is a yellow solid. ESI-MS m / z: 642.0 [M+H] + 。

[0469] Example 57

[0470] (E)-3-(6-Aminopyridin-3-yl)-N-((8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methyl)acrylamide (Compound I-57)

[0471]

[0472] This compound was prepared from 8-(4-(2-fluoro-3-(3-(pyrrolidin-1-yl)propoxy)phenyl)indolin-1-yl)-1,7-naphthyridin-3-yl)methanamine and (E)-3-(pyridin-3-yl)acrylic acid as starting materials according to a synthetic method similar to that of Example 19, and it is a yellow solid. ESI-MS m / z: 644.5 [M+H] + 。

[0473] Pharmacological Activity Study of the Compounds of the Present Invention

[0474] 1. Evaluation of the inhibitory activity of the compounds of the examples against the PD-1 / PD-L1 interaction

[0475] The homogeneous time-resolved fluorescence (HTRF) assay was used to detect the ability of the compounds of the present invention to inhibit the PD-1 / PD-L1 interaction. The detection kit was purchased from CisBio (CAT#64PD1PEG), which contains experimental reagents such as Anti-Tag1 Eu Cyptate, Anti-Tag2 XL665, Tag1-PD-L1, Tag2-PD-1, and Detection Buffer.

[0476] Experimental procedure:

[0477] (1) Dilute the compound to be tested with DMSO to 1 mM to obtain a stock solution, and continue to dilute the compound stock solution with DMSO in a 4-fold dilution series to obtain 6-8 concentrations. Dilute each concentration of the compound solution 20-fold with Detection buffer to obtain the working solution of the compound at the test concentration.

[0478] (2) Dilute the Tag2-PD-1 and Tag1-PD-L1 stock solutions 40-fold with Detection Buffer.

[0479] (3) Add 2 μL of the compound working solution, 4 μL of Tag2-PD-1, and 4 μL of Tag1-PD-L1 solution to a 96-well plate in sequence, mix well, and incubate at room temperature for 15 min.

[0480] (4) Dilute the Anti-Tag1 Eu Cryptate and Anti-Tag2 XL665 stock solutions 50-fold with Detection buffer, mix them in equal volumes, add 10 μL of this mixture to each reaction well, seal the membrane, and incubate at room temperature for 1 h.

[0481] (5) Detect the fluorescence signal using a SpectraMax i3X multimode microplate reader (excitation at 320 nm, emission at 665 nm and 620 nm).

[0482] (6) Data processing

[0483] Emission Ratio (ER) = 665 nm Emission signal / 620 nm Emission signal

[0484] Inhibition rate = (ER positive - ER sample ) / (ER positive - ER negative ) × 100%

[0485] Wherein, ER positive : Fluorescence emission ratio of the positive control; ER sample : Fluorescence emission ratio of the test compound; ER negative : Fluorescence emission ratio of the negative control.

[0486] Finally, input the data using GraphPad Prism 8.0 software and plot the inhibition rate-concentration curve to calculate IC 50 .

[0487] The activity results of the compounds inhibiting the PD-1 / PD-L1 interaction are shown in Table 1, and the data represent the activity range of the compounds inhibiting the PD-1 / PD-L1 interaction or the IC 50 value. The ranges are as follows: A = 1 nM - 100 nM; B = 100.01 nM - 1 μM.

[0488] 2. Evaluation of the inhibitory activity of the example compounds against NAMPT

[0489] Experimental procedure:

[0490] (1) Prepare a 50 μL mixed system for the enzymatic reaction: 50 mM Tris-HCl (pH = 8.0), 12.5 mM MgCl2, 20 μM NAM, 0.4 mM phosphoribosyl pyrophosphate, 2 mM ATP, 30 μg / mL alcohol dehydrogenase, 10 μg / mL NMNAT, 1.5% alcohol, 1 mM DTT, 0.02% BSA, 0.01% Tween 20, and the test compound. Note that the DMSO content in the system should be less than 1%.

[0491] (2) Incubate the enzymatic reaction at 30 °C for 90 min, and test 8 - 12 concentrations for each compound.

[0492] (3) Detect the fluorescence signal with a Tecan Infinite M1000 microplate reader (excitation at 360 nm, emission at 460 nm).

[0493] (4) Data processing

[0494] Inhibition rate = (F - F negative ) / (F positive - F negative ) × 100%

[0495] Wherein, F positive : Fluorescence intensity without adding the drug; F negative : Fluorescence intensity without adding NAMPT;

[0496] Finally, input the data into GraphPad Prism 8.0 software and plot the inhibition rate-concentration curve to calculate IC 50 .

[0497] The activity results of the compound against NAMPT are shown in Table 1, and the data represent the activity range or IC 50 value of the compound inhibiting NAMPT. The ranges are as follows: A = 1 nM - 100 nM; B = 100.01 nM - 1 μM.

[0498] 3. Evaluation of the anti-proliferative activity of the example compound against human ovarian cancer cell line A2780

[0499] Experimental procedure: Seed the cells in the logarithmic growth phase at 2.1×10 4Inoculate into a 96-well plate at [number of cells] / mL, with the volume of the cell suspension being 100 μL / well, and culture under standard conditions. After 24 h, add 100 μL of different concentrations of the drug solution prepared with the culture medium to each well, set four replicates for each concentration, and incubate for 72 h under standard conditions. After the incubation is completed, take out the 96-well plate, add 50 μL of MTT solution (2 mg / mL) to each well, continue to incubate for 4 h and then take out. After discarding the supernatant by flicking the plate, add 100 μL of DMSO solution to each well, and place it on a micro oscillator and shake for 5 min to completely dissolve the crystals. Measure the absorbance (OD) of each well at 570 nm on an enzyme-linked immunosorbent assay (ELISA) reader.

[0500] Calculate the inhibition rate (IR) of cell proliferation in vitro according to the formula:

[0501] IR% = (OD control - OD sample ) / (OD control - OD blank ) × 100%

[0502] In the formula, OD control : OD value of the well without adding the drug; OD sample : OD value of the well with the added drug; OD blank : OD value of the well with only DMSO added.

[0503] Finally, input the data into GraphPad Prism 8.0 software and draw the inhibition rate-concentration curve to calculate IC 50 .

[0504] The results of the inhibition of A2780 cell proliferation activity by the compounds in the examples are shown in Table 1, and the data represent the activity range of the compounds inhibiting A2780 cell proliferation or the IC 50 value. The ranges are as follows: A = 1 nM - 100 nM; B = 100.01 nM - 1 μM; C = 1 μM - 10 μM.

[0505] The experimental results show that the compounds in the examples have significant inhibitory activity against the PD-1 / PD-L1 protein / protein interaction and NAMPT, and have significant anti-proliferative activity against A2780 cells.

[0506] Table 1 Results of the inhibition of PD-1 / PD-L1 interaction, NAMPT and A2780 cell proliferation activity by the compounds in the examples

[0507]

[0508]

Claims

1. A compound containing an indoline structure, its stereoisomers, and pharmaceutically acceptable salts, characterized in that: The structural formula is as shown in General Formula I: Wherein, X is selected from CH or N; R1 is selected from hydrogen, methyl or halogen; R2 is selected from R4 and R5 are each independently selected from hydrogen, (C1–C4) alkyl or hydroxy(C1–C4) alkyl, or R4, R5 and the nitrogen atom to which they are attached together form a 4- to 6-membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may optionally be substituted by 1 to 3 R7; R6 is independently selected from hydrogen, (C1–C4) alkyl, (C1–C4) alkylcarbonyl or (C1–C4) alkylsulfonyl; R7 is independently selected from hydrogen, halogen, hydroxy, carboxy, (C1–C4) alkyl or hydroxy(C1–C4) alkyl; m is an integer selected from 1 to 3; n is an integer selected from 0 to 5, wherein, When n is 0, R3 is selected from: When n is 1 to 5, R3 is selected from:

2. The indoline structure-containing compound according to claim 1, its stereoisomers and pharmaceutically acceptable salts, characterized in that: X is selected from CH or N; R1 is selected from hydrogen, methyl, fluorine or chlorine; R2 is selected from R4 and R5 are each independently selected from hydrogen or (C1–C4) alkyl, or R4, R5 and the nitrogen atom to which they are attached together form a 4- to 6-membered nitrogen-containing heterocycle; the nitrogen-containing heterocycle may optionally be substituted by 1 to 3 R7; R6 is independently selected from hydrogen or (C1–C4) alkyl; R7 is independently selected from hydrogen, halogen, hydroxy, carboxy or hydroxy(C1–C4) alkyl; m is an integer selected from 1 to 3; n is an integer selected from 0 to 5, wherein, When n is 0, R3 is selected from: When n is 1 to 5, R3 is selected from:

3. The indoline structure-containing compound according to any one of claims 1 or 2, its stereoisomers and pharmaceutically acceptable salts, characterized in that: Selected from: Selected from:

4. The compound containing indoline structure, its stereoisomers and pharmaceutically acceptable salts according to claim 1, characterized in that: The indoline structure-containing compound is one of the compounds having the following chemical structural formulas I-1 to I-57:

5. The preparation method of the indoline structure-containing compound according to any one of claims 1 to 4, characterized in that: The preparation method of the indoline structure-containing compound when n = 1 to 5 includes the following steps: (a) Using a 4-arylindole as a raw material, and obtaining Intermediate 2 under the action of a reducing agent such as sodium cyanoborohydride; (b) Using Intermediate 2 as a raw material, and carrying out an aromatic nucleophilic substitution reaction with 7-bromo-4-chloropyrido[3,2-d]pyrimidine or 3-bromo-8-chloro-1,7-naphthyridine under acid catalysis to obtain Intermediate 3; (c) Using Intermediate 3 as a raw material, and carrying out a coupling reaction with vinylboronic acid pinacol ester or tributylvinyltin reagent to obtain Intermediate 4; (d) Using Intermediate 4 as a raw material, and carrying out a nucleophilic substitution reaction with a small molecule halide or a Mitsunobu reaction with an alcohol compound to obtain Intermediate 5; (e) Using Intermediate 5 as a raw material, and obtaining Intermediate 6 under the action of an osmium reagent and an oxidizing agent; (f) Using Intermediate 6 as a raw material, and carrying out a reductive amination reaction with an amine compound under the action of sodium cyanoborohydride or sodium triacetoxyborohydride to obtain the target compound in General Formula I (n = 1 to 5); Alternatively, the preparation method of the indoline structure-containing compound when n = 1 to 5 includes the following steps: (g) Using Intermediate 6 as a raw material, and reacting with a mono-Boc-alkyl diamine compound under the action of sodium cyanoborohydride or sodium triacetoxyborohydride to obtain Intermediate 7; (h) Using intermediate 7 as a raw material, a deprotecting agent is used under acidic conditions to obtain intermediate 8; (i) Using intermediate 8 as a raw material, the target compound in general formula I (n = 1 - 5) is obtained through a nucleophilic substitution or amidation reaction; The preparation method of the indoline structure compound when n = 0 includes the following steps: (j) Using intermediate 6 as a raw material, intermediate 9 is obtained under the action of a reducing agent such as sodium borohydride; (k) Using intermediate 9 as a raw material, intermediate 10 is obtained under the action of a chlorinating agent such as thionyl chloride; (l,m) Using intermediate 10 as a raw material, intermediate 12 is obtained through a Gabriel reaction; (n) Using intermediate 12 as a raw material, the target compound in general formula I (n = 0) is obtained through a nucleophilic substitution or amidation reaction; Alternatively, the preparation method of the indoline structure compound when n = 0 includes the following steps: (o) Using intermediate 3 as a raw material, intermediate 13 is obtained through a coupling reaction with a cyanide reagent such as copper(I) cyanide or zinc cyanide; (p) Using intermediate 13 as a raw material, intermediate 14 is obtained through a nucleophilic substitution reaction with a small molecule halide or a Mitsunobu reaction with an alcohol compound; (q) Using intermediate 14 as a raw material, intermediate 12 is obtained through borane reduction or catalytic hydrogenation reaction; (r) Using intermediate 12 as a raw material, the target compound in general formula I (n = 0) is obtained through a nucleophilic substitution or amidation reaction.

6. The compound according to any one of claims 1 to 4, its stereoisomers and pharmaceutically acceptable salts, characterized in that: The pharmaceutically acceptable salts include salts formed with inorganic acids, organic acids or alkali metal ions, wherein, The inorganic acids are selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid or phosphoric acid; the organic acids are selected from: succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid or p-toluenesulfonic acid; the alkali metal ions are selected from lithium ions, sodium ions or potassium ions.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the compound according to any one of claims 1 - 4, its stereoisomers and pharmaceutically acceptable salts, carriers or excipients.

8. Use of the compound according to any one of claims 1 - 4, its stereoisomers and pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for preventing and / or treating diseases related to PD-1 / PD-L1 protein / protein interaction and NAMPT.

9. The application according to claim 8, wherein: The diseases related to PD-1 / PD-L1 protein / protein interaction and NAMPT are selected from cancer or infectious diseases.

10. The application according to claim 9, characterized in that: The cancer is selected from lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell carcinoma, glioma, neuroblastoma, squamous cell carcinoma of the lung, adenocarcinoma of the lung, bladder cancer, gastric cancer, colon cancer, colorectal cancer, kidney cancer, cholangiocarcinoma, gastric cancer, esophageal squamous cell carcinoma, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, skin cancer, epithelial cell carcinoma, leukemia or cervical cancer; the infectious diseases are selected from bacterial infections or viral infections.

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