3-Cyano-substituted quinoline compounds, their pharmaceutical compositions and uses

Abstract

The present invention discloses a 3-cyano-substituted quinoline compound, its pharmaceutical composition and application. The structure of this kind of compound is as shown in Formula I, and it also includes its pharmaceutically acceptable salts. It has excellent HPK1 inhibitory activity, good water solubility and pharmacokinetic stability, has the potential to be developed into drugs, and can be used in the preparation of drugs for preventing and / or treating cancer, inflammatory diseases, autoimmune diseases or immune-mediated diseases.

Description

Technical Field

[0001] The present invention relates to a 3-cyano-substituted quinoline compound, its pharmaceutical composition and application, and particularly to a 3-cyano-substituted quinoline compound with anti-tumor activity, its pharmaceutical composition and application. Background Art

[0002] Highly effective and low-toxic anti-tumor targeted drugs against specific targets have achieved great success in tumor treatment and become a hot spot in the research and development of new anti-cancer drugs. However, tumors can develop multi-drug resistance to targeted therapy drugs, rendering the drugs ineffective. In recent years, tumor immunotherapy (IO) as an adjuvant therapy can be used to reverse multi-drug resistance caused by immune system failure and has received extensive attention in the medical field. Tumor immunotherapy mainly refers to using the human immune mechanism to actively or passively enhance the immune function of patients, thereby killing tumor cells. Currently, there are two cancer immunotherapy strategies for reversing multi-drug resistance caused by immune system failure. One is to regulate the antigen signal related to T cell co-receptors, and the other is to enhance the recognition of tumor cells.

[0003] Hematopoietic progenitor kinase 1 (HPK1), also known as mitogen-activated protein kinase 1 (MAP4K1), is a serine / threonine protein kinase that uses ATP or other nucleotides as phosphate donors and belongs to the Ste20-related protein kinase MAP4K family. The MAP4K family also includes glucokinase (GCK / MAP4K2), GPK-like kinase (GLK / MAP4K3), GPK-like kinase (HGK / MAP4K4), SPS1 / STE20 homologous kinase (KHS / MAP4K5), and Nck-related kinase (MINK / MAP4K6). The kinases in the MAP4K family have highly similar protein structures and play important roles in regulating cell survival, migration, apoptosis, and autophagy. And HPK1 in the MAP4K family kinases, as a negative regulator of cellular immunity, participates in various cellular immune responses mediated by T cells, B cells, and dendritic cells and plays an important role in the body's immune response. HPK1 is a negative signal regulatory kinase of the T cell receptor (TCR). When the TCR is activated, cytoplasmic HPK1 is recruited to the vicinity of the cell membrane. Activated HPK1 phosphorylates the adaptor protein SLP76. Activated SLP76 serves as a docking site for the negative regulatory protein 14-3-3π, ultimately leading to the instability of the TCR signal complex, thereby downregulating the TCR signal.

[0004] Current research has found that HPK1 is mainly expressed in hematopoietic tissues and cells, and its expression in other tissues varies greatly. HPK1 participates in many signal cascades, including growth factor signals, MAPK signals, cytokine signals, apoptosis signals, growth factor signals, and antigen receptor signals. For example, after transfection into COS1 cells, HPK1 can specifically activate the SAPK / JNK pathway. By inhibiting the function of HPK1 or knocking out the HPK1 gene, the killing ability of immune cells against tumor cells can be enhanced, and HPK1 inhibitors have great potential in improving T cell function, antigen presentation, and counteracting the immunosuppressive tumor microenvironment. HPK1 inhibitors can achieve immune activation by blocking the kinase action of HPK1, thereby enhancing the efficacy of PD-1 / PD-L1 targeted therapeutic drugs.

[0005] In summary, targeted anti-tumor drugs represented by protein kinase inhibitors have become the mainstream of anti-tumor drug research and development at home and abroad; due to its important role in anti-tumor immune response, HPK1 has become a new target for tumor immunotherapy. At present, the research on HPK1 inhibitors is still in its infancy. Developing highly efficient and highly selective small molecule kinase inhibitors of HPK1 for clinical monotherapy or in combination with other immunotherapies, such as PD-1 monoclonal antibody, to improve the response rate and efficiency of tumor immunotherapy has very important scientific research significance and social value; however, there is currently no HPK1 inhibitor drug on the market, and there is an urgent need to develop new drug candidate compounds with novel structures and better pharmacodynamic effects to provide support for further new drug research and development. Summary of the Invention

[0006] Object of the Invention: The first object of the present invention is to provide a 3-cyano-substituted quinoline compound, the second object is a pharmaceutical composition containing the compound (for example: a composition used in combination with antibodies such as PD-1 antibody and MCT1 antibody), and the third object is to provide the application of the compound and its pharmaceutical composition in the preparation of HPK1 inhibitor drugs.

[0007] Technical Solution: The 3-cyano-substituted quinoline compound described in the present invention has the structure of formula I, and also includes its pharmaceutically acceptable salts.

[0008]

[0009] Wherein:

[0010] R 1 is selected from C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino, C 6-10 aryl;

[0011] R 2 Selected from C 3-8 cycloalkyl, oxirane, aziridine, thiirane, oxetane, azetidine, thietane, tetrahydrofuran, pyrrolidine, tetrahydrothiophene, pyridine, pyrimidine, pyrazine, pyrrole, furan, thiophene, imidazole, pyrazole, oxazole, thiazole, 5-6 membered aromatic ring, 5-membered fused 6-membered aromatic ring, 6-membered fused 6-membered aromatic ring, quinoline, isoquinoline, indole, benzofuran, benzothiophene;

[0012] R 2 is substituted by 1 to 4 identical or different Rs;

[0013] The said R is selected from a hydrogen atom, halogen, hydroxyl, cyano, nitro, carboxyl, -NH 2 , C 1 -C 6 sulfonyl, nitro, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino; The C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino is substituted by 1 or 2 or 3 substituents, and the said substituents are selected from halogen, hydroxyl, cyano, C 1 -C 6 alkyl, C 3 -C 7 cycloalkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, phenyl;

[0014] R 3 is selected from hydrogen, deuterium, C 1 -C 6 alkyl, C 1 -C 6 acyl, substituted benzene ring, 5-6 membered aromatic ring, saturated or unsaturated 3- to 7-membered cyclic alkyl, aziridine, pyrrole, pyrrolidine, piperidine, morpholine, thiomorpholine, 1,1-dioxothiomorpholine, piperazine, cyclohexylimine, trifluoromethyl, or R 3 does not exist;

[0015] X is selected from an oxygen atom, NH, a sulfur atom;

[0016] Y is selected from CH 2, oxygen atom, nitrogen atom, sulfur atom.

[0017] Preferably, in the structure:

[0018] R 1 is selected from C 1 -C 6 alkyl, C 1 -C 6 alkylamino;

[0019] R 2 is selected from 5-6 membered aromatic rings, tetrahydrofuran, tetrahydropyrrole, tetrahydrothiophene, pyridine, pyrimidine, pyrrole, furan, thiophene, 5-6 membered aromatic rings, 5-membered fused 6-membered aromatic rings, 6-membered fused 6-membered aromatic rings, quinoline, isoquinoline, indole, benzofuran, benzothiophene;

[0020] R 2 is substituted by 1-4 identical or different Rs;

[0021] R is selected from hydrogen atom, halogen, nitro, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino.

[0022] Preferably, in the structure:

[0023] R 3 is selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 acyl, saturated or unsaturated three- to seven-membered cyclic alkyl, aziridine, pyrrole, tetrahydropyrrole, trifluoromethyl, or R 3 does not exist;

[0024] X is selected from oxygen atom, sulfur atom;

[0025] Y is selected from CH 2 , oxygen atom, nitrogen atom;

[0026] Preferably, in the structure:

[0027] R 1 is selected from C 1 -C 6 alkyl;

[0028] R 2 is preferably 5-6 membered aromatic rings, pyridine, pyrimidine, pyrrole, quinoline, isoquinoline, indole, benzofuran, benzothiophene;

[0029] R 2substituted by 1 to 4 identical or different Rs;

[0030] R is selected from halogen, nitro, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino.

[0031] More preferably, in the structure:

[0032] R 1 is selected from C 1 -C 4 alkyl;

[0033] R 2 is selected from 5- to 6-membered aromatic rings, pyridine, pyrimidine, quinoline, isoquinoline, pyrrole, indole;

[0034] R 2 is substituted by 1 to 4 identical or different Rs;

[0035] R is selected from a hydrogen atom, halogen, nitro, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylamino;

[0036] R 3 is selected from hydrogen, C 1 -C 6 alkyl, C 1 -C 6 acyl, saturated or unsaturated 3- to 7-membered cyclic alkyl, aziridine, pyrrole, pyrrolidine, trifluoromethyl, or R 3 is absent;

[0037] X is selected from an oxygen atom, and Y is selected from a nitrogen atom and an oxygen atom.

[0038] Even more preferably, R 1 is selected from methyl, ethyl;

[0039] R 2 is selected from

[0040] R 3 is selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, or R 3 is absent.

[0041] Most preferably, the 3-cyano-substituted quinoline compounds of the present invention are selected from the compounds of any of the following:

[0042]

[0043]

[0044]

[0045] The pharmaceutically acceptable salts of the 3-cyano-substituted quinoline compounds of the present invention are salts formed by the compounds and acids or bases. The acids are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, fumaric acid, succinic acid, salicylic acid, phenylacetic acid or mandelic acid, more preferably hydrochloric acid; the bases are bases containing sodium, potassium, calcium, ammonium cations, triethylamine, diethylamine or choline.

[0046] "Pharmaceutically acceptable salts" refer to salts of compounds prepared from compounds with specific substituents and relatively non-toxic acids or bases. When a compound contains relatively acidic functional groups, base addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When a compound contains relatively basic functional groups, acid addition salts can be obtained by contacting the free form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, and the inorganic acids include, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonates or bicarbonates), phosphoric acid (forming phosphates, monohydrogen phosphates, dihydrogen phosphates), sulfuric acid (forming sulfates or bisulfates), hydroiodic acid, phosphorous acid, etc.; and organic acid salts, and the organic acids include, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid and similar acids; organic acid salts also include salts of amino acids (such as arginine, etc.), glucuronic acid and other organic acids. When certain specific compounds contain both basic and acidic functional groups, they can thus be converted into either base or acid addition salts. Preferably, the salt is contacted with a base or acid in a conventional manner, and then the parent compound is separated to regenerate the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents.

[0047] "Pharmaceutically acceptable salts" can be synthesized from parent compounds containing acid or base groups by conventional chemical methods. In general, the preparation of such salts involves reacting these compounds in the form of the free acid or base with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both. Generally, non-aqueous media such as ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

[0048] The compounds described in the present invention can also be used as solvates.

[0049] The present invention includes various deuterated forms of the compounds described. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom.

[0050] In addition, the present invention also includes prodrugs of the compounds described. The prodrugs are derivatives of general formula I, which may themselves have weak or even no activity, but are converted into the corresponding bioactive forms under physiological conditions (e.g., by metabolism, solvolysis, or otherwise) after administration. The compounds shown in general formula I can exist in non-solvated form and solvated form containing pharmaceutically acceptable solvents (such as water, ethanol, etc.); the compounds shown in general formula I can contain asymmetric or chiral centers and thus can exist in different stereoisomeric forms; all stereoisomeric forms of the present invention, including but not limited to diastereoisomers, enantiomers, and atropisomers, as well as mixtures thereof (such as racemic mixtures), are included within the scope of the present invention.

[0051] The pharmaceutical compositions described in the present invention contain the 3-cyano-substituted quinoline compounds described in the present invention, compositions used in combination with antibodies such as PD-1 antibody and MCT1 antibody, and pharmaceutically acceptable carriers.

[0052] More preferably, the dosage forms of the pharmaceutical compositions described in the present invention are selected from capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories, patches. The carriers that can be arbitrarily mixed can be changed according to the dosage form, administration form, etc. Examples of carriers include excipients, binders, disintegrants, lubricants, flavoring agents, fragrances, coloring agents, or sweetening agents, etc.

[0053] "Pharmaceutically acceptable carrier" may be an excipient widely used in the field of drug production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide methods to enable the active ingredient to dissolve at a desired rate after the subject receives the administration, or to promote the effective absorption of the active ingredient after the subject receives the composition administration. The described pharmaceutical excipients can be inert fillers or provide certain functions, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The described pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesives, glidants, wetting agents, gelling agents, absorption retardants, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0054] The pharmaceutical composition described in the present invention can be prepared by any method known to those skilled in the art according to the disclosed content. For example, conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilization processes.

[0055] The pharmaceutical composition described in the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular) administration. The pharmaceutical composition of the present invention can also be in a controlled-release or sustained-release dosage form (such as liposomes or microspheres). Examples of solid oral preparations include but are not limited to powders, capsules, caplets, soft capsules and tablets. Examples of liquid preparations for oral or mucosal administration include but are not limited to suspensions, emulsions, elixirs and solutions. Examples of topical preparations include but are not limited to emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum preparations. Examples of preparations for parenteral administration include but are not limited to injection solutions, dry powder preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, injection suspensions and injection emulsions. Examples of other suitable preparations of the described pharmaceutical composition include but are not limited to eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0056] The 3-cyano-substituted quinoline compound or pharmaceutical composition described in the present invention is used in the preparation of a drug as an HPK1 inhibitor.

[0057] Preferably, the drug is a drug for preventing and / or treating cancer, autoimmune diseases or immune-mediated diseases.

[0058] More preferably, the application of the pharmaceutical composition in the treatment and / or prevention of drugs for lung cancer, prostate cancer, colorectal cancer, melanoma, ovarian cancer, breast cancer, kidney cancer, lymphoma or leukemia or autoimmune diseases.

[0059] Further preferably, the pharmaceutical composition is used for the treatment and / or prevention of acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiple myeloma, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma, breast cancer, non-small cell lung cancer, melanoma, renal cancer, ovarian cancer, prostate cancer, colon cancer and central nervous system tumors, or for the treatment of autoimmune diseases.

[0060] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages:

[0061] This class has excellent HPK1 inhibitory activity (the inhibitory IC 50 value reaches the nanomolar concentration level), good water solubility and pharmacokinetic stability, has the potential to become a drug, and can be used in the preparation of drugs for the prevention and / or treatment of cancer, inflammatory diseases, autoimmune diseases or immune-mediated diseases. Detailed implementation manners

[0062] The technical solution of the present invention will be further described below in conjunction with the embodiments.

[0063] The compound structure of the present invention was confirmed by nuclear magnetic resonance hydrogen spectrum ( 1 1H-NMR) and mass spectrometry (MS). The purity of the compound was determined by high performance liquid chromatography (HPLC). 1 The measurement of 1H-NMR was carried out using Bruker Advance 300 and Bruker Advance 400 nuclear magnetic resonance spectrometers. The measurement solvents were deuterated dimethyl sulfoxide (DMSO-d 6 ) and deuterated chloroform (CDCl 3 ), and the internal standard was tetramethylsilane (TMS).

[0064] Thin layer chromatography (TLC) used thin layer chromatography silica gel plates. The self-made silica gel thin layer plates used GF254 silica gel, and silica column chromatography generally used 200-300 mesh silica gel.

[0065] The starting materials in the examples are known and can be purchased on the market, or can be synthesized according to methods known in the art.

[0066] Unless otherwise specified, all reactions were carried out under continuous magnetic stirring, the solvent was a dry solvent, and the reaction temperature unit was degrees Celsius.

[0067] Example 1: Preparation of 1-(3-cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0068]

[0069] 1-(3-Cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea of the present invention can be synthesized according to the following reaction route:

[0070]

[0071] Preparation of ethyl 3-(4-acetamido-3-ethoxyaniline)-2-cyanoacrylate (II):

[0072] Step 1: Add N-(4-amino-2-ethoxyphenyl)acetamide (I) (10 g, 51.5 mmol) and (E)-ethyl 2-cyano-3-ethoxyacrylate (9.6 g, 56.6 mmol) to toluene (50 ml), and react at 100 °C for 4 h; Detect by TLC that the raw materials disappear and the reaction is complete; After cooling the reaction solution, a large amount of pale yellow solid precipitates. Filter it by suction to obtain the crude product. Wash the crude product with toluene to obtain the pale yellow solid (II) (16.1 g, 50.8 mmol), and the yield is 85%. 1 H NMR(400MHz,Chloroform-d)δ9.37(s,1H),7.99(d,J=8.4Hz,1H),7.53(d,J=7.5Hz,1H),7.12(d,J=1.5Hz,1H),7.07(dd,J=7.5,1.5Hz,1H),4.31(q,J=8.0Hz,2H),4.12(q,J=8.0Hz,2H),2.19(s,2H),1.42(t,J=8.0Hz,3H),1.34(t,J=8.0Hz,3H);ESI-MS m / z:317.1calcd for C 16 H 19 N 3 O 4 [M+H] + 318.1.

[0073] Step 2: Preparation of 3-cyano-4-hydroxy-6-acetamido-7-ethoxyquinoline (III):

[0074] Ethyl 3-(4-acetamido-3-ethoxyaniline)-2-cyanoacrylate (II) (5 g, 15.8 mmol) was added to diphenyl ether (30 ml), and the reaction was carried out at 260 °C for 3 h. The reaction was completed as detected by TLC when the raw materials disappeared. Petroleum ether was added to diphenyl ether, and a large amount of brown solid precipitated. The crude product was obtained by suction filtration, and the crude product was washed with petroleum ether three times to obtain a brown solid (III) (4 g, 14.7 mmol), with a yield of 93%. ESI-MS m / z: 271.1 calcd for C 14 H 13 N 3 O 3 [M+H] + 272.1.

[0075] Step 3: Preparation of N-(4-chloro-3-cyano-7-ethoxy-6-quinolinyl)acetamide (IV):

[0076] 3-Cyano-4-hydroxy-6-acetamido-7-ethoxyquinoline (III) (2 g, 7.4 mmol) was added to an acetonitrile (20 ml) solution, and POCl 3 (2 ml, 22.1 mmol) was added. The reaction was carried out at 90 °C for 10 h. The reaction was completed as detected by TLC when the raw materials disappeared. The solvent was evaporated to dryness, and the residue was extracted with EA. The organic layer was washed three times with H 2 O and saturated brine, dried over anhydrous sodium sulfate, allowed to stand, filtered, and the solvent was removed by distillation under reduced pressure. The product was separated by silica gel column chromatography to obtain a brown solid (IV) (1.8 g, 6.3 mmol), with a yield of 85%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.18 (s, 1H), 8.79 (s, 1H), 8.59 (s, 1H), 7.71 (s, 1H), 4.11 (q, J = 8.0 Hz, 3H), 2.20 (s, 3H), 1.41 (t, J = 8.0 Hz, 3H); ESI-MS m / z: 289.1 calcd for C 14 H 12 ClN 3 O 2 [M+H] + 290.1.

[0077] Step 4: Preparation of N-(3-cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)acetamide (V):

[0078] N-(4-chloro-3-cyano-7-ethoxy-6-quinolinyl)acetamide (IV) (200 mg, 0.69 mmol) and p-toluidine (81 mg, 0.76 mmol) were added to a solution of DMF (10 ml). The mixture was stirred for 15 min in an ice bath, and sodium bis(trimethylsilyl)amide (1.0 M solution in THF) (2 ml) was added. The reaction was carried out for 2 h in an ice bath. The reaction was completed as detected by TLC when the starting materials disappeared. The mixture was extracted with EA, and the organic layer was washed three times with H 2 O and saturated brine, dried over anhydrous sodium sulfate, allowed to stand, filtered, and the solvent was removed by distillation under reduced pressure. The residue was purified by trituration with ethyl acetate and petroleum ether to give a white solid (V) (240 mg, 0.67 mmol), with a yield of 97%. 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.46 (s, 1H), 8.59 (s, 1H), 8.15 (s, 1H), 7.82 (s, 1H), 7.56 (s, 1H), 7.14 (dt, J = 6.9, 0.8 Hz, 3H), 7.6 (m, 3H), 4.11 (q, J = 8.0 Hz, 2H), 2.33 (s, J = 0.9 Hz, 3H), 2.15 (s, 3H), 1.40 (m, 3H); ESI-MS m / z: 360.2 calcd for C 21 H 20 N 4 O 2 [M + H] + 361.2.

[0079] Step 5: Preparation of 6-amino-7-ethoxy-4-(p-benzylamino)quinoline-3-carbonitrile (VI):

[0080] N-(3-Cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)acetamide (V) (200 mg, 0.55 mmol) was added to 2N HCl solution. The reaction was carried out at 100 °C for 24 h. The reaction was completed as detected by TLC when the starting materials disappeared. The pH value was adjusted to 12 with 2N NaOH solution, and the mixture was extracted with EA. The organic layer was washed three times with H 2 O and saturated brine, dried over anhydrous sodium sulfate, allowed to stand, filtered, and the solvent was removed by distillation under reduced pressure. The residue was separated by silica gel column chromatography to give a yellow solid (VI) (133 mg, 0.42 mmol), with a yield of 76%. 1 1H NMR (400 MHz, DMSO-d 6)δ8.54(s,1H),8.28(s,1H),7.76(s,1H),7.24(s,1H),7.16(d,J=0.9Hz,1H),7.14(d,J=0.8Hz,1H),7.09(s,1H),7.08(d,J=1.4Hz,1H),4.90(dd,J=44.0,6.9Hz,2H),4.07(q,J=8.0Hz,2H),2.33(d,J=0.9Hz,3H),1.42(t,J=8.0Hz,3H); ESI-MS m / z: 318.1 calcd for C 19 H 18 N 4 O[M+H] + 319.1.

[0081] Step 6: Preparation of 1-(3-cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (VII):

[0082] 6-Amino-7-ethoxy-4-(p-benzylamino)quinoline-3-carbonitrile (VI) (100 mg, 0.31 mmol) was added to a solution of tetrahydrofuran. After stirring for 15 min in an ice bath, DIPEA (161 μl, 0.93 mmol) and triphosgene (34 mg, 0.10 mmol) were added. After stirring for 30 min, 1-ethyl-4-aminopiperidine (40 mg, 0.31 mmol) was added, and the reaction was carried out at 0 °C for 3 h. The reaction was monitored by TLC until the raw materials disappeared. The pH value was adjusted to 12 with 2N NaOH solution, and the mixture was extracted with EA. The organic layer was washed three times with H 2 2O and saturated brine, dried over anhydrous sodium sulfate, allowed to stand, filtered, and the solvent was removed by distillation under reduced pressure. The product was separated by silica gel column chromatography to obtain a white solid (VII) (98 mg, 0.21 mmol), with a yield of 68%. 1 1H NMR (400 MHz, DMSO-d 6 ) δ8.76(s,1H),8.61(s,1H),8.39(s,1H),7.73(s,1H),7.35(s,1H),7.16(m,2H),7.10(m,2H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.67(dp,J=10.3,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.55(q,J=8.0Hz,2H),2.32(d,J=0.6Hz,3H),1.80(m,4H),1.41(t,J=8.0Hz,3H),1.05(t,J=8.0Hz,3H).13 13C NMR (75 MHz, DMSO-d 6 ) δ 157.63, 152.88, 149.11, 146.18, 142.60, 140.23, 129.50, 129.39, 128.34, 119.62, 114.98, 114.15, 112.63, 105.55, 90.74, 65.16, 51.47, 50.56, 47.87, 31.27, 20.71, 14.93, 12.20; ESI-MS m / z: 472.3 calcd for C 27 H 32 N 6 O 2 [M + H] + 473.3.

[0083] Example 2: Preparation of 1-(3-Cyano-7-ethoxy-4-((4-nitrophenyl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0084]

[0085] In this example, p-toluidine was replaced with p-nitroaniline, and according to the operation of Example 1, 78 mg of the product was obtained with a yield of 54%; 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.97 (s, 1H), 8.76 (s, 1H), 8.60 (s, 1H), 8.40 (s, 1H), 8.10 (m, 2H), 7.40 (m, 3H), 6.03 (d, J = 10.3 Hz, 1H), 4.10 (q, J = 8.0 Hz, 2H), 3.66 (dp, J = 10.3, 7.0 Hz, 1H), 2.86 (qt, J = 12.5, 7.1 Hz, 4H), 2.54 (q, J = 8.1 Hz, 2H), 1.82 (m, 4H), 1.41 (t, J = 8.0 Hz, 3H), 1.06 (m, 3H); 13 13C NMR (75 MHz, DMSO-d 6 ) δ 157.63, 152.88, 149.11, 146.18, 144.10, 142.80, 142.22, 129.39, 125.45, 118.56, 114.98, 113.93, 112.63, 105.57, 90.74, 65.05, 51.42, 50.56, 47.74, 30.86, 14.93, 12.20; ESI-MS m / z: 503.2 calcd for C 26 H 29N 7 O 4 [M+H] + 504.2

[0086] Example 3: Preparation of 1-(4-((4-chloro-2-fluorophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0087]

[0088] In this example, p-toluidine was replaced with 4-chloro-2-fluoroaniline. According to the operation of Example 1, 96 mg of the product was obtained, and the yield was 67%; 1 H NMR(400MHz, DMSO-d 6 )δ8.76(s,1H),8.57(d,J = 2.5Hz,2H),8.39(s,1H),7.39(dd,J = 7.5,4.9Hz,1H),7.36(s,1H),7.23(dd,J = 8.0,1.5Hz,1H),7.14(dd,J = 7.5,1.5Hz,1H),6.03(d,J = 10.3Hz,1H),4.10(q,J = 8.0Hz,2H),3.67(dp,J = 10.3,7.0Hz,1H),2.86(qt,J = 12.5,7.1Hz,4H),2.54(q,J = 8.0Hz,2H),1.77(m,4H),1.41(t,J = 8.0Hz,3H),1.06(m,3H); 13 C NMR(75MHz, DMSO-d 6 )δ157.63,154.32,152.31,151.64,149.11,146.46,143.55,143.45,131.91,131.85,131.83,131.67,129.39,125.85,125.82,122.32,122.26,116.23,116.07,114.98,114.15,112.63,105.57,90.60,65.05,51.42,50.56,47.74,30.86,14.93,12.20; ESI-MS m / z: 510.2 calcd for C 26 H 28 ClFN 6 O 2 [M+H] + 511.2

[0089] Example 4: Preparation of 1-(3-cyano-4-((2,4-difluoro-5-methoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0090]

[0091] In this example, p-toluidine was replaced with 2,4-difluoro-5-methoxyaniline. According to the operation of Example 1, 85 mg of the product was obtained with a yield of 60%; 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.76 (s, 1H), 8.67 (s, 1H), 8.60 (s, 1H), 8.37 (s, 1H), 7.39 (s, 1H), 7.02 (t, J = 8.0 Hz, 1H), 6.76 (t, J = 5.0 Hz, 1H), 6.03 (d, J = 10.3 Hz, 1H), 4.10 (q, J = 8.0 Hz, 2H), 3.88 (s, 2H), 3.66 (dp, J = 10.2, 7.0 Hz, 1H), 2.86 (qt, J = 12.5, 7.1 Hz, 4H), 2.54 (q, J = 8.0 Hz, 2H), 1.76 (qq, J = 12.5, 7.0 Hz, 4H), 1.41 (t, J = 8.0 Hz, 3H), 1.05 (t, J = 8.0 Hz, 3H); 13 C NMR (75 MHz, DMSO-d 6 ) δ 157.63, 151.66, 149.50, 149.11, 148.94, 148.87, 147.48, 146.46, 146.06, 145.90, 143.30, 129.78, 129.75, 129.39, 114.98, 113.96, 112.63, 106.80, 106.74, 105.57, 103.94, 103.77, 90.60, 64.93, 56.58, 56.55, 51.42, 50.29, 47.74, 30.78, 14.93, 12.20; ESI-MS m / z: 524.2 calcd for C 27 H 30 F 2 N 6 O 3 [M + H] + 525.2.

[0092] Example 5: Preparation of 1-(3-cyano-7-ethoxy-4-((2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0093]

[0094] In this example, p-toluidine was replaced with 1,2,3,4-tetrahydro-2-methyl-7-isoquinolineamine. According to the operation of Example 1, 80 mg of the product was obtained with a yield of 56%; 1 H NMR(400MHz, DMSO-d 6 ) δ8.76(s, 1H), 8.59(s, 1H), 8.37(s, 1H), 8.07(s, 1H), 7.40(s, 1H), 7.10(dt, J = 7.4, 1.0Hz, 1H), 6.99(dd, J = 7.5, 1.6Hz, 1H), 6.76(q, J = 1.0Hz, 1H), 6.03(d, J = 10.3Hz, 1H), 4.10(q, J = 8.0Hz, 2H), 3.65(m, 3H), 2.96(t, J = 7.1Hz, 2H), 2.86(qt, J = 12.5, 7.1Hz, 4H), 2.75(m, 2H), 2.54(q, J = 8.0Hz, 2H), 2.44(s, 2H), 1.76(qq, J = 12.5, 7.1Hz, 4H), 1.40(m, 3H), 1.05(m, 3H); 13 C NMR(75MHz, DMSO-d 6 ) δ157.63, 151.76, 149.11, 146.18, 142.56, 139.13, 131.07, 130.48, 129.49, 127.32, 119.32, 119.10, 115.00, 114.38, 112.75, 105.57, 90.74, 64.77, 56.57, 51.42, 51.33, 50.29, 47.74, 43.86, 30.78, 29.10, 14.93, 12.20; ESI-MS m / z: 527.3 calcd for C 30 H 37 N 7 O 2 [M + H] + 528.3.

[0095] Example 6: Preparation of 1-(3-cyano-7-ethoxy-4-(phenylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0096]

[0097] In this example, p-toluidine was replaced with aniline. According to the operation of Example 1, 80 mg of the product was obtained with a yield of 80%; 1HNMR(400MHz, DMSO-d 6 ) δ 8.76 (s, 1H), 8.61 (s, 1H), 8.40 (s, 1H), 7.97 (s, 1H), 7.38 (s, 1H), 7.23 (m, 2H), 7.15–7.09 (m, 2H), 6.94 (tt, J = 7.3, 1.5 Hz, 1H), 6.03 (d, J = 10.3 Hz, 1H), 4.10 (q, J = 8.0 Hz, 2H), 3.67 (dp, J = 10.3, 7.0 Hz, 1H), 2.85 (m, 4H), 2.55 (q, J = 8.0 Hz, 2H), 1.76 (m, 4H), 1.41 (t, J = 8.0 Hz, 3H), 1.05 (t, J = 8.0 Hz, 3H); 13 C NMR(75MHz, DMSO-d 6 ) δ 157.63, 152.85, 149.11, 146.41, 142.60, 141.55, 129.39, 128.81, 122.64, 119.76, 115.03, 114.15, 112.55, 105.61, 90.74, 65.16, 51.47, 50.57, 47.74, 31.27, 14.93, 12.20; ESI-MS m / z: 458.3 calcd for C 26 H 30 N 6 O 2 [M + H] + 459.3.

[0098] Example 7: Preparation of 1-(3-cyano-7-ethoxy-4-((1-methyl-1H-indol-6-yl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0099]

[0100] In this example, p-toluidine was replaced with 1-methyl-6-aminoindole, and according to the operation of Example 1, 125 mg of the product was obtained with a yield of 87%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.40(s,1H),7.75(m,1H),7.69(s,1H),7.40(s,1H),7.26(dt,J=7.5,0.7Hz,1H),7.12(d,J=1.8Hz,1H),7.04(dd,J=7.4,1.5Hz,1H),6.48(m,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.75(d,J=0.7Hz,3H),3.66(dp,J=10.4,7.1Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.77(qd,J=7.1,4.1Hz,4H),1.41(t,J=8.0Hz,3H),1.05(m,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.76,149.11,146.18,142.68,138.93,136.85,129.39,129.08,125.04,119.90,114.98,114.55,114.06,112.63,105.57,103.15,96.62,90.74,65.05,51.42,50.56,47.74,32.94,30.86,14.93,12.20;ESI-MS m / z:511.3calcd forC 29 H 33 N 7 O 2 [M+H] + 512.3.

[0101] Example 8: Preparation of 1-(3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0102]

[0103] In this example, p-toluidine was replaced with 2,4-dichloro-5-methoxyaniline. According to the operation of Example 1, 114 mg of the product was obtained with a yield of 82%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.39(s,1H),8.35(s,1H),7.48(s,1H),7.39(s,1H),6.83(s,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.88(s,2H),3.66(dp,J=10.2,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.83–1.67(m,4H),1.40(t,J=8.0Hz,3H),1.05(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,155.56,151.64,149.11,146.39,142.38,139.76,130.03,129.39,122.37,115.45,114.98,114.38,112.75,105.56,105.55,90.60,64.51,56.37,51.36,50.29,47.74,30.78,14.93,12.20;ESI-MS m / z:556.2calcd for C 27 H 30 Cl 2 N 6 O 2 [M+H] + 557.2.

[0104] Example 9: Preparation of 1-(3-cyano-4-((2,4-dichlorophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0105]

[0106] In this example, p-toluidine was replaced with 2,4-dichloroaniline, and according to the operation of Example 1, 65 mg of the product was obtained with a yield of 46%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.54(s,1H),8.38(s,1H),8.06(s,1H),7.49(d,J=1.5Hz,1H),7.39(s,1H),7.34(dd,J=7.5,1.5Hz,1H),7.28(d,J=7.5Hz,1H),6.03(d,J=10.2Hz,1H),4.10(q,J=8.0Hz,2H),3.67(dp,J=10.2,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.82(dq,J=12.4,7.1Hz,2H),1.72(dq,J=12.3,7.0Hz,2H),1.41(t,J=8.0Hz,3H),1.05(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.64,149.11,146.24,142.53,138.29,130.83,129.51,129.39,128.48,127.45,122.92,114.98,114.04,112.63,105.57,90.60,64.93,51.42,50.29,47.74,30.78,14.93,12.20;ESI-MS m / z:526.2calcd for C 26 H 28 Cl 2 N 6 O 2 [M+H] + 527.2.

[0107] Example 10: Preparation of 1-(4-((1H-indazol-5-yl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0108]

[0109] In this example, p-toluidine was replaced with 6-aminoindazole, and according to the operation of Example 1, 88 mg of the product was obtained with a yield of 61%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.41(d,J=12.1Hz,2H),8.09(d,J=1.6Hz,1H),7.61(t,J=1.6Hz,1H),7.49(d,J=7.3Hz,1H),7.39(s,1H),7.31(dd,J=7.5,1.5Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.66(dp,J=10.3,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.1Hz,2H),1.76(p,J=7.1Hz,4H),1.41(t,J=8.0Hz,3H),1.05(m,3H); 13 CNMR(75MHz,DMSO-d 6 )δ157.63,152.90,149.11,146.18,142.56,137.49,134.95,131.96,129.39,124.21,119.82,114.98,113.93,112.63,111.81,111.59,105.57,90.74,65.05,51.47,50.56,47.74,30.86,14.93,12.20;ESI-MS m / z:498.2calcd for C 27 H 30 N 8 O 2 [M+H] + 499.2.

[0110] Example 11: Preparation of 1-(4-((2-chloro-5-nitrophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0111]

[0112] In this example, p-toluidine was replaced with 2-chloro-5-nitroaniline, and according to the operation of Example 1, 113 mg of the product was obtained with a yield of 71%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.45–8.40(m,2H),8.37(s,1H),7.97(dd,J=7.5,1.6Hz,1H),7.62(d,J=7.4Hz,1H),7.35(s,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.66(dp,J=10.2,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.83–1.67(m,4H),1.40(t,J=8.0Hz,3H),1.05(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.64,149.11,147.75,146.39,142.49,139.46,131.64,129.39,128.14,118.69,115.43,114.98,113.62,112.63,105.57,90.60,64.93,51.36,50.29,47.74,30.78,14.93,12.20;ESI-MS m / z:537.2calcd forC 26 H 28 ClN 7 O 4 [M+H] + 538.2.

[0113] Example 12: Preparation of 1-(4-((2-chloro-4-fluorophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0114]

[0115] In this example, p-toluidine was replaced with 2-chloro-4-fluoroaniline, and according to the operation of Example 1, 78 mg of the product was obtained with a yield of 54%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.53(s,1H),8.39(s,1H),8.03(s,1H),7.43–7.37(m,2H),7.23(dd,J=8.1,1.6Hz,1H),7.02(td,J=7.8,1.6Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.67(dp,J=10.3,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.87–1.70(m,4H),1.41(t,J=8.0Hz,3H),1.09–1.02(m,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.64,149.11,146.24,142.58,129.39,122.87,122.80,117.23,117.07,116.84,116.68,114.98,114.04,112.63,105.57,90.60,65.05,51.42,50.56,47.74,30.86,14.93,12.20;ESI-MS m / z:510.2calcd forC 26 H 28 ClFN 6 O 2 [M+H] + 511.2.

[0116] Example 13: Preparation of 1-(4-((2-chloro-5-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea

[0117]

[0118] In this example, p-toluidine was replaced with 2-chloro-5-methoxyaniline, and 1-ethyl-4-aminopiperidine was replaced with 1-isopropyl-4-piperidinamine. According to the operation of Example 1, 56 mg of the product was obtained with a yield of 39%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.37(s,1H),8.22(s,1H),7.39(s,1H),7.29(d,J=7.4Hz,1H),6.90(d,J=1.5Hz,1H),6.69(dd,J=7.5,1.6Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.80(s,2H),3.67(dp,J=10.4,7.0Hz,1H),2.88–2.76(m,3H),2.70(dt,J=12.5,7.1Hz,2H),1.87–1.69(m,4H),1.44–1.37(m,3H),1.09(d,J=6.8Hz,5H); 13 C NMR(75MHz,DMSO-d 6 )δ158.72,157.63,151.64,149.11,146.39,142.43,140.13,131.12,129.39,120.99,114.98,114.04,112.63,110.08,107.06,105.57,90.60,64.93,55.47,54.22,47.95,47.78,31.04,18.70,14.93;ESI-MS m / z:536.2calcd for C 28 H 33 ClN 6 O 3 [M+H] + 537.2.

[0119] Example 14: Preparation of 1-(3-cyano-4-((2,4-dichlorophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-cyclopropylpiperidin-4-yl)urea

[0120]

[0121] In this example, p-toluidine was replaced with 2-chloro-5-methoxyaniline, and 1-ethyl-4-aminopiperidine was replaced with 1-isopropyl-4-piperidinamine. According to the operation of Example 1, 72 mg of the product was obtained with a yield of 50%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.54(s,1H),8.37(s,1H),8.06(s,1H),7.49(d,J=1.5Hz,1H),7.39(s,1H),7.37–7.29(m,2H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.66(dp,J=10.3,7.0Hz,1H),2.81(dt,J=12.4,7.1Hz,2H),2.69(dt,J=12.4,7.1Hz,2H),2.36(p,J=6.9Hz,1H),1.87–1.72(m,4H),1.46–1.33(m,5H),1.27–1.19(m,1H),1.23–1.16(m,1H); 13 CNMR(75MHz,DMSO-d 6 )δ157.63,152.18,149.11,146.39,142.53,138.29,130.83,129.52,129.39,128.38,128.24,122.92,114.98,114.02,112.63,105.57,90.60,64.93,48.59,47.78,35.87,31.01,14.93,8.39;ESI-MS m / z:538.2calcd for C 27 H 28 Cl 2 N 6 O 2 [M+H] + 539.2.

[0122] Example 15: Preparation of 1-(4-((4-chloro-3-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-methylpiperidin-4-yl)urea

[0123]

[0124] In this example, p-toluidine was replaced with 3-methoxy-4-chloroaniline, and 1-ethyl-4-aminopiperidine was replaced with 4-amino-1-methylpiperidine. According to the operation of Example 1, 120 mg of the product was obtained with a yield of 87%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.39(s,1H),7.95(s,1H),7.40(s,1H),7.34(d,J=7.5Hz,1H),6.86(dd,J=7.5,1.5Hz,1H),6.83(d,J=1.4Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.89(s,2H),3.67(dp,J=10.2,7.0Hz,1H),2.83(dt,J=12.5,7.1Hz,2H),2.49(dt,J=12.5,7.1Hz,2H),2.35(s,2H),1.79(qd,J=7.1,1.2Hz,4H),1.41(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,155.99,152.31,149.11,146.18,142.58,141.26,131.05,129.39,119.64,114.98,114.53,114.15,112.63,105.87,105.57,90.74,65.05,56.78,55.06,47.67,46.01,30.88,14.93;ESI-MS m / z:508.2calcd for C 26 H 29 ClN 6 O 3 [M+H] + 509.2.

[0125] Example 16: Preparation of 1-(4-((4-chloro-3-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea

[0126]

[0127] In this example, p-toluidine was replaced with 3-methoxy-4-chloroaniline, and 1-ethyl-4-aminopiperidine was replaced with 4-aminotetrahydropyran. According to the operation of Example 1, 105 mg of the product was obtained with a yield of 78%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.40(s,1H),8.19(s,1H),7.39(s,1H),7.32(d,J=7.5Hz,1H),6.86(dd,J=7.5,1.5Hz,1H),6.83(d,J=1.4Hz,1H),5.90(d,J=10.4Hz,1H),4.10(q,J=8.0Hz,2H),3.89(s,2H),3.75–3.59(m,6H),1.99–1.82(m,4H),1.41(t,J=8.0Hz,3H); 13 CNMR(75MHz,DMSO-d 6 )δ157.24,155.99,152.88,149.11,146.18,142.53,141.26,131.05,129.39,119.64,114.98,114.53,114.03,112.63,105.87,105.57,90.74,66.71,65.09,56.78,46.77,32.01,14.93;ESI-MS m / z:495.2calcd for C 25 H 26 ClN 5 O 4 [M+H] + 496.2.

[0128] Example 17: Preparation of 1-(4-((4-bromo-2-methylphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea

[0129]

[0130] In this example, p-toluidine was replaced with 2-methyl-4-bromoaniline, and 1-ethyl-4-aminopiperidine was replaced with 4-aminotetrahydropyran. According to the operation of Example 1, 70 mg of the product was obtained with a yield of 53%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.53(s,1H),8.41(s,1H),8.20(s,1H),7.40(s,1H),7.36(d,J=1.4Hz,1H),7.31(dd,J=7.5,1.5Hz,1H),7.10(d,J=7.5Hz,1H),5.90(d,J=10.4Hz,1H),4.10(q,J=8.0Hz,2H),3.75–3.58(m,6H),2.25(s,2H),1.99–1.82(m,4H),1.41(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.24,152.10,149.11,146.47,142.63,138.90,132.80,130.13,129.62,129.39,121.77,116.53,114.98,114.19,112.63,105.57,90.60,66.86,65.03,46.92,32.07,17.89,14.93;ESI-MS m / z:523.1calcd for C 25 H 26 BrN 5 O 3 [M+H] + 524.1.

[0131] Example 18: Preparation of 1-(3-cyano-4-((2,4-diiodophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea

[0132]

[0133] In this example, p-toluidine was replaced with 2,4-diiodoaniline, and 1-ethyl-4-aminopiperidine was replaced with tetrahydrothiophen-4-amine. According to the operation of Example 1, 83 mg of the product was obtained with a yield of 66%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.56(s,1H),8.39(s,1H),8.30(s,1H),7.91(d,J=1.6Hz,1H),7.64(dd,J=7.5,1.5Hz,1H),7.39(s,1H),7.12(d,J=7.5Hz,1H),6.02(d,J=10.1Hz,1H),4.10(q,J=8.0Hz,2H),3.74(dp,J=10.1,7.0Hz,1H),2.70–2.58(m,4H),1.87(dq,J=12.3,7.0Hz,2H),1.69(dq,J=12.3,7.0Hz,2H),1.40(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.24,151.23,148.73,148.06,146.57,144.88,141.65,138.64,128.80,121.13,118.81,115.12,114.12,112.70,105.05,91.30,89.91,64.68,48.86,31.14,27.93,14.95;ESI-MS m / z:699.0calcdfor C 24 H 23 I 2 N 5 O 2 S[M+H] + 700.0.

[0134] Example19:Preparation of 1-(3-cyano-4-((4,5-dimethyl-2-nitrophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea

[0135]

[0136] In this example,p-toluidine was replaced with 4,5-dimethyl-2-nitroaniline,and 1-ethyl-4-aminopiperidine was replaced with tetrahydrothiophen-4-amine.According to the operation of Example 1,130 mg of the product was obtained with a yield of 94%; 1 H NMR(400MHz,DMSO-d 6)δ9.65(s,1H),8.76(s,1H),8.60(s,1H),8.36(s,1H),7.88(s,1H),7.39(s,1H),7.22(d,J=0.6Hz,1H),6.02(d,J=10.1Hz,1H),4.10(q,J=8.0Hz,2H),3.75(dp,J=10.0,7.0Hz,1H),2.70–2.58(m,4H),2.55(s,2H),2.23(s,2H),1.87(dq,J=12.5,7.0Hz,2H),1.70(dq,J=12.5,7.0Hz,2H),1.41(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.44,149.11,146.46,141.95,140.40,137.54,132.85,132.80,129.39,125.67,120.09,114.98,113.71,112.63,105.57,90.60,64.93,47.86,32.94,28.07,20.22,20.18,14.93;ESI-MSm / z:520.2calcd for C 26 H 28 N 6 O 4 S[M+H] + 521.2.

[0137] Example 20: Preparation of 1-(3-cyano-4-((3,4-dimethoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea

[0138]

[0139] In this example, p-toluidine was replaced with 3,4-dimethoxyaniline, and 1-ethyl-4-aminopiperidine was replaced with tetrahydrothiopyran-4-amine. According to the operation of Example 1, 97 mg of the product was obtained, with a yield of 69%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.40(s,1H),8.36(s,1H),7.44–7.38(m,2H),7.01(dd,J=7.4,1.5Hz,1H),6.85(d,J=7.5Hz,1H),6.02(d,J=10.1Hz,1H),4.10(q,J=8.0Hz,2H),3.82(d,J=8.2Hz,6H),3.75(dp,J=10.0,7.0Hz,1H),2.70–2.58(m,4H),1.89(dq,J=12.4,7.0Hz,2H),1.71(dq,J=12.3,7.0Hz,2H),1.41(t,J=8.0Hz,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,152.31,149.71,149.11,146.47,146.18,142.58,137.67,129.39,116.81,114.98,114.15,113.34,112.63,106.19,105.57,90.74,65.05,56.14,55.92,47.86,32.94,28.07,14.93;ESI-MS m / z:507.2calcd for C 26 H 29 N 5 O 4 S[M+H] + 508.2.

[0140] Example 21: Preparation of 1-(3-cyano-7-methoxy-4-((4-methoxy-3-(trifluoromethyl)phenyl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0141]

[0142] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzyl methyl ether, and p-toluidine was replaced with 4-methoxy-3-trifluoromethylaniline. According to the operation of Example 1, 85 mg of the product was obtained with a yield of 61%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.59(s,1H),8.40(d,J=14.6Hz,1H),7.40(s,1H),7.30(d,J=1.5Hz,1H),7.18(dd,J=7.5,1.5Hz,1H),7.08(d,J=7.5Hz,1H),6.03(d,J=10.3Hz,1H),3.95(s,2H),3.91(s,2H),3.65(dp,J=10.2,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.83–1.67(m,4H),1.08–1.01(m,2H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,152.92,152.89,152.31,150.23,146.35,142.63,137.58,137.56,129.31,128.44,126.29,121.85,119.60,119.34,117.39,117.36,115.00,114.38,114.35,114.33,112.98,104.80,90.74,56.15,55.77,51.36,50.29,47.74,30.78,12.20;ESI-MSm / z:542.2calcd for C 27 H 29 F 3 N 6 O 3 [M+H] + 543.2.

[0143] Example 22: Preparation of 1-(4-((6-bromonaphthalen-2-yl)amino)-3-cyano-7-methoxyquinolin-6-yl)-3-(1-methylpiperidin-4-yl)urea

[0144]

[0145] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzyl ether, p-toluidine was replaced with 6-bromo-2-aminonaphthalene, and 1-ethyl-4-aminopiperidine was replaced with 4-amino-1-methylpiperidine. According to the operation of Example 1, 88 mg of the product was obtained with a yield of 66%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.40(s,1H),8.09(t,J=1.6Hz,1H),8.02(s,1H),7.82(dd,J=7.6,1.5Hz,1H),7.79–7.73(m,1H),7.62(dd,J=7.6,1.6Hz,1H),7.51(p,J=0.9Hz,1H),7.34(s,1H),7.29(dd,J=7.6,1.5Hz,1H),6.03(d,J=10.3Hz,1H),3.95(s,2H),3.66(dp,J=10.2,7.0Hz,1H),2.83(dt,J=12.5,7.1Hz,2H),2.49(dt,J=12.5,7.1Hz,2H),2.35(s,2H),1.77(qd,J=7.1,3.0Hz,4H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,152.62,150.01,146.35,142.42,138.63,133.37,130.88,129.81,129.42,129.31,127.48,127.31,119.60,118.20,115.37,114.98,114.65,112.87,104.88,90.74,56.15,54.85,47.67,45.51,30.98;ESI-MS m / z:542.2calcd for C 27 H 29 F 3 N 6 O 3 [M+H] + 543.2.

[0146] Example 23: Preparation of 1-(4-((2-chloro-5-methoxyphenyl)amino)-3-cyano-7-methoxyquinolin-6-yl)-3-cyclohexylurea

[0147]

[0148] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzyl ether, p-toluidine was replaced with 2-chloro-5-methoxyaniline, and 1-ethyl-4-aminopiperidine was replaced with cyclohexylamine. According to the operation of Example 1, 67 mg of the product was obtained with a yield of 50%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.62(s,1H),8.36(s,1H),8.20(s,1H),7.33(s,1H),7.26(d,J=7.4Hz,1H),6.89(d,J=1.5Hz,1H),6.69(dd,J=7.5,1.5Hz,1H),6.00(d,J=10.3Hz,1H),3.97(s,3H),3.81(s,3H),3.41(dp,J=10.4,7.0Hz,1H),1.57–1.31(m,13H); 13 C NMR(75MHz,DMSO-d 6 )δ158.74,157.63,152.10,150.23,146.50,142.43,140.00,131.08,129.40,120.94,114.98,114.04,112.75,109.59,106.74,104.96,90.60,56.15,55.47,48.64,32.33,25.37,24.91;ESI-MS m / z:479.2calcd forC 27 H 29 F 3 N 6 O 3 [M+H] + 480.2.

[0149] Example 24: Preparation of 1-(3-cyano-7-methoxy-4-((4-methoxy-3-methylphenyl)amino)quinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea

[0150]

[0151] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzene methylether, p-toluidine was replaced with 4-methoxy-3-methylaniline, and 1-ethyl-4-aminopiperidine was replaced with 4-aminotetrahydropyran. According to the operation of Example 1, 110 mg of the product was obtained with a yield of 80%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.61(s,1H),8.57(s,1H),8.42(s,1H),7.70–7.66(m,1H),7.34(s,1H),7.10(dd,J=7.5,1.5Hz,1H),6.80(d,J=7.5Hz,1H),5.90(d,J=10.4Hz,1H),3.97(s,3H),3.82(s,3H),3.75–3.59(m,6H),2.19(s,3H),1.99–1.82(m,5H); 13 C NMR(75MHz,DMSO-d 6 )δ157.24,153.14,152.88,150.39,146.18,142.47,137.47,129.40,128.98,122.86,118.84,115.03,114.15,112.68,111.87,104.96,90.74,66.65,56.15,55.66,46.78,32.07,15.96;ESI-MS m / z:479.2calcdfor C 27 H 29 F 3 N 6 O 3 [M+H] + 480.2.

[0152] Example 25: Preparation of 1-(3-cyano-4-((2,3-dihydrobenzofuran-5-yl)amino)-7-methoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0153]

[0154] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzyl ether, and p-toluidine was replaced with 5-amino-2,3-dihydrobenzofuran[b]. According to the operation of Example 1, 90 mg of the product was obtained with a yield of 68%; 1 H NMR(400MHz,DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.38(s,1H),8.21(s,1H),7.37(s,1H),7.14(dd,J=7.4,1.6Hz,1H),6.87(q,J=1.2Hz,1H),6.84(d,J=7.5Hz,1H),6.03(d,J=10.3Hz,1H),4.52(t,J=7.1Hz,2H),3.97(s,2H),3.67(dp,J=10.3,7.0Hz,1H),3.20(td,J=7.1,1.0Hz,2H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.1Hz,2H),1.87–1.68(m,4H),1.09–1.02(m,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,155.67,152.90,150.23,146.01,142.56,136.62,129.40,125.49,121.12,118.17,114.98,113.93,112.75,111.08,104.80,90.74,71.28,56.15,51.47,50.56,47.74,30.87,29.91,12.20;ESI-MS m / z:461.2calcd for C 27 H 30 N 6 O 3 [M+H] + 462.2.

[0155] Example 26: 1-(4-(Benzo[d][1,3]dioxazol-5-ylamino)-3-cyano-7-methoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0156]

[0157] In this example, N-(4-amino-2-ethoxyphenyl)acetamide was replaced with 5-amino-2-acetamidobenzyl methyl ether, and p-toluidine was replaced with 5-amino-2,3-dihydrobenzofuran. According to the operation of Example 1, 100 mg of the product was obtained with a yield of 68%; 1 H NMR(400MHz, DMSO-d 6)δ8.76(s,1H),8.60(s,1H),8.37(d,J=9.9Hz,2H),7.43–7.37(m,2H),7.00(dd,J=7.4,1.5Hz,1H),6.78(d,J=7.5Hz,1H),6.06–5.97(m,2H),3.96(s,2H),3.67(dp,J=10.2,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.1Hz,2H),1.82(dq,J=12.4,7.0Hz,2H),1.74(dq,J=12.3,7.0Hz,2H),1.09–1.02(m,3H); 13 CNMR(75MHz,DMSO-d 6 )δ157.63,152.90,150.23,147.91,147.15,146.20,142.58,137.83,129.40,115.86,114.98,113.96,112.75,110.46,104.80,101.45,101.38,90.74,56.15,51.47,50.56,47.74,30.87,12.20;ESI-MS m / z:488.2calcd for C 26 H 28 N 6 O 4 [M+H] + 489.2.

[0158] Example27:Preparation of 1-(3-cyano-7-ethoxy-4-((6-methoxy-5-methylpyridin-3-yl)amino)quinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea

[0159]

[0160] In this example,p-toluidine was replaced with 3-amino-6-methoxy-5-methylpyridine,and 1-ethyl-4-aminopiperidine was replaced with 1-isopropyl-4-piperidinamine.According to the operation of Example 1,80 mg of the product was obtained with a yield of 55%; 1 H NMR(400MHz,DMSO-d 6)δ8.81(s,1H),8.76(s,1H),8.53(s,1H),8.37(s,1H),7.98(d,J=1.4Hz,1H),7.39(s,1H),7.32–7.28(m,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.91(s,2H),3.67(dp,J=10.4,7.0Hz,1H),2.88–2.76(m,3H),2.71(dt,J=12.3,7.1Hz,2H),2.21(s,3H),1.87–1.69(m,4H),1.41(t,J=8.0Hz,3H),1.09(d,J=6.8Hz,5H); 13 C NMR(75MHz,DMSO-d 6 )δ158.34,157.63,151.78,149.11,146.18,142.65,138.72,133.75,129.39,126.58,124.22,114.98,114.13,112.63,105.57,90.74,64.93,54.40,54.22,48.08,47.78,31.03,18.70,14.93,13.83;ESI-MS m / z:517.3calcd for C 26 H 28 N 6 O 4 [M+H] + 518.3.

[0161] Example 28: Preparation of 1-(4-((3-bromo-5-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea

[0162]

[0163] In this example, p-toluidine was replaced with 3-bromo-5-methoxyaniline, and 1-ethyl-4-aminopiperidine was replaced with 1-isopropyl-4-piperidinamine. According to the operation of Example 1, 95 mg of the product was obtained with a yield of 67%; 1 H NMR(400MHz, DMSO-d 6)δ8.71(s,1H),8.60(s,1H),8.45(s,1H),8.39(s,1H),7.39(s,1H),6.96(t,J=1.6Hz,1H),6.78(t,J=1.5Hz,1H),6.43(t,J=1.6Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.82(s,2H),3.66(dp,J=10.3,7.0Hz,1H),2.88–2.76(m,3H),2.71(dt,J=12.4,7.1Hz,2H),1.87–1.69(m,4H),1.40(t,J=8.0Hz,3H),1.09(d,J=6.8Hz,5H); 13 C NMR(75MHz,DMSO-d 6 )δ161.48,157.63,152.31,149.11,145.99,142.48,142.41,129.39,123.34,118.76,114.98,114.95,112.75,112.48,106.66,105.62,90.74,64.57,55.62,54.22,47.95,47.78,31.03,18.70,14.93;ESI-MS m / z:580.2calcd for C 28 H 33 BrN 6 O 3 [M+H] + 581.2.

[0164] Example 29: Preparation of 1-(4-((2-chloro-4-(methylsulfonyl)phenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0165]

[0166] In this example, p-toluidine was replaced with 2-chloro-4-methylsulfonaniline, and according to the operation of Example 1, 95 mg of the product was obtained with a yield of 67%; 1 H NMR(400MHz, DMSO-d 6)δ8.94(s,1H),8.76(s,1H),8.56(s,1H),8.39(s,1H),7.87(d,J=1.5Hz,1H),7.69(dd,J=7.5,1.5Hz,1H),7.44(d,J=7.5Hz,1H),7.40(s,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.65(dp,J=10.2,7.0Hz,1H),3.25(s,2H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.83–1.67(m,4H),1.40(m,3H),1.08–1.01(m,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.63,151.64,149.11,146.39,142.66,141.56,137.95,129.49,128.90,127.21,127.18,122.02,115.00,114.85,112.86,105.55,90.60,64.57,51.36,50.29,47.74,44.63,30.56,14.93,12.20;ESI-MS m / z:570.2calcdfor C 28 H 33 BrN 6 O 3 [M+H] + 571.2.

[0167] Example 30: Preparation of 1-(4-((3-chloro-5-hydroxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea

[0168]

[0169] In this example, p-toluidine was replaced with 3-amino-5-chlorophenol, and according to the operation of Example 1, 90 mg of the product was obtained with a yield of 63%; 1 H NMR(400MHz,DMSO-d 6)δ9.17(s,1H),8.76(s,1H),8.60(s,1H),8.39(s,1H),8.26(s,1H),7.40(s,1H),6.84(t,J=1.6Hz,1H),6.68(t,J=1.6Hz,1H),6.34(t,J=1.6Hz,1H),6.03(d,J=10.3Hz,1H),4.10(q,J=8.0Hz,2H),3.67(dp,J=10.3,7.0Hz,1H),2.86(qt,J=12.5,7.1Hz,4H),2.54(q,J=8.0Hz,2H),1.86(m,4H),1.41(t,J=8.0Hz,3H),1.09–1.02(m,3H); 13 C NMR(75MHz,DMSO-d 6 )δ157.72,157.63,152.31,149.11,146.18,143.59,142.52,135.46,129.39,114.98,114.15,112.96,112.63,111.43,107.13,105.57,90.74,65.05,51.42,50.56,47.74,30.86,14.93,12.20;ESI-MS m / z:508.2calcd forC 26 H 29 ClN 6 O 3 [M+H] + 509.2.

[0170] Example 31: Experiment on the inhibitory activity of HPK1 kinase:

[0171] 1. Experimental method

[0172] The inhibitory activity of the synthesized compound against HPK1 kinase was detected by a luminescence kinase detection method (ADP-Glo TM ) developed by Promega Corporation.

[0173] Specific operation method: First, the compound was serially diluted and transferred to a 384-well white plate. The final starting concentration of the compound was 1000 nM, with a 3-fold serial dilution and 10 concentration points; HPK1 kinase was used with a kinase reaction buffer (HEPES (pH 7.5), 0.01% Tween-20, MgCl 2, diluted to an appropriate concentration with 0.01% BSA and DTT), add enzyme or enzyme reaction buffer to each reaction well, place the reaction plate in a centrifuge, centrifuge at 1000 revolutions per minute, and incubate. Add ATP / substrate mixture to each well, centrifuge at 1000 revolutions per minute, and incubate at room temperature. Add ADP-Glo solution to each well and mix evenly, react at room temperature. Add the detection substrate and incubate at room temperature again. Place it in an enzyme-labeling instrument to read the chemiluminescence signal. Calculate the inhibition rate of the test compound according to the formula (n = 2): Inhibition rate % = (maximum signal value - signal value of each well) / (maximum signal value - minimum signal value) × 100%, IC 50 Obtained by plotting the percentage inhibition rate against the logarithm of the concentration value.

[0174] 2. Experimental results

[0175] The pharmacological experimental results of some preferred compounds are as follows:

[0176] Table 1. Experimental data of the compounds on HPK1 kinase activity

[0177]

[0178] Note: a The compounds were tested for HPK1 kinase activity at a concentration of 1.0 μM.

[0179] As can be seen from Table 1, the IC of the compounds designed in the present invention for inhibiting HPK1 50 value reaches the nanomolar concentration level, and the kinase activity of HPK1 at the micromolar concentration level is lower than 40%, showing excellent inhibitory activity.

[0180] Example 32: Water solubility experiment of some compounds

[0181] 1. Experimental method

[0182] (1) Experimental instruments and conditions

[0183] HPLC model: Agilent 1100; chromatographic column: Lichrospher C18 (5 μm, 4.6 × 250 mm); column temperature: 37 °C; flow rate: 1.0 ml / min; injection volume: 10 μL; detection wavelength: 210 nm.

[0184] (2) Experimental steps

[0185] Accurately weigh each sample, prepare standard solutions with concentrations of 0.01, 0.05, 0.1, 0.2, 0.4, and 1 mg / mL using methanol, inject the samples into HPLC to obtain the peak areas of the main peaks, and calculate the standard curve. Then, take an appropriate excess of the compound to be tested (>10 mg), dissolve it in PBS respectively, and incubate it in a constant temperature shaker at 37 °C for 24 hours. Centrifuge and take 10 μL of the supernatant and inject it into HPLC to measure the peak areas of each compound, and then calculate the corresponding solubility through the standard curve.

[0186] 2. Experimental results

[0187] Table 2. Water solubility results of some compounds of the present invention

[0188] Example μg / ml in water-soluble (PBS pH 7.4) 11 85 21 79 27 102 30 105

[0189] As can be seen from Table 2, the compounds designed in the present invention have good water solubility, which is beneficial for drug formation.

[0190] Example 33: Liver microsome stability experiment

[0191] 1. Experimental method

[0192] Pre-incubate MgCl 2 / NADPH / different species of liver microsomes (human, rat, and mouse liver microsomes) / PBS in a 37 °C water bath for 5 min, then add the corresponding compound solution to start the reaction, and the incubation time is 0 min, 5 min, 15 min, 30 min, 60 min, and 120 min. The content of organic solvent in the whole system is <1%. After the reaction is completed, add 400 μL of ice methanol to terminate the reaction and immediately take it out. Detect the samples according to the corresponding LC-MS / MS detection method, and process the detection results using Graphpad prism 8.3.0 to calculate the phase I metabolic stability parameter t of the compound in different species of liver microsomes 1 / 2 (min).

[0193] 2. Experimental results

[0194] Table 3. Liver microsome stability of some compounds

[0195]

[0196]

[0197] As can be seen from Table 3, the compounds designed in the present invention have good metabolic stability in vivo, which is beneficial for drug formation.

Claims

1. A 3-cyano-substituted quinoline compound, characterized in that, it has the structure of formula I and also includes its pharmaceutically acceptable salts, wherein: R 1 selected from C 1 -C 6 alkyl; R 2 selected from pyridine, 5-6 membered aromatic ring, 6 membered fused 6 membered aromatic ring, quinoline, isoquinoline, indole, benzofuran; R 2 substituted by 1 to 4 identical or different Rs; R is selected from a hydrogen atom, a halogen, a hydroxyl group, a nitro group, C 1 -C 6 -sulfonyl, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy; the C 1 -C 6 -alkyl and C 1 -C 6 -alkoxy are substituted by one or two or three substituents selected from halogens; R 3 selected from hydrogen, deuterium, C 1 -C 6 -alkyl, saturated C3-C7 cycloalkyl, or R 3 is absent; X is selected from an oxygen atom, NH, and a sulfur atom; Y is selected from CH, an oxygen atom, a nitrogen atom, and a sulfur atom.

2. A 3-cyano-substituted quinoline compound, characterized in that, in the said structure: R 1 selected from C 1 -C 6 alkyl; R 2 Selected from 5- or 6-membered aromatic ring pyridine, 6-membered ring-fused 6-membered aromatic ring, quinoline, isoquinoline, indole, benzofuran; R 2 substituted by 1-4 identical or different Rs; R is selected from a hydrogen atom, a halogen, a nitro group, C 1 -C 6 alkyl, C 1 -C 6 alkoxy; R 3 selected from hydrogen, deuterium, C 1 -C 6 -alkyl, saturated C3-C7 cycloalkyl, or R 3 is absent; X is selected from an oxygen atom, NH, and a sulfur atom; Y is selected from CH, an oxygen atom, a nitrogen atom, and a sulfur atom.

3. The 3-cyano-substituted quinoline compound according to claim 1 or 2, characterized in that, in the said structure: R 3 selected from hydrogen, C 1 -C 6 -alkyl, saturated three-membered cyclic alkyl, or R 3 is absent; X is selected from an oxygen atom and a sulfur atom; Y is selected from CH, an oxygen atom, and a nitrogen atom.

4. The 3-cyano-substituted quinoline compound according to claim 2, characterized in that, in the said structure: R 1 selected from C 1 -C 6 alkyl; R 2 selected from a 6-membered aromatic ring, pyridine, quinoline, isoquinoline, indole, benzofuran; R 2 substituted by 1-4 identical or different Rs; R is selected from halogen, nitro, C 1 -C 6 alkyl, C 1 -C 6 alkoxy group.

5. The 3-cyano-substituted quinoline compound according to claim 4, characterized in that, in the said structure: R 1 selected from C 1 -C 4 alkyl; R 2 selected from a 6-membered aromatic ring, pyridine, quinoline, isoquinoline, indole; R 2 substituted by 1 to 4 identical or different Rs; R is selected from a hydrogen atom, a halogen, a nitro group, C 1 -C 6 alkyl, C 1 -C 6 alkoxy; R 3 selected from hydrogen, C 1 -C 6 -alkyl, saturated three-membered cyclic alkyl, or R 3 is absent; X is selected from an oxygen atom and Y is selected from a nitrogen atom and an oxygen atom.

6. A 3-cyano-substituted quinoline compound, characterized in that, in the said structure: R 1 selected from methyl, ethyl; R 2 selected from R 3 selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, or R 3 absent; X is selected from an oxygen atom, NH, and a sulfur atom; Y is selected from CH, an oxygen atom, a nitrogen atom, and a sulfur atom.

7. A 3-cyano-substituted quinoline compound, characterized in that, it is selected from any one of the following compounds: 1-(3-cyano-7-ethoxy-4-(p-benzylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (1), 1-(3-cyano-7-ethoxy-4-((4-nitrophenyl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (2), 1-(4-((4-chloro-2-fluorophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (3), 1-(3-cyano-4-((2,4-difluoro-5-methoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (4), 1-(3-cyano-7-ethoxy-4-((2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (5), 1-(3-cyano-7-ethoxy-4-(phenylamino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (6), 1-(3-cyano-7-ethoxy-4-((1-methyl-1H-indol-6-yl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (7), 1-(3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (8), 1-(3-cyano-4-((2,4-dichlorophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (9), 1-(4-((1H-indazol-5-yl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (10), 1-(4-((2-chloro-5-nitrophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (11) 1-(4-((2-chloro-4-fluorophenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (12) 1-(4-((2-chloro-5-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea (13) 1-(3-cyano-4-((2,4-dichlorophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(1-cyclopropylpiperidin-4-yl)urea (14) 1-(4-((4-chloro-3-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-methylpiperidin-4-yl)urea (15) 1-(4-((4-chloro-3-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea (16) 1-(4-((4-bromo-2-methylphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea (17) 1-(3-cyano-4-((2,4-diiodophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea (18) 1-(3-cyano-4-((4,5-dimethyl-2-nitrophenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea (19) 1-(3-cyano-4-((3,4-dimethoxyphenyl)amino)-7-ethoxyquinolin-6-yl)-3-(tetrahydro-2H-thiopyran-4-yl)urea (20) 1-(3-cyano-7-methoxy-4-((4-methoxy-3-(trifluoromethyl)phenyl)amino)quinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (21) 1-(4-((6-bromonaphthalen-2-yl)amino)-3-cyano-7-methoxyquinolin-6-yl)-3-(1-methylpiperidin-4-yl)urea (22) 1-(4-((2-chloro-5-methoxyphenyl)amino)-3-cyano-7-methoxyquinolin-6-yl)-3-cyclohexylurea (23), 1-(3-cyano-7-methoxy-4-((4-methoxy-3-methylphenyl)amino)quinolin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)urea (24) 1-(3-cyano-4-((2,3-dihydrobenzofuran-5-yl)amino)-7-methoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (25) 1-(4-(benzo[d][1,3]dioxol-5-ylamino)-3-cyano-7-methoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (26) 1-(3-cyano-7-ethoxy-4-((6-methoxy-5-methylpyridin-3-yl)amino)quinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea (27), 1-(4-((3-bromo-5-methoxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-isopropylpiperidin-4-yl)urea (28), 1-(4-((2-chloro-4-(methylsulfonyl)phenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (29), 1-(4-((3-chloro-5-hydroxyphenyl)amino)-3-cyano-7-ethoxyquinolin-6-yl)-3-(1-ethylpiperidin-4-yl)urea (30).

8. The 3-cyano-substituted quinoline compound according to claim 1, wherein, the pharmaceutically acceptable salt is a salt formed by the compound and an acid or a base, the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, fumaric acid, succinic acid, salicylic acid, phenylacetic acid or mandelic acid, and the base is a base containing sodium, potassium, calcium, ammonium cations, triethylamine, diethylamine or choline.

9. A pharmaceutical composition, wherein, it comprises the 3-cyano-substituted quinoline compound according to claim 1 and a pharmaceutically acceptable carrier.

10. Use of the 3-cyano-substituted quinoline compound according to claim 1 or the pharmaceutical composition according to claim 9 in the preparation of an HPK1 inhibitor drug.

11. The use according to claim 10, wherein, the drug is a drug for preventing and / or treating cancer, autoimmune diseases or immune-mediated diseases.

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