N-substituted aromatic ring-azabicyclo amine derivative and medical application

Abstract

The invention discloses a compound as shown in a formula (I) and medical application, including optical isomers and pharmaceutically acceptable salts thereof. The compound disclosed by the invention has FLT3 and IRAK4 kinase inhibition activity, has proliferation inhibition activity on FLT3-ITD positive leukemia cell strains MV4-11 and MOLM13, also has proliferation inhibition activity on an IRAK4 positive leukemia cell strain TF-1, can overcome adaptive drug resistance of a clinical FLT3 inhibitor, and can be applied to preparation of antitumor drugs.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to an N-substituted aromatic ring-azabicyclic amine derivative and its pharmaceutical uses, optical isomers, salts and preparation methods, as well as its use as an inhibitor of Fms-like tyrosine kinase 3 (FLT3), interleukin-1 receptor-associated kinase 4 (IRAK4) or dual-targeting FLT3 / IRAK4 in antitumor drugs. Background Technology

[0002] FLT3 (FMS-like tyrosine kinase 3) is a transmembrane receptor tyrosine kinase protein typically expressed by hematopoietic stem cells. It plays a crucial role in cell development, promoting cell survival, growth, and differentiation through various signaling pathways. FLT3 gene mutations can drive oncogenic signaling and are one of the most common genetic abnormalities in AML, affecting approximately 30% of AML patients. Studies have found that activated FLT3 promotes AML cell proliferation and differentiation through the PI3K, RAS, and STAT5 signaling pathways. There are two main types of FLT3 mutations: internal tandem repeat mutations in the juxtamembrane domain (FLT3-ITD) and point mutations or deletions in the tyrosine kinase domain (FLT3-TKD). Both types of mutated FLT3 molecules can be activated by ligand-independent dimerization and transphosphorylation. FLT3-ITD is the most common FLT3 mutation, affecting approximately 25% of AML patients. Compared to AML patients without the FLT3-ITD mutation, FLT3-ITD AML patients have a worse overall prognosis, including an increased relapse rate, an increased risk of death after relapse, and a higher likelihood of relapse after hematopoietic stem cell transplantation. Given the important role of FLT3 mutations in the development and progression of AML, FLT3-targeted therapy has become a hot research topic.

[0003] Currently, several FLT3 inhibitors (FLT3i) are on the market, among which Midostaurin, Gilteritinib, and Quizaritinib have been approved for the treatment of AML patients with FLT3 mutations. Although these drugs have demonstrated excellent initial clinical responses in AML patients as monotherapy or in combination, short response times and high relapse rates due to mutations in the FLT3-kinase domain (acquired resistance) and activation of bypass pathways (adaptive resistance) remain major challenges in the treatment of AML patients. Developing treatments that can overcome both acquired and adaptive resistance is essential.

[0004] Tumor immunotherapy is currently a hot topic in medical research. It utilizes the regulatory mechanisms of the immune system to recognize and kill tumor cells, and is considered an important direction for future cancer treatment. Interleukin-1 receptor-associated kinase 4 (IRAK4) is a key signaling node transducing interleukin-1 receptor and T-lymphocyte-like receptor family signals. Studies have shown that IRAK4-L is highly expressed in 50% of MDS and AML patients and is highly correlated with poor prognosis. After FLT3i treatment, TLR expression increases on FLT3-ITD AML cells, and the innate immune pathway (IRAK1 / 4) is compensatorily activated, which can further restore Ras / MAPK and NF-κB, ultimately leading to cell survival and adaptive resistance. Therefore, FLT3 / IRAK4 dual-target inhibitors can overcome adaptive resistance in AML treatment by targeting the immune stress response pathway. Developing novel small molecule inhibitors with excellent FLT3 / IRAK4 kinase inhibitory activity is of great significance for the treatment of hematologic malignancies. Summary of the Invention

[0005] The purpose of this invention is to provide a class of N-substituted aromatic ring-azabicyclic amine derivatives with FLT3 and IRAK4 inhibitory activities and antitumor effects, as well as their pharmaceutical uses. These compounds exhibit excellent biological activity while enhancing the molecular diversity and novelty of the compounds. Another purpose of this invention is to provide a method for preparing such N-substituted aromatic ring-azabicyclic amine derivatives and their pharmaceutical uses, as well as their applications.

[0006] The technical solution of the present invention is as follows:

[0007] Compounds of general formula (I) or their pharmaceutically acceptable salts:

[0008]

[0009] Or its optical isomer or its pharmaceutically acceptable salt.

[0010] Wherein, A is selected from a benzene ring or a 5-6 membered heteroaromatic ring, wherein the heteroatom in the heteroaromatic ring is 1-4 N atoms, and the benzene ring or heteroaromatic ring may be further replaced by 1-3 R4 atoms;

[0011] X is selected from CH or N;

[0012] Y is selected from N and CR. X ;R X Selected from H, deuterium, halogens, C 1-3 alkyl, C 1-3 Alkyloxy groups, halogenated C 1-3 Alkyl, CN, NO2;

[0013] R 1 Selected from H, deuterium, halogens, C1-3 Alkyl, halogen-substituted C 1-3 Alkyl, C 1-3 Alkyloxy group, -NR a R b -C(=O)-NR a R b -S(=O)2-NR a R b ;

[0014] R 2 R 3 Each is independently selected from H, deuterium, halogen, and C. 1-3 Alkyl, halogen-substituted C 1-3 Alkyl, C 1-3 Alkyloxy group, CN, NO2, -C(=O)-NR aa R bb -S(=O)2-NR aa R bb -L 1 -NR aa R bb -L 1 -R 5 L 1 It is a chemical bond, -O(CH2) o -、-NR 7 (CH2) p -;

[0015] R a R b R aa R bb Each is independently selected from H, deuterium, OH, and C. 1-3 alkyl, C 3-5 cycloalkyl, -(C=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups);

[0016] o and p are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8;

[0017] R 4 Selected from H, C 1-5 alkyl, C 3-8 Cycloalkyl groups, 4- to 8-membered heterocyclic alkyl groups, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups, phenyl groups, and 5-8 heteroaryl groups, wherein the alkyl, cycloalkyl, phenyl, and heteroaryl groups can be independently separated by 0, 1, or 2 R groups. 8 Replaced by multiple R 8When replacing, multiple R 8 They can be independent and may be the same or different;

[0018] R 5 Selected from H, OH, CN, halogens, C 1-3 alkyl, C 1-3 Halogenated alkyl, C 4-8 Cycloalkyl groups, 4- to 8-membered heterocycloalkyl groups, wherein the alkyl, cycloalkyl, or heterocycloalkyl group may be further substituted with one or two R groups. 6 replace;

[0019] R 6 Selected from H, NH2, -NH-C 1-3 Alkyl, -N-(C 1-3 Alkyl)2, C 1-5 alkyl, C 3-8 Cycloalkyl groups, 4- to 8-membered heterocyclic alkyl groups, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 The alkyl group, wherein the cycloalkyl group may be further substituted with 0, 1 or 2 hydroxyl groups;

[0020] R 7 For H, C 1-3 Alkyl groups;

[0021] R 8 For H, deuterium, halogen, OH, NH2, CF3, OCH3, CN, NO2, C 1-5 Alkyl group, -C(=O)NH-L 2 -R 9 The alkyl group may be further substituted with 0, 1, or 2 hydroxyl groups or amino groups;

[0022] L 2 For chemical bonds, C 1-8 alkylene s and t are independently selected from 1, 2, and 3, respectively;

[0023] R 9 The components are H, OH, and NH2.

[0024] As a preferred option

[0025] A is selected from:

[0026]

[0027] in Represents the substituent junction; the R 4 Defined as general formula (I).

[0028] R X R1 R 2 R 3 C 1-3 The alkyloxy group is preferably OCH3; the R a R b R aa R bb C 3-5 The cycloalkyl group is preferably cyclopropyl or cyclobutyl.

[0029] Preferably, it has the structure shown in general formula II:

[0030]

[0031] The R 1 R 2 R 3 R 4 The definition is the same as that shown in any of the above technical solutions.

[0032] Furthermore, R 1 H, -NR a R b ;

[0033] R 2 For -NH(CH2) p -NR aa R bb -NR 7 (CH2) p -R 5 、;

[0034] R a R b Each is independently selected from H, deuterium, methyl, and ethyl;

[0035] p is selected from 0, 1, 2, 3, 4, 5;

[0036] R 3 It consists of H, deuterium, and halogens;

[0037] R 4 For H, C 1-5 alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups, -S(=O)2-(C 1-3 Alkyl groups, phenyl groups, and 5-8 membered aromatic heterocycles, wherein the alkyl, cycloalkyl, phenyl, and aromatic heterocycles can be independently separated by 0, 1, or 2 R groups. 8 replace;

[0038] R 5 It is a 4-8 heterocyclic alkyl group; the heterocyclic alkyl group may be further divided by 0, 1 or 2 R groups. 6 replace;

[0039] R 6 For H, NH2, -NH-C 1-3 Alkyl, -N-(C 1-3 alkyl)2, methanesulfonyl, C 1-4 Alkyl groups, 4- to 6-membered heterocyclic alkyl groups, wherein the alkyl group may be further substituted with 0 or 1 hydroxyl group;

[0040] R 7 For H and CH3;

[0041] R 8 For H, deuterium, halogen, OH, CF3, OCH3, C 1-5 The alkyl group may be further substituted with 0 or 1 hydroxyl group.

[0042] As a preferred embodiment, the compound has the following structure:

[0043] R 2 For -NH(CH2) p -NR aa R bb , R aa R bb Each is independently selected from H, deuterium, and CH3; m, n, and p are independently selected from 1, 2, 3, 4, and 5; R 6 H, methanesulfonyl, C 1-4 The alkyl group or 3- to 6-membered heterocyclic alkyl group may be further substituted with hydroxyl groups.

[0044] In this invention, "alkyl" and the alkyl portion of other groups (e.g., alkoxy) can be straight-chain or branched, generally referring to C 1-5 Alkyl groups (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc.). The C of this invention... 1-3 Alkyl groups generally include, for example, methyl, ethyl, n-propyl, isopropyl, etc.

[0045] In this invention, "cycloalkyl" refers to a monocyclic or polycyclic hydrocarbon ring group, generally referring to a 3-10 member cycloalkyl group, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclohexadienyl, cyclohepttrienyl, borneol, norpinyl, norcaryl, adamantyl, benzoyl, decahydronaphthyl, norbornyl, spiro[4.5]decyl, etc. The "C" mentioned in this invention... 3-8 "Cycloalkyl" generally includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. Cycloalkyl groups can be unsubstituted or substituted with one or more suitable substituents.

[0046] In this invention, "halogen" includes fluorine, chlorine, bromine, or iodine.

[0047] In this invention, "heterocyclic alkyl" and "aromatic heterocyclic" refer to a ring system in which at least one carbon atom member is replaced by at least one heteroatom such as nitrogen, sulfur, or oxygen.

[0048] In this invention, "heterocyclic alkyl" refers to a non-aromatic monocyclic or polycyclic compound containing carbon and hydrogen atoms, as well as at least one heteroatom (preferably 1-4 heteroatoms from nitrogen, oxygen, sulfur, sulfone, or sulfoxide). Heterocyclic alkyl groups may have one or more carbon-carbon double bonds or carbon-heteroatom double bonds in the cyclic group, as long as the cyclic group does not become aromatic by its presence. The total number of carbon atoms and heteroatoms on the ring is generally 3-12. For example, the 4-8 membered heterocyclic alkyl groups generally include azirhexacyclobutane, piperidinyl, piperazine, pyrrolidinyl, etc.

[0049] In one embodiment of the present invention, A may specifically be selected from the following structure:

[0050]

[0051] In one embodiment of the present invention, X may specifically be CH or N.

[0052] In one embodiment of the present invention, Y can specifically be CH or N.

[0053] In one embodiment of the present invention, R 1 Specifically, H or -NHCH2CH3 can be selected.

[0054] In one embodiment of the present invention, R 2 Specifically, -NH(CH2) can be selected. p -NR aa R bb -NR 7 (CH2) p -R 5 Furthermore, R 2 Selected from:

[0055] In one embodiment of the present invention, R a R b Specifically, they can be selected independently from H and CH3.

[0056] In one embodiment of the present invention, p is selected from 0, 1, 2, 3, 4, and 5.

[0057] In one embodiment of the present invention, R 3 Specifically, H and F are optional.

[0058] In one embodiment of the present invention, R 4 Specifically, the following can be selected: H, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, 3-oxetanebutyl, hydroxytert-butyl, methanesulfonyl, 2-methylprop-2-ol, pyridine ring, N-methylpyrazole ring, benzene ring, substituted benzene ring (such as phenyl with one or two halogens substituted, methylphenyl, methoxyphenyl, trifluoromethylphenyl, hydroxyisopropyl-substituted phenyl, carbamoylphenyl, hydroxy and methyl-substituted butylcarbamoylphenyl), methylimidazolyl.

[0059] In one embodiment of the present invention, R 5 Morpholine is a specific option.

[0060] In one embodiment of the present invention, R 6 Specifically, the following can be selected: H, CH3, 3-oxetanebutyl, methanesulfonyl, or 2-methylprop-2-ol.

[0061] In one embodiment of the present invention, R 7 Specifically, H or methyl can be selected.

[0062] In one embodiment of the present invention, the compound of general formula I may be selected from the following specific compounds:

[0063]

[0064]

[0065]

[0066] And optical isomers or pharmaceutically acceptable salts.

[0067] The numbers below the above 55 compounds are their corresponding codes. For ease of description and brevity, these codes will be used directly in the following content of this specification.

[0068] This invention also provides a method for preparing the above-mentioned N-substituted aromatic ring-azabicyclic amine derivatives, their pharmaceutical uses, optical isomers, or pharmaceutically acceptable salts thereof, through the following steps:

[0069] The reaction route of the preparation method is shown in the following equation:

[0070]

[0071] Where Z = Br, I; R 1 R 2 R 3 The definitions of X, Y, and A are as described in claim 1; R 2’ For R2 Or R 2 The Boc protected precursor; the preparation method includes the following steps:

[0072] (1) Compound 1 and an amine compound (such as 1-N-Boc-cis-1,4-cyclohexanediamine, 1-N-Boc-trans-1,4-cyclohexanediamine, N-Boc-1,3-propanediamine, N-Boc-1,4-butanediamine) are dissolved in a reaction solvent (such as dioxane). Under inert gas protection, Pd2(dba)3, Xant-phos and Cs2CO3 are added and reacted at 100℃~120℃ (for example, at 110℃). After the reaction is completed, compound 2 is obtained.

[0073] In step (1), after the reaction was completed, the solvent was removed by vacuum distillation using TLC, and compound 2 was purified by silica gel column chromatography.

[0074] (2) Compound 2 was dissolved in methanol, 10% Pd / C was added at room temperature, and the reaction was carried out at 50°C under the action of hydrogen. The reaction was completed, and compound 3 was obtained after post-treatment.

[0075] In step (2), after the reaction was detected by TLC, the reaction mixture was filtered through diatomaceous earth, the solvent was removed by vacuum distillation, and the mixture was subjected to silica gel column chromatography to finally obtain compound 3.

[0076] (3) Dissolve compounds 3 and 4 in a reaction solvent (e.g., dioxane), add Pd2(dba)3, X-phos and K2CO3 under inert gas protection, and react at 100℃~120℃ (e.g., at 110℃). After the reaction is completed, process to obtain compound I or the Boc protected precursor of compound I. The Boc protected precursor is further de-Boc grouped with trifluoroacetic acid to obtain compound I.

[0077] In step (2), the solvent is removed by vacuum distillation after the reaction is completed using TLC detection, and the compound I or its Boc protected precursor is purified by silica gel column chromatography.

[0078] Specifically, the following methods can be used:

[0079] Method 1: The synthetic route for N-substituted aromatic ring-azabicyclic amine derivatives and their pharmaceutical applications is as follows:

[0080] Depending on the A ring, the synthetic route varies slightly, as shown in (1) and (2):

[0081] (1) When A is When the six-membered ring is used, its synthetic route is as follows:

[0082]

[0083] The preparation method (1) of the above-mentioned N-substituted aromatic ring-azabicyclic amine derivatives and pharmaceutical applications includes the following steps:

[0084] 1) Raw material 1 and the corresponding amine under N2 protection, base, palladium and corresponding ligand catalysis conditions, overnight at 110°C, underwent Buchwald-Hartwig coupling reaction to obtain intermediate 1. The solvent used for Buchwald-Hartwig coupling was anhydrous dioxane, the base was cesium carbonate, the catalyst was Pd2(dba)3, and the ligand was Xantphos.

[0085] 2) Intermediate 1 was reacted with 10% Pd / C and hydrogen for 4 h to obtain intermediate 2. The reaction solvent was methanol and the reaction temperature was 50℃.

[0086] 3) Intermediate 2 and raw material 2 undergo a Buchwald-Hartwig coupling reaction overnight at 100°C under N2 protection, base, palladium and corresponding ligand catalysis. Subsequently, the Boc protecting group is removed to obtain compound I-1. The solvent used for Buchwald-Hartwig coupling is anhydrous dioxane, the base is potassium carbonate, the catalyst is Pd2(dba)3, and the ligand is Xphos.

[0087] (2) When A is When the five-element ring is in use, For example, its synthetic route is as follows:

[0088]

[0089] The preparation method (2) of the above-mentioned N-substituted aromatic ring-azabicyclic amine derivatives and pharmaceutical applications includes the following steps:

[0090] 1) Raw material 1 and the corresponding amine under N2 protection, base, palladium and corresponding ligand catalysis conditions, overnight at 110°C, underwent Buchwald-Hartwig coupling reaction to obtain intermediate 1. The solvent used for Buchwald-Hartwig coupling was anhydrous dioxane, the base was cesium carbonate, the catalyst was Pd2(dba)3, and the ligand was Xantphos.

[0091] 2) Intermediate 1 was reacted with 10% Pd / C and hydrogen for 4 h to obtain intermediate 2. The reaction solvent was methanol and the reaction temperature was 50℃.

[0092] 3) Raw material 3 undergoes a substitution or oxidative cross-coupling reaction with the corresponding iodide or arylboronic acid under alkaline conditions to obtain intermediate 3. When substituted with an iodide, the base is sodium hydrogen, the reaction solvent is THF, and the reaction condition is room temperature for 5 hours; when coupled with an arylboronic acid, the base is triethylamine, the catalyst is copper acetate, the reaction solvent is DCM, and the reaction condition is overnight at room temperature in contact with oxygen.

[0093] 3) Intermediates 2 and 3 under N2 protection, base, palladium and corresponding ligand catalysis, underwent Buchwald-Hartwig coupling reaction overnight at 100°C, followed by removal of the Boc protecting group to obtain compound I-2. The solvent used for Buchwald-Hartwig coupling was anhydrous dioxane, the base was potassium carbonate, the catalyst was Pd2(dba)3, and the ligand was Xphos.

[0094] 4) The aminopiperidine-containing compounds in Formula I compound-2 (defined as Formula I compound-3) undergo substitution or reductive amination reactions with the corresponding halogens or ketones under alkaline conditions to yield Formula I compound-4. When substituted with a halogen, the base is triethylamine, the reaction solvent is DCM, and the reaction condition is room temperature for 4 hours. When reductively amination occurs with a ketone, the catalyst is acetic acid, the reducing agent is sodium cyanoborohydride, the reaction solvent is DMF, and the reaction condition is room temperature overnight.

[0095] Method 2:

[0096] The compound of the present invention was dissolved in isopropanol, and a solution containing 1 to 1.5 times (e.g., 1.2 times equivalent) of an organic or inorganic acid was slowly added dropwise at room temperature. After the addition was complete, the reaction solution was stirred overnight at 40°C. The reaction solution was cooled to room temperature, filtered, and the solid was washed with diethyl ether and dried to obtain the corresponding salt.

[0097] In the above preparation process, Z is a bromine or iodine atom, and R... 1 R 2 R 3 R 4 R 6 The definitions of X, Y, and R are as shown above; 2’ For R 2 Or R 2 The group before the removal of the Boc protecting group;

[0098] Those skilled in the art can prepare the compounds of the present invention using various starting compounds conventionally obtained in the art as raw materials, according to actual preparation needs.

[0099] In one embodiment of the present invention, pharmaceutically acceptable salts are organic and inorganic salts. Organic salts include acetates, formates, propionates, pyruvates, glycolates, oxalates, oxalates, malates, succinates, glutarates, mandelates, citrates, trifluoroacetates, fumarates, oxalates, malates, L-malates, D-malates, lactates, camphor sulfonates, p-toluenesulfonates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, benzoates, tartrates, L-tartrates, D-tartrates, oxalates, succinates, maleates, ascorbic acid salts, and amino acid salts (such as aspartate). Inorganic salts include hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, hydroiodates, and perchlorates.

[0100] This invention provides the use of the above-mentioned compounds as Fms-like tyrosine kinase 3 (FLT3), interleukin-1 receptor-associated kinase 4 (IRAK4), or dual-targeting FLT3 / IRAK4 inhibitors in antitumor drugs.

[0101] In one embodiment of the invention, the invention provides a method for treating a protein kinase-mediated disease or condition, comprising administering to an individual in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition, or an agent thereof. The protein kinase includes, but is not limited to, FLT3 (including mutant forms FLT3-ITD, FLT3-D835Y / V / F / I / N, FLT3-F691L / I, FLT3-ITD-D835Y / V / F, FLT3-ITD-F691L / I, FLT3-N676D, FLT3-Y842H / R, FLT3-G697R, FLT3-D815H / N, FLT3-K633Q) and IRAK4.

[0102] In one embodiment of the present invention, diseases associated with the aforementioned kinase include tumors and autoimmune diseases. The tumors mentioned include bladder cancer, breast cancer, colon cancer, kidney cancer, epidermal cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, or skin cancer; lymphatic spectrum hematopoietic tumors such as acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, pilocellular lymphoma, Burkett's lymphoma, bone marrow spectrum hematopoietic tumors, acute and chronic myeloid leukemia, acute and chronic myeloid leukemia, spinal dysplasia syndrome, promyeloid leukemia, thyroid follicular carcinoma, stromal tumors, fibrosarcoma, rhabdomyosarcoma, central or peripheral nervous system tumors, astrocytoma, neuroblastoma, glioma, schwannoma, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, xanthokeratoma, thyroid follicular carcinoma, and Kaposi's sarcoma.

[0103] In one embodiment of the present invention, the immune disease is selected from arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Auder's thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuritis, acute infectious encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis, graft-versus-host disease, transplantation, transfusion allergy, allergic reaction, type I hypersensitivity reaction, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

[0104] In one embodiment of the invention, the active compound of the invention or a pharmaceutically acceptable salt thereof may be used alone as the sole antitumor drug, or may be used in combination with other therapeutic agents. These other therapeutic agents are selected from IDH1 inhibitors, IDH2 inhibitors, Bcl-2 inhibitors, hypomethylating agents, and antimetabolites.

[0105] The compounds of this invention have FLT3 and IRAK4 inhibitory activities, and exhibit proliferative inhibitory activity against FLT3-ITD positive leukemia cell lines MV4-11 and MOLM13, as well as proliferative inhibitory activity against IRAK4 positive leukemia cell line TF-1. They can overcome adaptive resistance to FLT3 inhibitors in clinical practice and can be used in the preparation of antitumor drugs. Detailed Implementation

[0106] The present invention will be further described in conjunction with embodiments.

[0107] The preparation of compounds will be further illustrated in the following examples. These examples are for illustrative purposes only and do not limit the invention in any way.

[0108] Preparation Example 1: (R)-N 1 -(7-Ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 1)

[0109]

[0110] Step 1: Synthesis of ((R)-3-((3-fluoro-5-nitrophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 1-1)

[0111] Under nitrogen protection, a mixture of 1-bromo-3-fluoro-5-nitrobenzene (300 mg, 1.37 mmol), (R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (357 mg, 1.78 mmol), tris(dibenzylindeneacetone)palladium (62 mg, 0.068 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (81 mg, 0.14 mmol), and cesium carbonate (1114 mg, 3.42 mmol) was added to anhydrous dioxane (8 mL). The mixture was reacted overnight at 110 °C. After filtration, the solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 8:1 as the eluent to give a yellow oil 1-1 (340 mg, 73%). 1 HNMR(500MHz,CDCl3)δ7.23(d,J=2.0Hz,1H),7.19(dt,J=8.5,2.0Hz,1H),6.59(dt,J=10.5,2.0Hz,1H),4.34-4.23(s,1H),3.933.86(m,1H),3.64 (m,1H),3.43(m,1H),3.20(m,1H),3.07(m,1H),2.00(m,1H),1.80-1.70( m,1H),1.60(dd,J=12.5,6.0Hz,2H),1.46(s,9H); ESI-MS:m / z=340[M+H] + .

[0112] Step 2: Synthesis of (R)-3-((3-amino-5-fluorophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 1-2)

[0113] At room temperature, intermediate 1-1 (340 mg, 1.00 mmol) was dissolved in anhydrous methanol (6 ml), and 10% Pd / C (35 mg) was added. The mixture was reacted at room temperature for 4 h under the action of hydrogen. After the reaction was complete, the reaction solution was filtered with diatomaceous earth, and the solvent was recovered under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography with PE:EA = 3:1 as the eluent to obtain a pale yellow oily substance 1-2 (281 mg, 91%). 1 HNMR(500MHz,CDCl3)δ5.80(m,1H),5.78(m,1H),5.72(s,1H),3.96(s,1H),3 .72–3.56(m,4H),3.29(m,1H),3.13–3.02(m,1H),2.89(m,1H),1.95(m,1H),

[0114] 1.75–1.68(m,1H),1.46(s,9H); ESI-MS: m / z=310[M+H] + .

[0115] Step 3: Synthesis of 2-chloro-7-ethyl-7H-pyrrolo[2,3-d]pyrimidine (intermediates 1-3)

[0116] Dissolve 100 mg (0.65 mmol) of 2-chloro-7H-pyrrolo[2,3-d]pyrimidine in 3 mL of THF. Slowly add 20 mg (0.85 mmol) of NaH under ice bath conditions. Stir at room temperature for 0.5 h, then add 406 mg (2.6 mmol) of iodoethane. Continue the reaction at room temperature for 5 h. After the reaction is complete, the solvent is recovered under reduced pressure to obtain the residue. Purify by silica gel column chromatography with PE:EA = 6:1 as the eluent to give a white solid 1-3 (89 mg, 75%). 1 H NMR (500MHz, CDCl3) δ8.78 (s, 1H), 7.24 (d, J = 3.5Hz, 1H), 6.56 (d, J = 3.5Hz, 1H), 4.30 (q, J = 7.5Hz, 2H), 1.48 (t, J = 7.5Hz, 3H); ESI-MS: m / z =

[0117] 182 [M+H] + .

[0118] Step 4: (R)-N 1 -(7-Ethyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 1)

[0119] Under nitrogen protection, a mixture of intermediates 1-2 (111 mg, 0.36 mmol), intermediates 1-3 (60 mg, 0.33 mmol), tris(dibenzylindeneacetone)palladium (30 mg, 0.033 mmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (31 mg, 0.066 mmol), and potassium carbonate (137 mg, 0.99 mmol) was added to anhydrous dioxane (3 mL). The mixture was reacted overnight at 100 °C. After filtration, the solvent was recovered under reduced pressure to obtain the residue, which was then purified by silica gel column chromatography.

[0120] Using PE:EA = 2:1 as the eluent, 105 mg of a pale yellow solid was obtained. This solid was dissolved in DCM (2 ml), and 0.5 ml of trifluoroacetic acid was added dropwise under ice bath conditions. After reacting the mixture at room temperature for 5 h, the solvent was recovered by vacuum distillation to obtain a yellow crude product. This product was then purified by silica gel column chromatography using DCM:NH3 / MeOH (30:1-20:1) as the eluent to obtain a white solid (49 mg, 42%). 1H NMR (500MHz, DMSO-d6) δ9.29(s,1H),8.67(s,1H),7.30(d,J=3.5Hz,1H),7.04(d,J=2.0Hz,1H),6.93(dt,J=12.0,2 .0Hz,1H),6.43(d,J=3.5Hz,1H),5.95(dt,J=12.0,2.0Hz,1H),5.58(d,J=8.0Hz,1H),4.19(q,J=7.0Hz,2H),3.22( m,1H),3.09(dd,J=12.0,4.0Hz,1H),2.79(dd,J=12.0,4.0Hz,1H),2.48–2.42(m,1H),2.30(dd,J=12.0,8.5Hz,1H) ,1.97–1.87(m,1H),1.64(m,1H),1.40(t,J=7.0Hz,3H),1.35(d,J=11.3Hz,1H),1.23(s,1H); ESI-MS:m / z=355[M+H] + .

[0121] The structures of intermediate 2-1, compound 2, intermediate 3-1, and compound 3 are as follows:

[0122]

[0123] Preparation Example 2: (R)-N 1 -(1-Ethyl-1H-pyrrolo[3,2-c]pyridin-6-yl)-5-fluoro-N 3 -(piperidin-3-yl)

[0124] Benzene-1,3-diamine (Compound 2)

[0125] Step 1: Synthesis of 6-chloro-1-ethyl-1H-pyrrolo[3,2-c]pyridine (intermediate 2-1)

[0126] According to the preparation method of intermediate 1-3, replacing 2-chloro-7H-pyrrolo[3,2-c]pyridine with 6-chloro-1H-pyrrolo[2,3-d]pyrimidine yielded white solid 2-1 (63%). 1 H NMR (500MHz, CDCl3) δ8.86(d,J=1.0Hz,1H),7.28(t,J=1.0Hz,1H),7.14(d,J=3.0Hz,1H),6.5 8(dd,J=3.0,1.0Hz,1H),4.14(q,J=7.5Hz,2H),1.47(t,J=7.5Hz,3H); ESI-MS:m / z=181[M+H] + .

[0127] Step 2: (R)-N 1 -(1-Ethyl-1H-pyrrolo[3,2-c]pyridin-6-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 2)

[0128] A white solid (47%) was prepared using intermediates 1-2 and 2-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ8.56(s,1H),8.46(s,1H),7.26(d,J=3.5Hz,1H),6.83(s,1H),6.69(d,J=12.0 Hz,1H),6.51(s,1H),6.43(d,J=3.5Hz,1H),5.84(d,J=12.0Hz,1H),5.58(d,J=8.0Hz,1H),4.08(q,J=7 .0Hz,2H),3.19(s,1H),3.10(d,J=12.0Hz,1H),2.84(d,J=12.0Hz,1H),2.32(m,2H),1.92(d,J=12.5H z,1H),1.64(s,1H),1.44(d,J=12.5Hz,1H),1.34(t,J=7.0Hz,3H),1.26(m,1H); ESI-MS:m / z=354[M+H] + .

[0129] Preparation Example 3: (R)-N 1 -(1-Ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-yl)-5-fluoro-N 3 -(pyridine-3-yl)phenyl-1,3-diamine (compound 3)

[0130] Step 1: Synthesis of 6-chloro-1-ethyl-1H-pyrazolo[3,4-d]pyrimidine (intermediate 3-1)

[0131] According to the preparation method of intermediate 1-3, replacing 2-chloro-7H-pyrrolo[2,3-d]pyrimidine with 6-chloro-1H-pyrazolo[3,4-d]pyrimidine yields pale yellow solid 3-1 (63%). 1 H NMR (500MHz, CDCl3) δ9.03 (s, 1H), 8.15 (s, 1H), 4.53 (q, J = 7.5Hz, 2H), 1.54 (t, J = 7.5Hz, 3H); ESI-MS: m / z = 183 [M+H] + .

[0132] Step 2: (R)-N 1-(1-Ethyl-1H-pyrazolo[3,4-d]pyrimidin-6-yl)-5-fluoro-N 3 Synthesis of 1,3-(pyridine-3-yl)phenyl-1,3-diamine (compound 3)

[0133] A white solid (46%) was prepared using intermediates 1-2 and 3-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, CDCl3) δ8.81(s,1H),7.91(s,1H),7.31(s,1H),7.07(dt,J=11.0,2.0Hz,1H),6.68(d,J=2 .0Hz,1H),6.06(dt,J=11.0,2.0Hz,1H),4.43(q,J=7.0Hz,2H),4.19(s,1H),3.50(m,1H),3.27–3.21(m ,1H),2.91(d,J=8.5Hz,1H),2.79(d,J=10.0Hz,1H),2.67(dd,J=12.0,7.0Hz,1H),2.22(t,J=7.0Hz,1H ),2.01(d,J=6.5Hz,1H),1.91(m,1H),1.82–1.77(m,1H),1.54(t,J=7.0Hz,3H).ESI-MS:m / z=356[M+H] + .

[0134] The structures of intermediate 4-1, compound 4, intermediate 5-1, and compound 5 are as follows:

[0135]

[0136] Preparation Example 4: (R)-N 1 -(9-Ethyl-9H-purin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 4)

[0137] Step 1: Synthesis of 2-chloro-9-ethyl-9H-purine (intermediate 4-1)

[0138] According to the preparation method of intermediate 1-3, replacing 2-chloro-7H-pyrrolo[2,3-d]pyrimidine with 2-chloro-9H-purine yields pale yellow solid 4-1 (57%). 1 H NMR (500MHz, CDCl3) δ9.00 (s, 1H), 8.13 (s, 1H), 4.36 (t, J = 14.5Hz, 2H), 1.60 (d, J = 14.5Hz, 3H); ESI-MS: m / z = 183 [M+H] + .

[0139] Step 2: (R)-N 1 -(9-Ethyl-9H-purin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 4)

[0140] A white solid (37%) was prepared from intermediates 1-2 and 4-1 according to step 4 of Example 1. ¹H NMR (500 MHz, CDCl₃) δ 8.80 (s, ¹H), 7.84 (s, ¹H), 7.39 (s, ¹H), 7.07 (dt, J = 11.0, 2.0 Hz, ¹H), 6.63 (t, J = 2.0 Hz, ¹H), 6.03 (dt, J = 11.0, 2.0 Hz, ¹H), 4.24 (q, J = 7.5 Hz, 2H), 3.55–3.46 (m, ¹H), 3.25 (dd, J = 12 Hz, 2H). .0,3.5Hz,1H),2.99–2.89(m,2H),2.80(td,J=8.0,3.5Hz,1H),2.68(dd,J=12.0,7.0Hz,1H),1.91(d ,J=11.0Hz,1H),1.86–1.78(m,1H),1.57(t,J=7.5Hz,3H),1.27–1.25(m,2H); ESI-MS:m / z=356[M+H] + .

[0141] Preparation Example 5: (R)-N 1 -(5-ethyl-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 5)

[0142] Step 1: Synthesis of 2-chloro-5-ethyl-5H-pyrrolo[3,2-d]pyrimidine (intermediate 5-1)

[0143] According to the preparation method of intermediate 1-3, replacing 2-chloro-7H-pyrrolo[3,2-d]pyrimidine with 2-chloro-7H-pyrrolo[2,3-d]pyrimidine yields white solid 5-1 (75%). 1 H NMR (500MHz, CDCl3) δ8.47(s,1H),7.41(d,J=4.5Hz,1H),6.24(d,J=4.5Hz,1H),4.25(q,J=7.5Hz,2H),1.39(t,J=7.5Hz,3H); ESI-MS: m / z=182[M+H] + .

[0144] Step 2: (R)-N 1-(5-ethyl-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 5)

[0145] A white solid (61%) was prepared using intermediates 1-2 and 5-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.11(s,1H),8.81(s,1H),7.73(d,J=3.0Hz,1H),7.05(d,J=12.0Hz,1H),6.92(s,1 H),6.32(d,J=3.0Hz,1H),5.92(d,J=12.0Hz,1H),5.58(d,J=8.0Hz,1H),4.24(q,J=7.0Hz,2H),3.21(m,1H) ,3.11(d,J=12.0Hz,1H),2.82(d,J=12.0Hz,1H),2.47(d,J=11.0Hz,1H),2.39-2.28(m,1H),2.00–1.90(m, 1H),1.66(m,1H),1.48-1.42(m,1H),1.41–1.38(t,J=7.0Hz,3H),1.38–1.33(m,1H); ESI-MS:m / z=355[M+H] + .

[0146] The structures of intermediate 6-1, compound 6, compound 7, and compound 8 are as follows:

[0147]

[0148] Preparation Example 6: (R)-N 1 -(1-Ethyl-1H-pyrrolo[2,3-c]pyridin-5-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 6)

[0149] Step 1: Synthesis of 5-chloro-1-ethyl-1H-pyrrolo[2,3-c]pyridine (intermediate 6-1)

[0150] According to the preparation method of intermediate 1-3, replacing 2-chloro-7H-pyrrolo[2,3-c]pyridine with 5-chloro-1H-pyrrolo[2,3-d]pyrimidine yields white solid 6-1 (45%). 1H NMR (500MHz, CDCl3) δ8.76 (s, 1H), 7.53 (t, J = 1.0Hz, 1H), 7.21 (d, J = 3.0Hz, 1H), 6.33 (dd ,J=3.0,1.0Hz,1H),4.24(q,J=8.0Hz,2H),1.23(t,J=8.0Hz,3H); ESI-MS:m / z=181[M+H] + .

[0151] Step 2: (R)-N 1 -(1-Ethyl-1H-pyrrolo[2,3-c]pyridin-5-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 6)

[0152] A white solid (39%) was prepared using intermediates 1-2 and 6-1 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ8.54(s,1H),8.39(s,1H),7.51(d,J=3.0Hz,1H),7.04–6.97(m,1H),6.64(dt,J=12.0,2.0Hz, 1H),6.44(d,J=2.0Hz,1H),6.31(d,J=3.0Hz,1H),5.82(dt,J=12.0,2.0Hz,1H),5.51(d,J=8.0Hz,1H),4.23(q,J=7.0H z,2H),3.15(dd,J=8.0,4.0Hz,1H),3.06(s,1H),2.79(d,J=12.0Hz,1H),2.44(t,J=10.5Hz,1H),2.28(t,J=10.0Hz,1H ),1.95–1.86(m,1H),1.63(m,1H),1.46–1.40(m,1H),1.38(t,J=7.0Hz,3H),1.36–1.28(m,1H); ESI-MS:m / z=354[M+H] + .

[0153] Preparation Example 7: (R)-5-fluoro-N 1 -(piperidin-3-yl)-N 3 -(quinazolin-2-yl)phenyl-1,3-diamine (compound 7)

[0154] Step 1: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 Synthesis of 1,3-(quinazolin-2-yl)phenyl-1,3-diamine (compound 7)

[0155] A pale yellow solid (62%) was prepared using intermediates 1-2 and 2-chloroquinazoline as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.78(s,1H),9.30(s,1H),7.92(dd,J=8.0,1.5Hz,1H),7.83(m,1H),7.69(dd,J =8.0,1.0Hz,1H),7.40(m,1H),7.23(d,J=2.0Hz,1H),7.03(m,1H),6.03(m,1H),5.72(d,J=8.0Hz,1H), 3.21(d,J=4.0Hz,1H),3.14(d,J=12.0Hz,1H),2.83(d,J=12.0Hz,1H),2.47(t,J=11.0Hz,1H),2.32(dd ,J=12.0,8.5Hz,1H),2.02–1.93(m,1H),1.66(m,1H),1.45(m,1H),1.37(m,1H).ESI-MS:m / z=338[M+H] + .

[0156] Preparation Example 8: (R)-5-fluoro-N 1 -(isoquinoline-3-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 8)

[0157] Step 1: (R)-5-Fluoro-N 1 -(isoquinoline-3-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 8)

[0158] A pale yellow solid (52%) was prepared using intermediates 1-2 and 3-chloroisoquinoline as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.04(s,1H),8.95(s,1H),7.92(d,J=8.0Hz,1H),7.70(d,J=8.0Hz,1H),7.57(t,J=7. 5Hz,1H),7.32(t,J=7.5Hz,1H),7.17(s,1H),6.64(d,J=12.0Hz,1H),6.55(s,1H),5.95(d,J=12.0Hz,1H),5. 68(d,J=8.0Hz,1H),3.19(m,1H),3.10(d,J=12.0Hz,1H),2.80(d,J=12.0Hz,1H),2.45(t,J=11.0Hz,1H),2.3 5–2.25(m,1H),2.02–1.88(m,1H),1.64(m,1H),1.48–1.39(m,1H),1.37–1.29(m,1H); ESI-MS: m / z=337[M+H] + .

[0159] The structures of compound 9, intermediate 10-1, compound 10, intermediate 11-1, and compound 11 are as follows:

[0160]

[0161] Preparation Example 9: (R)-5-fluoro-N 1 -(piperidin-3-yl)-N 3 -(7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 9)

[0162] Step 1: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 Synthesis of 7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 9)

[0163] A white solid (23%) was prepared from intermediates 1-2 and 2-chloro-7H-pyrrolo[2,3-d]pyrimidine according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ11.52(s,1H),9.19(s,1H),8.68(s,1H),7.18(d,J=3.5Hz,1H),7.15(m,1H),6.71 (t,J=2.0Hz,1H),6.40(d,J=3.5Hz,1H),5.93(dt,J=12.0,2.0Hz,1H),5.56(d,J=8.0Hz,1H),3.20(d,J=8 .5Hz,1H),3.09(d,J=12.0Hz,1H),2.81(d,J=12.0Hz,1H),2.46(d,J=11.0Hz,1H),2.31(dd,J=12.0,8.5H z,1H),1.91(d,J=12.0Hz,1H),1.68–1.60(m,1H),1.42(m,1H),1.38–1.30(m,1H); ESI-MS:m / z=327[M+H] + .

[0164] Preparation Example 10: (R)-5-Fluoro-N 1 -(7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 10)

[0165] Step 1: Synthesis of 2-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 10-1)

[0166] According to the preparation method of intermediate 1-3, iodomethane was used instead of iodoethane to obtain white solid 10-1 (77%). 1 HNMR (500MHz, CDCl3) δ8.77 (s, 1H), 7.31 (d, J = 3.5Hz, 1H), 6.10 (d, J = 3.5Hz, 1H), 3.62 (s, 3H); ESI-MS: m / z = 168 [M+H] + .

[0167] Step 2: (R)-5-Fluoro-N 1 -(7-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 10)

[0168] A white solid (60%) was prepared using intermediates 1-2 and 10-1 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.31 (s, 1H), 8.67 (s, 1H), 7.23 (d, J = 3.5Hz, 1H), 7.08 (s, 1H), 6.93 (d ,J=12.0Hz,1H),6.43(d,J=3.5Hz,1H),5.95(d,J=12.0Hz,1H),5.60(d,J=8.0Hz,1H),3.74(s, 3H),3.23–3.19(m,1H),3.11(d,J=12.0Hz,1H),2.80(d,J=12.0Hz,1H),2.48–2.41(m,1H),2.2 9(t,J=10.0Hz,1H),1.99–1.89(m,1H),1.64(m,1H),1.49–1.30(m,2H); ESI-MS:m / z=341[M+H] + .

[0169] Preparation Example 11: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 -(7-propyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 11)

[0170] Step 1: Synthesis of 2-chloro-7-propyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 11-1)

[0171] According to the preparation method of intermediate 1-3, replacing iodoethane with 1-iodopropane yields white solid 11-1 (82%). 1 HNMR(500MHz, CDCl3)δ8.78(s,1H),7.31(d,J=3.5Hz,1H),6.52(d,J=3.5Hz,1H), 4.17(t,J=7.0Hz,2H),1.49(m,2H); 0.89(t,J=8.0Hz,3H); ESI-MS:m / z=196[M+H] + .

[0172] Step 2: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 Synthesis of 1,3-(7-propyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 11)

[0173] A white solid (61%) was prepared using intermediates 1-2 and 11-1 as raw materials according to step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.29(s,1H),8.67(s,1H),7.28(d,J=3.5Hz,1H),7.00(dt,J=14.5,2.0Hz,2H),6 .43(d,J=3.5Hz,1H),5.96(dt,J=12.0,2.0Hz,1H),5.57(d,J=8.1Hz,1H),4.12(m,2H),3.12–3.05(m,1H ),2.79(dd,J=10.5,6.5Hz,1H),2.46(d,J=12.0Hz,1H),2.31(dd,J=12.0,8.5Hz,1H),1.91(m,1H),1.84 (m,2H),1.69–1.61(m,1H),1.46–1.32(m,2H),1.23(s,1H),0.85(t,J=7.5Hz,3H); ESI-MS:m / z=369[M+H] + .

[0174] The structures of intermediate 12-1, compound 12, intermediate 13-1, and compound 13 are as follows:

[0175]

[0176] Preparation Example 12: (R)-N 1 -(7-Cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 12)

[0177] Step 1: Synthesis of 2-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 12-1)

[0178] 2-Chloro-7H-pyrrolo[2,3-d]pyrimidine (100 mg, 0.65 mmol), cyclopropylboronic acid (112 mg, 1.30 mmol), and triethylamine (132 mg, 1.30 mmol) were dissolved in DCM (10 ml), and ketone acetate (118 mg, 0.65 mmol) was added. The mixture was reacted overnight at room temperature with an open container. The mixture was filtered, and the solvent was recovered under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography with PE:EA = 6:1 as the eluent to give a white solid 12-1 (92 mg, 73%). 1 H NMR (500MHz, CDCl3) δ8.78 (s, 1H), 7.31 (d, J = 3.5Hz, 1H), 6.52 (d, J = 3.5Hz, 1H), 2.56 (m, 1H), 1.03 (m, 4H); ESI-MS: m / z = 194 [M+H] + .

[0179] Step 2: (R)-N 1 -(7-Cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 12)

[0180] A white solid (56%) was prepared using intermediates 1-2 and 12-1 as raw materials according to the method in step 4 of Example 1. 1 H NMR (500MHz, CDCl3) δ8.58 (s, 1H), 7.30 (s, 1H), 7.22 (s, 1H), 6.93 (d, J = 3.5Hz, 1H), 6.62 (s, 1H),6.33(d,J=3.5Hz,1H),6.01(d,J=11.0Hz,1H),3.52(s,1H),3.41(dd,J=8.0,4.0Hz,1H) ,3.24(d,J=12.0Hz,1H),2.92(m,1H),2.79(m,1H),2.66(m,1H),1.92(m,1H),1.81(m,1H),1 .58(m,1H)1.25(m,2H),1.14(d,J=7.5Hz,2H),1.08(d,J=4.5Hz,2H); ESI-MS:m / z=367[M+H] + .

[0181] Preparation Example 13: (R)-5-Fluoro-N 1 -(7-Isopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 13)

[0182] Step 1: Synthesis of 2-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 13-1)

[0183] Following the preparation method of intermediate 12-1, replacing cyclopropylboronic acid with isopropylboronic acid yielded white solid 13-1 (75%). ESI-MS: m / z = 196 [M+H] + .

[0184] Step 2: (R)-5-Fluoro-N 1 -(7-Isopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 13)

[0185] A white solid (47%) was prepared using intermediates 1-2 and 13-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, CDCl3) δ8.61(s,1H),7.17(s,1H),7.10(dt,J=11.0,2.0Hz,1H),7.04(d,J=3.5Hz,1H),6.64(d,J= 2.0Hz,1H),6.40(d,J=3.5Hz,1H),6.00(dt,J=11.0,2.0Hz,1H),5.00(m,1H),4.11(s,1H),3.47(m,1H),3.22(d d,J=12.0,3.5Hz,1H),2.90(dd,J=13.5,6.0Hz,1H),2.80–2.72(m,1H),2.63(dd,J=12.0,7.0Hz,1H),2.49(m,1 H),1.85–1.73(m,1H),1.57(d,J=9.5Hz,1H),1.53(d,J=7.0Hz,6H),1.28–1.23(m,1H); ESI-MS: m / z=369[M+H]+.

[0186] The structures of intermediate 14-1, compound 14, intermediate 15-1, and compound 15 are as follows:

[0187]

[0188] Preparation Example 14: (R)-N 1 -(7-cyclohexyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 14)

[0189] Step 1: Synthesis of 2-chloro-7-cyclohexyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 14-1)

[0190] Following the preparation method for intermediates 1-3, replacing iodoethane with iodocyclohexane yields white solid 14-1 (26%). ESI-MS: m / z = 236 [M+H] + .

[0191] Step 2: (R)-N 1 -(7-cyclohexyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 14)

[0192] A white solid (38%) was prepared using intermediates 1-2 and 14-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.28(s,1H),8.66(s,1H),7.36(d,J=3.5Hz,1H),7.11(d,J=12.5Hz,1H),6.82(t,J=2.0Hz, 1H),6.42(d,J=3.5Hz,1H),6.01–5.92(m,1H),5.57(d,J=8.0Hz,1H),4.47(t,J=12.0Hz,1H),3.51(s,1H),3.24(s, 1H),3.08(d,J=11.5Hz,1H),2.80(d,J=12.0Hz,1H),2.33(d,J=11.0Hz,1H),1.94(s,2H),1.87(d,J=12.0Hz,4H),1 .72(d,J=13.0Hz,1H),1.64(s,1H),1.53–1.31(m,4H),1.29(d,J=13.5Hz,1H),1.23(s,2H).ESI-MS:m / z=409[M+H] + .

[0193] Preparation Example 15: (R)-5-Fluoro-N 1 -(7-(oxecyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 15)

[0194] Step 1: Synthesis of 2-chloro-7-(oxetane-3-yl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 15-1)

[0195] Following the preparation method for intermediates 1-3, replacing iodoethane with 3-iodooxetane yields white solid 15-1 (75%). ESI-MS: m / z = 210 [M+H] + .

[0196] Step 2: (R)-5-Fluoro-N 1 -(7-(oxecyclobutane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 15)

[0197] A white solid (52%) was prepared using intermediates 1-2 and 15-1 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, CDCl3) δ8.57(s,1H),7.37(s,1H),7.15(d,J=4.0Hz,1H),7.08(t,J=2.0Hz,1H),6.68(dt,J=11.0,2 .0Hz,1H),6.41(d,J=4.0Hz,1H),5.93(dt,J=11.02.0Hz,1H),5.71(m,1H),5.20(td,J=6.5Hz,2H),5.09(td,J=7 .5,3.0Hz,2H),4.32(s,1H),3.40(m,1H),3.19(dd,J=12.0,3.5Hz,1H),2.93(s,1H),2.87(dt,J=11.0,3.0Hz,1H ),2.69(m,1H),2.55(dd,J=12.0,7.5Hz,1H),1.87(m,1H),1.72(m,1H),1.49–1.42(m,1H); ESI-MS:m / z=383[M+H] + .

[0198] The structures of intermediate 16-1, compound 16, intermediate 17-1, and compound 17 are as follows:

[0199]

[0200] Preparation Example 16: (R)-1-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylprop-2-ol (Compound 16)

[0201] Step 1: Synthesis of 1-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylprop-2-ol (intermediate 16-1)

[0202] Following the preparation method of intermediates 1-3, 2,2-dimethylethylene oxide was used instead of iodoethane, and triethylamine was used instead of NaH to obtain oily liquid 16-1 (54%). ESI-MS: m / z = 226 [M+H] + .

[0203] Step 2: Synthesis of (R)-1-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylprop-2-ol (compound 16)

[0204] A white solid (49%) was prepared using intermediates 1-2 and 16-1 as raw materials according to step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.29(s,1H),8.68(s,1H),7.25(d,J=3.5Hz,1H),7.03–6.96(m,2H),6.4 3(d,J=3.5Hz,1H),5.96(m,1H),5.56(d,J=8.0Hz,1H),4.10(s,2H),3.25(m,1H),3.11(d,J=12.0 Hz,1H),2.79(d,J=12.0Hz,1H),2.46(t,J=11.0Hz,1H),2.34–2.26(m,1H),1.91(d,J=12.0Hz,1H ),1.64(m,1H),1.45(m,1H),1.40–1.32(m,1H),1.11(s,3H),1.10(s,3H).ESI-MS: m / z=399[M+H] + .

[0205] Preparation Example 17: (R)-5-Fluoro-N 1 -(7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 17)

[0206] Step 1: Synthesis of 2-chloro-7-(methanesulfonyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 17-1)

[0207] 2-Chloro-7H-pyrrolo[2,3-d]pyrimidine (100 mg, 0.65 mmol) and triethylamine (99 mg, 0.98 mmol) were dissolved in DCM (2 ml). A DCM solution (1 M) of methanesulfonyl chloride (89 mg, 0.78 mmol) was slowly added dropwise under ice bath. After the addition was complete, the mixture was transferred to room temperature and reacted for 1 h. The solvent was recovered by vacuum distillation to obtain the residue, which was purified by silica gel column chromatography with PE:EA = 1:1 as the eluent to give a white solid 17-1 (134 mg, 89%). 1 H NMR (500MHz, CDCl3) δ8.67 (s, 1H), 7.32 (d, J = 3.5Hz, 1H), 6.33 (d, J = 3.5Hz, 1H), 3.13 (s, 3H); ESI-MS: m / z = 232 [M + H] +.

[0208] Step 2: (R)-5-Fluoro-N 1 -(7-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 17)

[0209] A white solid (50%) was prepared using intermediates 1-2 and 17-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, CDCl3) δ8.69(s,1H),7.39(s,1H),7.34(d,J=4.0Hz,1H),6.90(d,J=2.0Hz,1 H),6.79(dt,J=11.0,2.0Hz,1H),6.54(d,J=4.0Hz,1H),6.03(dt,J=11.0,2.0Hz,1H),4.1 7(s,1H),3.54(s,3H),3.24(d,J=12.0Hz,1H),2.95–2.90(m,1H),2.82–2.73(m,1H),2.63 (t,J=10.0Hz,1H),2.52(m,1H),1.97–1.89(m,1H),1.83–1.74(m,1H),1.62–1.54(m,2H). ESI-MS:m / z=405[M+H] + .

[0210] The structures of intermediate 18-1, compound 18, intermediate 19-1, and compound 19 are as follows:

[0211]

[0212] Preparation Example 18: (R)-5-Fluoro-N 1 -(7-Phenyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 18)

[0213] Step 1: Synthesis of 2-chloro-7-phenyl-7H-pyrrolo[2,3-d]pyrimidine (intermediate 18-1)

[0214] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with phenylboronic acid yielded a white solid 18-1 (89%). ESI-MS: m / z = 230 [M+H] + .

[0215] Step 2: (R)-5-Fluoro-N 1 -(7-Phenyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 18)

[0216] A white solid (54%) was prepared using intermediates 1-2 and 18-1 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, CDCl3) δ8.71(s,1H),7.79(d,J=8.0Hz,2H),7.54(q,J=8.0Hz,2H),7.36(t,J=7.5Hz ,1H),7.32(s,1H),7.28(s,1H),6.93(dt,J=11.5,2.0Hz,1H),6.78(s,1H),6.57(d,J=4.0Hz,1H) ,5.98(dt,J=11.5,2.0Hz,1H),4.05(s,1H),3.36(m,1H),3.16(dd,J=12.0,3.5Hz,1H),2.92–2.8 1(m,1H),2.73(m,1H),2.57(m,1H),1.86(m,1H),1.73(m,1H),1.50(m,2H); ESI-MS:m / z=403[M+H] + .

[0217] Preparation Example 19: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 -(7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 19)

[0218] Step 1: Synthesis of 2-chloro-7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 19-1)

[0219] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with pyridine-4-boric acid yielded white solid 19-1 (82%). ESI-MS: m / z = 231 [M+H] + .

[0220] Step 2: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 Synthesis of 7-(pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 19)

[0221] A white solid (41%) was prepared using intermediates 1-2 and 19-1 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.57(s,1H),8.84(s,1H),8.70(d,J=2.0Hz,1H),8.69(d,J=2.0Hz,1H),8.19(d,J=2.0Hz,1H) ,8.18(d,J=2.0Hz,1H),7.90(d,J=4.0Hz,1H),7.06(dt,J=12.0,2.0Hz,1H),6.81–6.73(m,2H),6.02(dt,J=12.0,2.0H z,1H),5.63(d,J=8.0Hz,1H),3.19(m,1H),3.06(dd,J=12.5,3.5Hz,1H),2.78(dt,J=12.5,4.0Hz,1H),2.49-2.42(m,1 H),2.31(dd,J=12.0,8.5Hz,1H),1.90(d,J=10.5Hz,1H),1.68–1.57(m,1H),1.44–1.29(m,2H); ESI-MS:m / z=404[M+H] + .

[0222] The structures of intermediate 20-1, compound 20, intermediate 21-1, and compound 21 are as follows:

[0223]

[0224] Preparation Example 20: (R)-5-Fluoro-N 1 -(7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 20)

[0225] Step 1: Synthesis of 2-chloro-7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 20-1)

[0226] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with (1-methyl-1H-pyrazol-4-yl)boronic acid yielded a white solid 20-1 (74%). ESI-MS: m / z = 234 [M+H] + .

[0227] Step 2: (R)-5-Fluoro-N 1 -(7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 20)

[0228] A white solid (43%) was prepared using intermediates 1-2 and 20-1 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.43(s,1H),8.77(s,1H),8.38(s,1H),8.05(s,1H),7.58(d,J=4.0Hz,1H),7.08( dt,J=12.0,2.0Hz,1H),6.77(s,1H),6.63(d,J=4.0Hz,1H),6.01(dt,J=12.0,2.0Hz,1H),5.67(d,J=8.0Hz ,1H),3.93(s,3H),3.27–3.18(m,1H),3.10(d,J=12.0Hz,1H),2.83(d,J=13.0Hz,1H),2.48(s,1H),2.34( t,J=10.5Hz,1H),1.92(d,J=13.0Hz,1H),1.66(t,J=9.0Hz,1H),1.49–1.30(m,2H); ESI-MS:m / z=407[M+H] + .

[0229] Preparation Example 21: (R)-5-Fluoro-N 1 -(7-(2-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 21)

[0230] Step 1: Synthesis of 2-chloro-7-(2-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 21-1)

[0231] Following the preparation method of intermediate 12-1, replacing cyclopropylboronic acid with 2-fluorophenylboronic acid yielded white solid 21-1 (23%). ESI-MS: m / z = 248 [M+H] + .

[0232] Step 2: (R)-5-Fluoro-N 1 -(7-(2-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 21)

[0233] A white solid (40%) was prepared using intermediates 1-2 and 21-1 as raw materials according to step 4 of Example 1. 1H NMR(500MHz,DMSO-d6)δ9.39(s,1H),8.81(s,1H),7.74(t,J=7.0Hz,1H),7.59–7.49(m,2H),7.46(d,J=3.5Hz ,1H),7.42(t,J=7.0Hz,1H),7.00(d,J=12.0Hz,1H),6.75–6.65(m,2H),5.98–5.89(m,1H),5.45(d,J=8.0Hz,1 H),3.12(m,1H),3.00(dd,J=12.0,3.5Hz,1H),2.76(dd,J=11.0,6.5Hz,1H),2.48(d,J=8.0Hz,1H),2.31(dd, J=12.0,8.0Hz,1H),1.82(d,J=11.0Hz,1H),1.60(dd,J=12.0,6.0Hz,1H),1.37(m,2H); ESI-MS: m / z=421[M+H] + .

[0234] The structures of intermediate 22-1, compound 22, intermediate 23-1, and compound 23 are as follows:

[0235]

[0236] Preparation Example 22: (R)-5-Fluoro-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 22)

[0237] Step 1: Synthesis of 2-chloro-7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 22-1)

[0238] According to the preparation method of intermediate 12-1, 3-fluorophenylboronic acid was used instead of cyclopropylboronic acid to obtain white solid 22-1 (77%). 1 H NMR (500MHz, CDCl3) δ8.90 (s, 1H), 7.55–7.49 (m, 4H), 7.11 (m, 1H), 6.76 (d, J = 4.0Hz, 1H); ESI-MS: m / z = 248 [M+H] + .

[0239] Step 2: (R)-5-Fluoro-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 22)

[0240] A white solid (60%) was prepared using intermediates 1-2 and 22-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.49(s,1H),8.82(s,1H),7.94(dt,J=11.0,2.5Hz,1H),7.82(dd,J=8.0,2.0Hz,1H),7.75(d,J=4. 0Hz,1H),7.60(td,J=8.0,6.5Hz,1H),7.23(td,J=8.5,2.5Hz,1H),7.11(dt,J=12.0,2.0Hz,1H),6.71(t,J=2.5Hz,2H),5.9 9(dt,J=12.0,2.0Hz,1H),5.52(d,J=8.0Hz,1H),3.16(m,1H),3.04(dd,J=12.0,4.0Hz,1H),2.77(dt,J=12.0,4.0Hz,1H), 2.46(m,1H),2.30(dd,J=12.0,8.5Hz,1H),1.87(dd,J=9.5,5.0Hz,1H),1.62(m,1H),1.37(m,2H); ESI-MS: m / z=421[M+H]+.

[0241] Preparation Example 23: (R)-5-Fluoro-N 1 -(7-(4-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 23)

[0242] Step 1: Synthesis of 2-chloro-7-(4-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 23-1)

[0243] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with 4-fluorophenylboronic acid yielded white solid 23-1 (84%). ESI-MS: m / z = 248 [M+H] + .

[0244] Step 2: (R)-5-Fluoro-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 23)

[0245] A white solid (53%) was prepared using intermediates 1-2 and 23-1 as raw materials according to step 4 of Example 1.1 H NMR(500MHz,DMSO-d6)δ9.44(s,1H),8.81(s,1H),7.94-7.89(m,2H),7.63(d,J=4.0Hz,1H),7.44–7.39(m,2H ),7.04(dt,J=12.0,2.0Hz,1H),6.77(d,J=2.0Hz,1H),6.68(d,J=4.0Hz,1H),5.97(dt,J=12.0,2.0Hz,1H),5 .51(d,J=8.0Hz,1H),3.15(m,1H),3.05–2.97(m,1H),2.76(dt,J=13.0,4.0Hz,1H),2.50–2.44(m,1H),2.32( dd,J=12.0,8.0Hz,1H),1.89–1.80(m,1H),1.62(dd,J=13.0,4.0Hz,1H),1.36(m,2H); ESI-MS:m / z=421[M+H] + .

[0246] The structures of intermediate 24-1, compound 24, intermediate 25-1, and compound 25 are as follows:

[0247]

[0248] Preparation Example 24: (R)-N 1 -(7-(2-chloro-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 24)

[0249] Step 1: Synthesis of 2-chloro-7-(2-chloro-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 24-1)

[0250] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with 2-chloro-3-fluorophenylboronic acid yielded a white solid 24-1 (19%). ESI-MS: m / z = 282 [M+H] + .

[0251] Step 2: (R)-N 1 -(7-(2-chloro-3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-5-fluoro-N 3 Synthesis of 1,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 24)

[0252] A white solid (29%) was prepared using intermediates 1-2 and 24-1 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz, CDCl3)δ8.72(s,1H),7.46–7.40(m,2H),7.32–7.27(m,1H),7.15(d,J=3.5Hz,1H),7.13(s,1H) ,6.77(dt,J=11.5,2.0Hz,1H),6.69(d,J=2.0Hz,1H),6.60(d,J=3.5Hz,1H),5.96(dt,J=11.0,2.0Hz,1H),3. 99(s,1H),3.33(m,1H),3.15(dd,J=12.0,3.0Hz,1H),2.86(d,J=9.0Hz,1H),2.75(d,J=9.0Hz,1H),2.58(dd, J=11.5,7.5Hz,1H),1.84(d,J=11.0Hz,1H),1.79-1.70(m,1H),1.52(t,J=8.5Hz,2H); ESI-MS:m / z=455[M+H] + .

[0253] Preparation Example 25: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 -(7-(3-methylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 25)

[0254] Step 1: Synthesis of 2-chloro-7-(3-methylphenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 25-1)

[0255] According to the preparation method of intermediate 12-1, 3-methylphenylboronic acid was used instead of cyclopropylboronic acid to obtain white solid 25-1 (72%). 1 H NMR (500MHz, CDCl3) δ8.88(s,1H),7.49(d,J=3.5Hz,1H),7.47(d,J=5.0Hz,2H),7.42(t,J=7 .5Hz,1H),7.22(d,J=7.5Hz,1H),6.72(d,J=3.5Hz,1H),2.46(s,3H); ESI-MS:m / z=244[M+H] + .

[0256] Step 2: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3Synthesis of 7-(3-methylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 25)

[0257] A white solid (44%) was prepared using intermediates 1-2 and 25-1 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz, CDCl3)δ8.70(s,1H),7.64(t,J=2.0Hz,1H),7.57–7.53(m,1H),7.43(s,1H),7.27(d,J=3.5Hz,1H),7 .18(d,J=7.0Hz,2H),7.07(dt,J=11.5,2.0Hz,1H),6.61(t,J=2.0Hz,1H),6.56(d,J=4.0Hz,1H),5.98(dt,J=11.0, 2.0Hz,1H),3.99(d,J=8.0Hz,1H),3.36(dd,J=8.0,4.0Hz,1H),3.24–3.13(m,1H),2.91–2.83(m,1H),2.72(dd,J=1 2.0,8.0Hz,1H),2.56(dd,J=12.0,7.0Hz,1H),2.48(s,3H),1.74(m,1H),1.56–1.45(m,2H).ESI-MS:m / z=417[M+H] + .

[0258] The structures of intermediate 26-1, compound 26, intermediate 27-1, and compound 27 are as follows:

[0259]

[0260] Preparation Example 26: (R)-5-Fluoro-N 1 -(7-(3-methoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 26)

[0261] Step 1: Synthesis of 2-chloro-7-(3-methoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 26-1)

[0262] Following the preparation method of intermediate 12-1, replacing cyclopropylboronic acid with 3-methoxyphenylboronic acid yielded a white solid 26-1 (72%). ¹H NMR (500 MHz, CDCl₃) δ 8.89 (s, ¹H), 7.52 (d, J = 4.0 Hz, ¹H), 7.44 (t, J = 8.0 Hz, ¹H), 7.30 (t, J = 2.5 Hz, ¹H), 7.27 (d, J = 4.5 Hz, ¹H), 6.95 (dd, J = 8.0, 2.5 Hz, 1H), 6.72 (d, J = 4.0 Hz, 1H), 3.89 (s, ³H); m / z = 260 [M+H] + .

[0263] Step 2: (R)-5-Fluoro-N 1 Synthesis of -(7-(3-methoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N3-(piperidin-3-yl)phenyl-1,3-diamine (compound 26)

[0264] A white solid (39%) was prepared using intermediates 1-2 and 26-1 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz, CDCl3)δ8.70(s,1H),7.47–7.40(m,2H),7.32(ddd,J=8.0,2.0,1.0Hz,1H),7.27(d,J=4.0Hz,1H),7.19(s, 1H),6.92(ddd,J=8.0,2.5,1.0Hz,1H),6.87(dt,J=11.0,2.0Hz,1H),6.82(t,J=2.0Hz,1H),6.57(d,J=4.0Hz,1H),5.98( dt,J=11.0,2.0Hz,1H),4.01(d,J=8.0Hz,1H),3.87(s,3H),3.35(dd,J=8.0,4.0Hz,1H),3.20–3.14(m,1H),2.92–2.86(m ,1H),2.71(dd,J=12.0,8.5Hz,1H),2.55(dd,J=12.0,7.5Hz,1H),1.74(m,1H),1.58–1.41(m,2H); ESI-MS:m / z=433[M+H] + .

[0265] Preparation Example 27: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 -(7-(3-(trifluoromethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 27)

[0266] Step 1: Synthesis of 2-chloro-7-(3-(trifluoromethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidine (intermediate 27-1)

[0267] According to the preparation method of intermediate 12-1, 3-(trifluoromethyl)phenylboronic acid was used instead of cyclopropylboronic acid to obtain white solid 27-1 (85%). 1 H NMR (500MHz, CDCl3) δ8.92(s,1H),8.03(dt,J=8.0,2.0Hz,1H),7.92(d,J=2.0Hz,1 H),7.74–7.65(m,2H),7.55(d,J=4.0Hz,1H),6.79(d,J=4.0Hz,1H); m / z=298[M+H] + .

[0268] Step 2: (R)-5-Fluoro-N 1 -(piperidin-3-yl)-N 3 Synthesis of 7-(3-(trifluoromethyl)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 27)

[0269] A white solid (50%) was prepared using intermediates 1-2 and 27-1 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, CDCl3) δ8.72(s,1H),8.13(dt,J=8.0,1.5Hz,1H),7.96((t,J=2.0Hz,1H),7.69(t,J=8.0Hz,1H),7.61(d,J =8.0Hz,1H),7.35(s,1H),7.28(d,J=4.0Hz,1H),6.99(dt,J=11.0,2.0Hz,1H),6.61(d,J=4.0Hz,1H),6.55(t,J=2.0Hz,1 H),5.99(dt,J=11.0,2.0Hz,1H),4.01(m,1H),3.34(m,1H),3.16(dd,J=12.0,3.5Hz,1H),2.87(ddd,J=12.0,6.0,3.5Hz ,1H),2.71(ddd,J=12.0,8.0,3.5Hz,1H),2.55(dd,J=12.0,7.5Hz,1H),1.72(m,1H),1.50(m,2H); ESI-MS:m / z=471[M+H] + .

[0270] The structures of intermediate 28-1, compound 28-2, compound 28, intermediate 29-1, and compound 29 are as follows:

[0271]

[0272] Preparation Example 28: (R)-2-(3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)prop-2-ol (Compound 28)

[0273] Step 1: Synthesis of 1-(3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)ethyl-1-one (intermediate 28-1)

[0274] Following the preparation method of intermediate 12-1, replacing cyclopropylboronic acid with 3-acetylphenylboronic acid yields white solid 28-1 (62%). m / z = 272 [M+H] + .

[0275] Step 2: Synthesis of 2-(3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)prop-2-ol (intermediate 28-2)

[0276] Intermediate 28-1 (80 mg, 0.29 mmol) was dissolved in anhydrous THF (2 mL). Under inert gas protection, 0.6 mL of a 1 M anhydrous THF solution of methyl magnesium bromide was slowly added dropwise in an ice bath. After the addition was complete, the mixture was transferred to room temperature and reacted for 3 h. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 1:2 as the eluent to give a transparent oily substance 28-2 (75 mg, 90%). m / z = 288 [M+H] + .

[0277] Step 3: Synthesis of (R)-2-(3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)phenyl)prop-2-ol (compound 28)

[0278] A white solid (36%) was prepared using intermediates 1-2 and 28-2 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, CDCl3) δ8.69(s,1H),7.97(t,J=2.0Hz,1H),7.56(dt,J=8.0,2.0Hz,1H),7.50(t,J=8.0Hz,1H),7.46(d t,J=8.0,1.5Hz,1H),7.27(s,1H),7.23(s,1H),6.86(dt,J=11.0,2.0Hz,1H),6.75(t,J=2.0Hz,1H),6.57(d,J=4.0H z,1H),5.95(dt,J=11.0,2.0Hz,1H),4.24(m,1H),3.39(s,1H),3.10(dd,J=12.0,3.5Hz,1H),2.87(d,J=7.0Hz,1H), 2.81–2.76(m,1H),2.64(dd,J=12.0,7.0Hz,1H),1.81–1.74(m,2H),1.63(s,6H),1.50(s,2H).ESI-MS:m / z=461[M+H] + .

[0279] Preparation Example 29: (R)-3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (Compound 29)

[0280] Step 1: Synthesis of 3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (intermediate 29-1)

[0281] Following the preparation method for intermediate 12-1, replacing cyclopropylboronic acid with 3-carbamoylphenylboronic acid yielded white solid 29-1 (49%). ESI-MS: m / z = 273 [M+H] + .

[0282] Step 2: Synthesis of (R)-3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (compound 29)

[0283] A white solid (53%) was prepared using intermediates 1-2 and 29-1 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.44(s,1H),8.83(s,1H),8.23(t,J=2.0Hz,1H),8.17–8.08(s,2H),7.90(dt,J=8.0,1. 5Hz,1H),7.71(d,J=4.0Hz,1H),7.66(t,J=8.0Hz,1H),7.54(s,1H),7.02(dt,J=12.0,2.0Hz,1H),6.78(t,J=2. 0Hz,1H),6.72(d,J=3.7Hz,1H),5.95(dt,J=12.0,2.0Hz,1H),5.50(d,J=8.0Hz,1H),3.13(m,1H),3.02(d,J=10 .5Hz,1H),2.77(m,1H),2.47(m,1H),2.30(m,1H),1.86(m,1H),1.60(m,1H),1.34(m,2H); ESI-MS:m / z=446[M+H] + .

[0284] Preparation Example 30: (R)-3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-N-(3-hydroxy-3-methylbutyl)benzamide (Compound 30)

[0285]

[0286] Step 1: Synthesis of methyl 3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzoate (intermediate 30-1)

[0287] Following the preparation method of intermediate 12-1, replacing cyclopropylboronic acid with 3-methoxycarbonylphenylboronic acid yielded a white solid 30-1 (35%). ESI-MS: m / z = 288 [M+H] + .

[0288] Step 2: Synthesis of 3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzoic acid (intermediate 30-2)

[0289] Intermediate 30-1 (100 mg, 0.35 mmol) was dissolved in THF:H₂O = 1:1 (1 ml: 1 ml). 0.5 ml of a 1 M lithium hydroxide aqueous solution was added at room temperature, and the reaction was allowed to proceed for 3 h. The organic solvent was removed by vacuum distillation. The remaining aqueous phase was neutralized to pH 3 with 1 M hydrochloric acid solution, precipitating a solid. The solid was obtained by filtration and washed with a small amount of distilled water, then dried under vacuum to give a white solid 30-2 (81 mg, 85%). ESI-MS: m / z = 274 [M+H]+ .

[0290] Step 3: Synthesis of 3-(2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-N-(3-hydroxy-3-methylbutyl)benzamide (intermediate 30-3)

[0291] Intermediate 30-2 (125 mg, 0.46 mmol) was dissolved in DMF (3 mL). Under ice bath conditions, HATU (225 mg, 0.59 mmol), DIPEA (236 mg, 1.82 mmol), and 4-amino-2-methylbut-2-ol (47 mg, 0.46 mmol) were added sequentially. The mixture was stirred overnight at room temperature. The reaction solution was extracted three times with ethyl acetate and water, washed three times with saturated brine, and the organic layer was dried over anhydrous sodium sulfate. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using DCM:MeOH = 50:1 as the eluent to give a clear oily substance 30-3 (132 mg, 80%). ESI-MS: m / z = 359 [M+H] + .

[0292] Step 4: Synthesis of (R)-3-(2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-N-(3-hydroxy-3-methylbutyl)benzamide (compound 30)

[0293] A white solid (39%) was prepared using intermediates 1-2 and 30-3 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.45(s,1H),8.83(s,1H),8.52(t,J=5.5Hz,1H),8.20(t,J=2.0Hz,1H),8.11(m,1H),7.84(dt,J=8.0,1.5Hz,1H) ,7.71(d,J=4.0Hz,1H),7.66(t,J=8.0Hz,1H),7.04(dt,J=12.0,2.0Hz,1H),6.78(t,J=2.0Hz,1H),6.72(d,J=4.0Hz,1H),5.95(dt,J=12 .0,2.0Hz,1H),5.53(d,J=8.0Hz,1H),3.39–3.36(m,2H),3.14(m,1H),3.04(m,1H),2.78(d,J=12.5Hz,1H),2.50–2.43(m,1H),2.31(dd, J=12.0,8.5Hz,1H),1.86(d,J=11.0Hz,1H),1.7-1.63(m,2H),1.65-1.57(m,1H),1.44-1.28(m,2H),1.15(s,6H); ESI-MS: m / z=532[M+H] + .

[0294] Preparation Example 31: (R)-3-(4-(ethylamino)-2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (Compound 31)

[0295]

[0296] Step 1: Synthesis of 2-chloro-N-ethyl-7H-pyrrolo[2,3-d]pyrimidine-4-amine (intermediate 31-1)

[0297] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (200 mg, 1.06 mmol), DIPEA (413 mg, 3.19 mmol) were administered.

[0298] (mmol) was dissolved in acetonitrile (5 ml), and 0.3 ml of 50% ethylamine aqueous solution was added at room temperature. The reaction was allowed to proceed overnight, and the solvent was recovered under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography using PE:EA = 2:1 as the eluent to give a white solid 31-1 (162 mg, 77%). ESI-MS: m / z = 197 [M+H] + .

[0299] Step 2: Synthesis of 3-(2-chloro-4-(ethylamino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (intermediate 31-2)

[0300] Following the preparation method of intermediate 12-1, a white solid 31-2 (43%) was obtained from intermediate 31-1 and 3-carbamoylphenylboronic acid. ESI-MS: m / z = 316 [M+H] + .

[0301] Step 3: Synthesis of (R)-3-(4-(ethylamino)-2-((3-fluoro-5-(piperidin-3-ylamino)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)benzamide (compound 31)

[0302] A white solid (47%) was prepared using intermediates 1-2 and 31-2 as raw materials according to step 4 of Example 1. ESI-MS: m / z = 489 [M+H] + .

[0303] Preparation Example 32: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 32)

[0304]

[0305] Step 1: Synthesis of (R)-3-((3-nitrophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 32-1)

[0306] Following the preparation method of intermediate 1-1, replacing 1-bromo-3-fluoro-5-nitrobenzene with 3-iodonitrobenzene yielded a yellow oil 32-1 (74%). ESI-MS: m / z = 322 [M+H] + .

[0307] Step 2: Synthesis of (R)-3-((3-aminophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 32-2)

[0308] According to the preparation method of intermediate 1-2, intermediate 32-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 32-2 (74%). ESI-MS: m / z = 292 [M+H] + .

[0309] Step 3: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3Synthesis of 3,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 32)

[0310] A white solid (56%) was prepared using intermediates 22-1 and 32-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz, CDCl3)δ8.70(s,1H),7.79(dt,J=10.5,2.0Hz,1H),7.55(m,1H),7.49(td,J=8.0,6.0Hz,1H),7.31(t,J =2.0Hz,1H),7.27(s,1H),7.17(s,1H),7.10(t,J=8.0Hz,1H),7.05(m,1H),6.79(dd,J=8.0,2.0Hz,1H),6.58(d,J=4. 0Hz,1H),6.31(dd,J=8.0,2.0Hz,1H),3.88(s,1H),3.41(m,1H),3.23(dd,J=12.0,3.5Hz,1H),2.91(dt,J=12.0,4.5H z,1H),2.70(m,1H),2.54(dd,J=12.0,8.0Hz,1H),1.76(m,1H),1.57–1.44(m,2H)1.43(s,1H); ESI-MS:m / z=403[M+H] + .

[0311] Preparation Example 33: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-3-yl)phenyl-1,3-diamine (compound 33)

[0312]

[0313] Step 1: Synthesis of (S)-3-((3-nitrophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 33-1)

[0314] A yellow oily substance 33-1 (68%) was prepared from 3-iodonitrobenzene and (S)-3-aminopiperidine-1-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 322 [M+H] + .

[0315] Step 2: Synthesis of (S)-3-((3-aminophenyl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 33-2)

[0316] Following the preparation method of intermediate 1-2, intermediate 33-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 33-2 (89%). ESI-MS: m / z = 292 [M+H] + .

[0317] Step 3: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 3,3-(piperidin-3-yl)phenyl-1,3-diamine (compound 33)

[0318] A white solid (63%) was prepared using intermediates 22-1 and 33-2 as raw materials according to step 4 of Example 1. ¹H NMR (500 MHz, CDCl₃) δ 8.69 (s, ¹H), 7.78 (dt, J = 10.5, 2.0 Hz, ¹H), 7.55 (m, ¹H), 7.48 (td, J = 8.0, 6.5 Hz, ¹H), 7.31 (s, ¹H), 7.25 (d, J = 4.0 Hz, ¹H), 7.10 (t, J = 8.0 Hz, ¹H), 7.05 (m, ¹H), 6.79 (m, ¹H), 6.57 (d, J = 4.0 Hz, ¹H) ,6.33–6.29(m,1H),3.88(s,1H),3.40(m,1H),3.23(dd,J=12.0,3.5Hz,1H),2.94-2.87(m,1H),2.70(m,1H ),2.53(dd,J=12.0,7.5Hz,1H),2.07–2.02(m,1H),1.75(m,1H),1.57–1.43(m,2H); ESI-MS:m / z=403[M+H] + .

[0319] Preparation Example 34: cis-N 1 -(4-Aminocyclohexyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 34)

[0320]

[0321] Step 1: Synthesis of cis-4-((3-nitrophenyl)amino)cyclohexyl)tert-butyl carbamate (intermediate 34-1)

[0322] A yellow oily substance 34-1 (76%) was prepared from 3-iodonitrobenzene and cis-(4-aminocyclohexyl)carbamate tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 336 [M+H] + .

[0323] Step 2: Synthesis of cis-4-((3-aminophenyl)amino)cyclohexyl)carbamate tert-butyl ester (intermediate 34-2)

[0324] According to the preparation method of intermediate 1-2, intermediate 34-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 34-2 (81%). ESI-MS: m / z = 306 [M+H] + .

[0325] Step 3: cis-N 1 -(4-Aminocyclohexyl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 34)

[0326] A white solid (43%) was prepared using intermediates 22-1 and 34-2 as raw materials according to step 4 of Example 1. ESI-MS: m / z = 417 [M+H] + .

[0327] Preparation Example 35: trans-N 1 -(4-Aminocyclohexyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 35)

[0328]

[0329] Step 1: Synthesis of trans-4-((3-nitrophenyl)amino)cyclohexyl)tert-butyl carbamate (intermediate 35-1)

[0330] A yellow oily substance 35-1 (67%) was prepared from 3-iodonitrobenzene and trans-(4-aminocyclohexyl)carbamate tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 336 [M+H] + .

[0331] Step 2: Synthesis of trans-4-((3-aminophenyl)amino)cyclohexyl)carbamate tert-butyl ester (intermediate 35-2)

[0332] According to the preparation method of intermediate 1-2, intermediate 35-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 35-2 (85%). ESI-MS: m / z = 306 [M+H] + .

[0333] Step 3: Trans-N 1 -(4-Aminocyclohexyl)-N 3Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 35)

[0334] A white solid (37%) was prepared using intermediates 22-1 and 35-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.25(s,1H),8.78(s,1H),8.01(dt,J=11.0,2.5Hz,1H),7.84(m,1H),7.73(d,J=4 .0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.24–7.17(m,1H),7.08–7.04(m,1H),6.96(t,J=2.0Hz,1H),6.93 (t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.20–6.15(m,1H),5.05(d,J=8.0Hz,1H),3.10–2.97(m,1H),2.5 4(dd,J=7.5,3.0Hz,1H),2.03–1.89(m,2H),1.79–1.70(m,2H),1.18–1.00(m,4H); ESI-MS:m / z=417[M+H] + .

[0335] Preparation Example 36: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(pyrrolidine-3-yl)phenyl-1,3-diamine (compound 36)

[0336]

[0337] Step 1: Synthesis of (R)-3-((3-nitrophenyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 36-1)

[0338] A yellow oily substance 36-1 (79%) was prepared from 3-iodonitrobenzene and (R)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 308 [M+H] + .

[0339] Step 2: Synthesis of (R)-3-((3-aminophenyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 36-2)

[0340] According to the preparation method of intermediate 1-2, intermediate 36-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 36-2 (91%). ESI-MS: m / z = 278 [M+H] + .

[0341] Step 3: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 3,3-(pyrrolidine-3-yl)phenyl-1,3-diamine (compound 36)

[0342] A white solid (56%) was prepared using intermediates 22-1 and 36-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.29(s,1H),8.79(s,1H),7.96(dt,J=11.0,2.5Hz,1H),7.81(dd,J=8.0,2.0Hz,1H ),7.71(d,J=4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.10–7.02(m,2H),6.95( t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.19(dd,J=8.0,2.0Hz,1H),5.41(d,J=6.5Hz,1H),3.70(m,1H),2.9 3–2.84(m,2H),2.71(m,1H),2.62(dd,J=11.5,4.0Hz,1H),1.91(m,1H),1.53(m,1H); ESI-MS:m / z=389[M+H] + .

[0343] Preparation Example 37: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(pyrrolidine-3-yl)phenyl-1,3-diamine (compound 37)

[0344]

[0345] Step 1: Synthesis of (S)-3-((3-nitrophenyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 37-1)

[0346] A yellow oily substance 37-1 (75%) was prepared from 3-iodonitrobenzene and (S)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 308 [M+H] + .

[0347] Step 2: Synthesis of (S)-3-((3-aminophenyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 37-2)

[0348] According to the preparation method of intermediate 1-2, intermediate 37-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 37-2 (85%). ESI-MS: m / z = 278 [M+H] + .

[0349] Step 3: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 3,3-(pyrrolidine-3-yl)phenyl-1,3-diamine (compound 37)

[0350] A white solid (36%) was prepared using intermediates 22-1 and 37-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.29(s,1H),8.79(s,1H),7.96(dt,J=11.0,2.5Hz,1H),7.81(dd,J=8.0,2.0Hz,1 H),7.71(d,J=4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.10–7.01(m,2H),6.9 5(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.19(dd,J=8.0,2.0Hz,1H),5.41(d,J=6.5Hz,1H),3.69(m,1H) ,2.89(m,2H),2.71(m,1H),2.61(dd,J=11.0,4.0Hz,1H),1.91(n,1H),1.53(m,1H); ESI-MS:m / z=389[M+H] + .

[0351] Preparation Example 38: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(piperidin-4-yl)phenyl-1,3-diamine (compound 38)

[0352]

[0353] Step 1: Synthesis of tert-butyl 4-((3-nitrophenyl)amino)piperidine-1-carboxylate (intermediate 38-1)

[0354] A yellow oily substance 38-1 (83%) was prepared from 3-iodonitrobenzene and tert-butyl 4-aminopiperidine-1-carboxylate according to the method in step 1 of Example 1. ESI-MS: m / z = 322 [M+H]+.

[0355] Step 2: Synthesis of tert-butyl 4-((3-aminophenyl)amino)piperidine-1-carboxylate (intermediate 38-2)

[0356] According to the preparation method of intermediate 1-2, intermediate 38-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 38-2 (78%). ESI-MS: m / z = 292 [M+H] + .

[0357] Step 3: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 3,3-(piperidin-4-yl)phenyl-1,3-diamine (compound 38)

[0358] A white solid (57%) was prepared using intermediates 22-1 and 38-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.25(s,1H),8.78(s,1H),8.00(dt,J=11.0,2.0Hz,1H),7.84(dd,J=8.5,2.0Hz,1H),7.73(d,J=4. 0Hz,1H),7.59(dd,J=8.0,7.0Hz,1H),7.20(td,J=8.5,2.6Hz,1H),7.05(dd,J=8.0,2.0Hz,1H),7.00(d,J=2.0Hz,1H),6.94 (t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.21(dd,J=8.0,2.2Hz,1H),5.16(d,J=8.0Hz,1H),3.22–3.16(m,1H),3.15(s,1H ),2.94(dt,J=12.5,3.5Hz,2H),2.46(dd,J=12.5,2.5Hz,2H),1.90–1.83(m,2H),1.27–1.17(m,2H); ESI-MS:m / z=403[M+H] + .

[0359] Preparation Example 39: N 1 -(azacyclobutane-3-yl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 39)

[0360]

[0361] Step 1: Synthesis of tert-butyl 3-((3-nitrophenyl)amino)azacyclobutane-1-carboxylate (intermediate 39-1)

[0362] A yellow oily substance 39-1 (65%) was prepared from 3-iodonitrobenzene and tert-butyl 3-aminoazacyclobutane-1-carboxylate according to the method in step 1 of Example 1. ESI-MS: m / z = 294 [M+H] + .

[0363] Step 2: Synthesis of tert-butyl 3-((3-aminophenyl)amino)azacyclobutane-1-carboxylate (intermediate 39-2)

[0364] Following the preparation method of intermediate 1-2, intermediate 39-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 39-2 (93%). ESI-MS: m / z = 264 [M+H] + .

[0365] Step 3: N 1 -(azacyclobutane-3-yl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 39)

[0366] A white solid (46%) was prepared from intermediates 22-1 and 39-2 according to step 4 of Example 1. ¹H NMR (500 MHz, DMSO-d6) δ 9.30 (d, J = 7.0 Hz, 1H), 8.79 (s, 1H), 7.98 (d, J = 11.0 Hz, 1H), 7.85–7.80 (m, 1H), 7.73 (d, J = 4.0 Hz, 1H), 7.61 (q, J = 8.0 Hz, 1H), 7.22 (td, J = 8.5, 2.5 Hz, 1H), 7.19–7.15 (m, 1H) ),6.97(t,J=8.0Hz,1H),6.88(s,1H),6.69(d,J=4.0Hz,1H),6.14(dd,J=8.0,2.5Hz,1H),5.93(d ,J=6.5Hz,1H),4.12(m,1H),3.69(t,J=7.0Hz,2H),3.40(t,J=7.0Hz,2H); ESI-MS:m / z=375[M+H] + .

[0367] Preparation Example 40: N 1 -(azacyclobutane-3-ylmethyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 40)

[0368]

[0369] Step 1: Synthesis of tert-butyl 3-(((3-nitrophenyl)amino)methyl)azacyclobutane-1-carboxylate (intermediate 40-1)

[0370] A yellow oily substance 40-1 (57%) was prepared from 3-iodonitrobenzene and tert-butyl 3-(aminomethyl)azacyclobutane-1-carboxylate according to the method in step 1 of Example 1. ESI-MS: m / z = 308 [M+H] + .

[0371] Step 2: Synthesis of tert-butyl 3-(((3-aminophenyl)amino)methyl)azacyclobutane-1-carboxylate (intermediate 40-2)

[0372] According to the preparation method of intermediate 1-2, intermediate 40-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 40-2 (93%). ESI-MS: m / z = 278 [M+H] + .

[0373] Step 3: N 1 -(azacyclobutane-3-ylmethyl)-N 3Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 40)

[0374] A white solid (36%) was prepared using intermediates 22-1 and 40-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.28(s,1H),8.79(s,1H),7.97(d,J=11.0Hz,1H),7.82(d,J=8.0Hz ,1H),7.72(d,J=4.0Hz,1H),7.59(q,J=8.0Hz,1H),7.21(t,J=9.0Hz,1H),7.06(d,J=8.0Hz, 1H),7.02(s,1H),6.95(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.20(d,J=8.0Hz,1H),5.44( s,1H),3.78(m,2H),3.28(m,2H),3.12(d,J=6.0Hz,2H),2.84(m,1H); ESI-MS:m / z=389[M+H] + .

[0375] Preparation Example 41: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(pyrrolidine-3-ylmethyl)phenyl-1,3-diamine (compound 41)

[0376]

[0377] Step 1: Synthesis of (S)-3-((3-nitrophenyl)amino)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 41-1)

[0378] A yellow oily substance 41-1 (63%) was prepared from 3-iodonitrobenzene and (S)-3-(aminomethyl)pyrrolidine-1-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 322 [M+H] + .

[0379] Step 2: Synthesis of (S)-3-((3-aminophenyl)amino)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 41-2)

[0380] According to the preparation method of intermediate 1-2, intermediate 41-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 41-2 (86%). ESI-MS: m / z = 292 [M+H] + .

[0381] Step 3: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(pyrrolidine-3-ylmethyl)phenyl-1,3-diamine (compound 41)

[0382] A white solid (55%) was prepared using intermediates 22-1 and 41-2 as raw materials according to step 4 of Example 1. ESI-MS: m / z = 403 [M+H] + .

[0383] Preparation Example 42: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(pyrrolidine-3-ylmethyl)phenyl-1,3-diamine (compound 42)

[0384]

[0385] Step 1: Synthesis of (R)-3-((3-nitrophenyl)amino)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 42-1)

[0386] A yellow oily substance 42-1 (74%) was prepared from 3-iodonitrobenzene and (R)-3-(aminomethyl)pyrrolidine-1-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 322 [M+H] + .

[0387] Step 2: Synthesis of (R)-3-((3-aminophenyl)amino)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (intermediate 42-2)

[0388] According to the preparation method of intermediate 1-2, intermediate 42-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 42-2 (82%). ESI-MS: m / z = 292 [M+H] + .

[0389] Step 3: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(pyrrolidine-3-ylmethyl)phenyl-1,3-diamine (compound 42)

[0390] A white solid (40%) was prepared using intermediates 22-1 and 42-2 as raw materials according to step 4 of Example 1. 1H NMR (500MHz, DMSO-d6) δ9.28(s,1H),8.79(s,1H),7.99(dt,J=11.0,2.5Hz,1H),7.83(dd,J=8.0,2.0Hz,1H),7.73(d,J=4.0H z,1H),7.59(td,J=8.0,6.5Hz,1H),7.20(td,J=8.5,2.5Hz,1H),7.08(d,J=8.0Hz,1H),7.02(t,J=2.5Hz,1H),6.96(t,J=8.0 Hz,1H),6.69(d,J=4.0Hz,1H),6.22(dd,J=8.0,2.0Hz,1H),5.47(s,1H),4.12(s,2H),2.91(d,J=7.0Hz,2H),2.89–2.82(m,1 H),2.82–2.70(m,1H),2.30(dd,J=13.5,7.0Hz,1H),1.98–1.73(m,1H),1.34(dq,J=13.5,7.0Hz,1H).ESI-MS:m / z=403[M+H] + .

[0391] Preparation Example 43: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(morpholino-2-ylmethyl)phenyl-1,3-diamine (compound 43)

[0392] Step 1: Synthesis of (S)-2-(((3-nitrophenyl)amino)methyl)morpholine-4-carboxylic acid tert-butyl ester (intermediate 43-1)

[0393] A yellow oily substance 43-1 (63%) was prepared from 3-iodonitrobenzene and (S)-2-(aminomethyl)morpholine-4-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 338 [M+H] + .

[0394] Step 2: Synthesis of (S)-2-(((3-aminophenyl)amino)methyl)morpholine-4-carboxylic acid tert-butyl ester (intermediate 43-2)

[0395] According to the preparation method of intermediate 1-2, intermediate 43-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 43-2 (71%). ESI-MS: m / z = 308 [M+H] + .

[0396] Step 3: (R)-N 1-(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(morpholino-2-ylmethyl)phenyl-1,3-diamine (compound 43)

[0397]

[0398] A white solid (69%) was prepared using intermediates 22-1 and 43-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.29(s,1H),8.79(s,1H),8.07–7.93(m,1H),7.91–7.80(m,1H),7.73(d,J=4.0Hz,1H),7.60(q,J =8.0Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.09(d,J=8.0Hz,1H),7.06–7.01(m,1H),6.96(t,J=8.0Hz,1H),6.69(d,J=4.0 Hz,1H),6.32–6.17(m,1H),5.29(t,J=6.0Hz,1H),3.74(d,J=11.0Hz,1H),3.51(qd,J=6.5,3.5Hz,1H),3.43(td,J=11.0, 3.5Hz,1H),2.98(t,J=10.0,6.0Hz,2H),2.89–2.77(m,1H),2.70–2.60(m,2H),2.41–2.33(m,1H); ESI-MS: m / z=419[M+H] + .

[0399] Preparation Example 44: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(morpholino-2-ylmethyl)phenyl-1,3-diamine (compound 44)

[0400]

[0401] Step 1: Synthesis of (R)-2-(((3-nitrophenyl)amino)methyl)morpholine-4-carboxylic acid tert-butyl ester (intermediate 44-1)

[0402] A yellow oily substance 44-1 (69%) was prepared from 3-iodonitrobenzene and (R)-2-(aminomethyl)morpholine-4-carboxylic acid tert-butyl ester according to the method in step 1 of Example 1. ESI-MS: m / z = 338 [M+H] + .

[0403] Step 2: Synthesis of (R)-2-(((3-aminophenyl)amino)methyl)morpholine-4-carboxylic acid tert-butyl ester (intermediate 44-2)

[0404] Following the preparation method of intermediate 1-2, intermediate 44-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 44-2 (86%). ESI-MS: m / z = 308 [M+H] + .

[0405] Step 3: (S)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1,3-(morpholino-2-ylmethyl)phenyl-1,3-diamine (compound 44)

[0406] A white solid (53%) was prepared using intermediates 22-1 and 44-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.29(s,1H),8.79(s,1H),7.97(dt,J=11.0,2.5Hz,1H),7.85(dd,J=8.0,2.0Hz,1H),7.73(d,J=4.0Hz,1H),7.60( td,J=8.0,6.0Hz,1H),7.20(td,J=8.0,2.5Hz,1H),7.08(dd,J=8.0,2.0Hz,1H),7.03(t,J=2.0Hz,1H),6.96(t,J=8.0Hz,1H),6.69(d,J=4. 0Hz,1H),6.22(dd,J=8.0,2.0Hz,1H),5.29(t,J=6.0Hz,1H),3.74(dt,J=11.0,2.5Hz,1H),3.51(m,1H),3.43(td,J=11.0,4.0Hz,1H),2.9 6(td,J=6.0,2.0Hz,2H),2.81(dd,J=12.0,2.5Hz,1H),2.64(qd,J=11.0,5.5Hz,2H),2.36(dd,J=12.0,10.0Hz,1H); ESI-MS: m / z=419[M+H] + .

[0407] The structures of compounds 45–48 are as follows:

[0408]

[0409] Preparation Example 45: N 1-(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(1-Methylpiperidin-4-yl)phenyl-1,3-diamine (Compound 45)

[0410] Step 1: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1-(1-methylpiperidin-4-yl)phenyl-1,3-diamine (compound 45)

[0411] Compound 38 (60 mg, 0.15 mmol) and triethylamine (30 mg, 0.30 mmol) were dissolved in anhydrous DCM (2 ml). Iodomethane (26 mg, 0.18 mmol) was added under ice bath conditions, and the mixture was reacted at room temperature for 3 h. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using DCM:NH3 / MeOH (15:1) as the eluent to give a white solid (31 mg, 50%). 1 H NMR(500MHz, CDCl3)δ8.70(s,1H),7.82(dt,J=10.5,2.0Hz,1H),7.53(dt,J=8.0,1.5Hz,1H),7.48(td,J=8 .0,6.0Hz,1H),7.38(t,J=2.0Hz,1H),7.26(s,1H),7.17(s,1H),7.11(t,J=8.0Hz,1H),7.06(td,J=8.0,2. 5Hz,1H),6.74(dd,J=8.0,2.0Hz,1H),6.59(d,J=3.5Hz,1H),6.27(dd,J=8.0,2.5Hz,1H),3.62(s,1H),3.2 8(m,1H),2.83(d,J=11.0Hz,2H),2.32(s,3H),2.16–2.05(m,4H),1.57–1.46(m,2H); ESI-MS:m / z=417[M+H] + .

[0412] Preparation Example 46: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(1-(oxecyclobutan-3-yl)piperidin-4-yl)phenyl-1,3-diamine (compound 46)

[0413] Step 1: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3Synthesis of 1-(1-(oxetane-3-yl)piperidin-4-yl)phenyl-1,3-diamine (compound 46)

[0414] Oxycyclobutane-3-one (25 mg, 0.35 mmol) and sodium cyanoborohydride (33 mg, 0.52 mmol) were dissolved in DMF (2 ml). After stirring at room temperature for 1 h, compound 38 (70 mg, 0.17 mmol) was added, and 2-3 drops of glacial acetic acid were added dropwise to adjust the pH to 5-6. The mixture was stirred overnight at room temperature. The reaction solution was extracted three times with ethyl acetate and water, and washed three times with saturated brine. The organic layer was dried over anhydrous sodium sulfate, and the solvent was recovered under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography using DCM:NH3 / MeOH (25:1) as the eluent to give a white solid (57 mg, 73%). 1 H NMR (500MHz, DMSO-d6) δ9.26(s,1H),8.78(s,1H),8.00(dt,J=11.0,2.5Hz,1H),7.83(dd,J=8.0,2.0Hz,1H),7.73(d,J= 4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.04(dd,J=8.0,2.0Hz,2H),6.94(t,J=8.0Hz,1H), 6.69(d,J=4.0Hz,1H),6.23–6.18(m,1H),5.20(d,J=8.0Hz,1H),4.52(t,J=6.5Hz,2H),4.41(t,J=6.0Hz,2H),3.35(m,1 H),3.11(m,1H),2.68–2.58(m,2H),1.94–1.85(m,2H),1.79(t,J=11.0Hz,2H),1.43–1.31(m,2H); ESI-MS:m / z=459[M+H] + .

[0415] Preparation Example 47: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -(1-(methylsulfonyl)piperidin-4-yl)phenyl-1,3-diamine (compound 47)

[0416] Step 1: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 1-(1-(methylsulfonyl)piperidin-4-yl)phenyl-1,3-diamine (compound 47)

[0417] Compound 38 (60 mg, 0.15 mmol) and triethylamine (30 mg, 0.30 mmol) were dissolved in anhydrous DCM (2 ml). 0.3 ml of a 1 M DCM solution of methanesulfonyl chloride was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was transferred to room temperature and reacted for 1 h. The solvent was recovered by vacuum distillation to obtain the residue, which was then purified by silica gel column chromatography using DCM:NH3 / MeOH (25:1) as the eluent to give a white solid (63 mg, 87%). 1 H NMR(500MHz,DMSO-d6)δ9.28(s,1H),8.79(s,1H),8.00(dt,J=11.0,2.5Hz,1H),7.83(dd,J=8.0,2.0Hz,1H), 7.73(d,J=4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.22(td,J=8.5,2.5Hz,1H),7.09–7.04(m,2H),6.97(t,J =8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.26–6.22(m,1H),5.33(d,J=8.0Hz,1H),3.51(dt,J=11.5,4.0Hz,2H),3 .26(m,1H),2.87(s,3H),2.80(td,J=11.5,2.5Hz,2H),1.98(m,2H),1.49–1.38(m,2H); ESI-MS:m / z=481[M+H] + .

[0418] Preparation Example 48: 1-(4-((3-(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)phenyl)amino)piperidin-1-yl)-2-methylprop-2-ol (Compound 48)

[0419] Step 1: Synthesis of 1-(4-((3-(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)phenyl)amino)piperidin-1-yl)-2-methylprop-2-ol (compound 48)

[0420] Compound 38 (60 mg, 0.15 mmol) and DIPEA (58 mg, 0.45 mmol) were dissolved in THF:MeOH = 1:1 (1 ml: 1 ml). 2,2-Dimethylethylene oxide (32 mg, 0.45 mmol) was added at room temperature, and the mixture was refluxed overnight. The solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using DCM:MeOH (25:1) as the eluent to give a white solid (51 mg, 72%). 1H NMR (500MHz, DMSO-d6) δ9.25(s,1H),8.78(s,1H),8.00(d,J=11.0Hz,1H),7.84(d,J=8.0Hz,1H),7.73(d,J=4.0 Hz,1H),7.59(q,J=8.0Hz,1H),7.20(t,J=8.5Hz,1H),7.10–6.97(m,2H),6.94(t,J=8.0Hz,1H),6.69(d,J=4.0H z,1H),6.20(d,J=8.0Hz,1H),5.14(d,J=8.0Hz,1H),4.02(s,1H),3.06(m,1H),2.89(d,J=11.0Hz,2H),2.18(s, 2H), 2.14 (d, J=12.0Hz, 2H), 1.84 (d, J=12.0Hz, 2H), 1.38 (q, J=11.0Hz, 2H), 1.08 (s, 6H); ESI-MS: m / z=475[M+H] + .

[0421] Preparation Example 49: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -Methyl-N 3 -(piperidin-4-yl)phenyl-1,3-diamine (compound 49)

[0422]

[0423] Step 1: Synthesis of tert-butyl 4-(3-nitrophenylaminomethyl)piperidine-1-carboxylate (intermediate 49-1)

[0424] Intermediate 38-1 (90 mg, 0.28 mmol) was dissolved in DMF (3 mL). NaH (10 mg, 0.42 mmol) was slowly added under ice bath conditions. After stirring at room temperature for 0.5 h, methyl iodide (199 mg, 1.4 mmol) was added, and the reaction was continued at room temperature for another 5 h. After the reaction was complete, the solvent was recovered under reduced pressure to obtain the residue, which was purified by silica gel column chromatography using PE:EA = 8:1 as the eluent to give a yellow oily substance 49-1 (61 mg, 65%). ESI-MS: m / z = 336 [M+H] + .

[0425] Step 2: Synthesis of tert-butyl 4-(3-aminophenylaminomethyl)piperidine-1-carboxylate (intermediate 49-2)

[0426] According to the preparation method of intermediate 1-2, intermediate 49-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 49-2 (86%). ESI-MS: m / z = 306 [M+H] + .

[0427] Step 3: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 -Methyl-N 3 Synthesis of 1,3-(piperidin-4-yl)phenyl-1,3-diamine (compound 49)

[0428] A white solid (63%) was prepared using intermediates 22-1 and 49-2 as raw materials according to step 4 of Example 1. 1 H NMR(500MHz,DMSO-d6)δ9.31(s,1H),8.80(s,1H),7.98(dt,J=11.0,2.5Hz,1 H),7.81(m,1H),7.72(d,J=4.0Hz,1H),7.58(td,J=8.5,6.5Hz,1H),7.28(dd, J=8.0,2.0Hz,1H),7.21(m,1H),7.17(t,J=2.5Hz,1H),7.05(t,J=8.0Hz,1H), 6.69(d,J=4.0Hz,1H),6.41(dd,J=8.5,2.5Hz,1H),3.59(m,1H),3.02-2.96(d J=9.5Hz, 2H), 2.65 (s, 3H), 2.49 (d, J=7.0Hz, 2H), 1.54 (td, J=9.5, 7.0Hz, 4H); ESI-MS: m / z=417[M+H] + .

[0429] Preparation Example 50: N 1 -(3-aminopropyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (Compound 50)

[0430]

[0431] Step 1: Synthesis of tert-butyl (3-((3-nitrophenyl)amino)propyl)carbamate (intermediate 50-1)

[0432] A yellow oily substance 50-1 (31%) was prepared from 3-iodonitrobenzene and tert-butyl (3-aminopropyl)carbamate according to the method in step 1 of Example 1. ESI-MS: m / z = 296 [M+H]+ .

[0433] Step 2: Synthesis of tert-butyl (3-((3-aminophenyl)amino)propyl)carbamate (intermediate 50-2)

[0434] According to the preparation method of intermediate 1-2, intermediate 50-1 was used instead of intermediate 1-1 to obtain transparent oily substance 50-2 (71%). ESI-MS: m / z = 266 [M+H] + .

[0435] Step 3: N 1 -(3-aminopropyl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 50)

[0436] A white solid (43%) was prepared using intermediates 22-1 and 50-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.28 (s, 1H), 8.78 (s, 1H), 7.98 (d, J = 11.0 Hz, 1H), 7.83 (dd, J = 8. 0,2.0Hz,1H),7.72(d,J=4.0Hz,1H),7.59(q,J=8.0Hz,1H),7.21(td,J=8.0,2.5Hz,1H),7 .08(m,1H),7.00(d,J=6.0Hz,1H),6.95(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.19(d, J=8.0Hz,1H),5.34(s,1H),3.00(m,3H),2.64(m,1H),1.63(m,2H); ESI-MS:m / z=377[M+H] + .

[0437] Preparation Example 51: N 1 -(4-aminobutyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 51)

[0438]

[0439] Step 1: Synthesis of tert-butyl (4-((3-nitrophenyl)amino)butyl)carbamate (intermediate 51-1)

[0440] A yellow oily substance 51-1 (57%) was prepared from 3-iodonitrobenzene and tert-butyl (4-aminobutyl)carbamate according to the method in step 1 of Example 1. ESI-MS: m / z = 310 [M+H] + .

[0441] Step 2: Synthesis of tert-butyl (4-((3-aminophenyl)amino)butyl)carbamate (intermediate 51-2)

[0442] According to the preparation method of intermediate 1-2, intermediate 51-1 was used instead of intermediate 1-1 to obtain transparent oily substance 51-2 (85%). ESI-MS: m / z = 280 [M+H] + .

[0443] Step 3: N 1 -(4-aminobutyl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 51)

[0444] A white solid (49%) was prepared using intermediates 22-1 and 51-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.27(s,1H),8.78(s,1H),7.98(dt,J=11.0,2.5Hz,1H),7.82(dd,J=8.0,2.0,1H ),7.72(d,J=4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.06(dd,J=8.0,2.0Hz ,1H),7.00(t,J=2.0Hz,1H),6.94(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.19(dd,J=8.0,2.0Hz,1H),5. 31(s,1H),2.93(t,J=7.0Hz,3H),2.56(t,J=7.0Hz,1H),1.54(m,2H),1.41(m,2H); ESI-MS:m / z=391[M+H] + .

[0445] Preparation Example 52: N 1 -(5-Aminopentyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 52)

[0446]

[0447] Step 1: Synthesis of tert-butyl (5-((3-nitrophenyl)amino)pentyl)carbamate (intermediate 52-1)

[0448] A yellow oily substance 52-1 (72%) was prepared from 3-iodonitrobenzene and tert-butyl (5-aminopentyl)carbamate according to the method in step 1 of Example 1. ESI-MS: m / z = 324 [M+H] + .

[0449] Step 2: Synthesis of tert-butyl (5-((3-aminophenyl)amino)pentyl)carbamate (intermediate 52-2)

[0450] According to the preparation method of intermediate 1-2, intermediate 52-1 was used instead of intermediate 1-1 to obtain transparent oily substance 52-2 (85%). ESI-MS: m / z = 294 [M+H] + .

[0451] Step 3: N 1 -(5-Aminopentyl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 52)

[0452] A white solid (58%) was prepared using intermediates 22-1 and 52-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.26 (s, 1H), 8.78 (s, 1H), 7.98 (dt, J = 11.0, 2.5Hz, 1H), 7.82 (dd, J = 8.0, 2. 0Hz,1H),7.72(d,J=4.0Hz,1H),7.59(td,J=8.0,6.5Hz,1H),7.20(td,J=8.5,2.5Hz,1H),7.07–6.98 (m,2H),6.94(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.18(dd,J=8.0,2.0Hz,1H),5.29(t,J=6.0Hz ,1H),2.92(q,J=6.5Hz,2H),2.53(d,J=6.0Hz,2H),1.51(m,2H),1.34(m,4H); ESI-MS:m / z=405[M+H] + .

[0453] Preparation Example 53: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N3 -(3-(methylamino)propyl)phenyl-1,3-diamine (compound 53)

[0454] Step 1: Synthesis of methyl (3-((3-nitrophenyl)amino)propyl)carbamate tert-butyl ester (intermediate 53-1)

[0455]

[0456] A yellow oily substance 53-1 (71%) was prepared from 3-iodonitrobenzene and tert-butyl (3-aminopropyl)(methyl)carbamate according to the method in step 1 of Example 1. ESI-MS: m / z = 310 [M+H] + .

[0457] Step 2: Synthesis of methyl (3-((3-aminophenyl)amino)propyl)carbamate tert-butyl ester (intermediate 53-1)

[0458] According to the preparation method of intermediate 1-2, intermediate 53-1 was used instead of intermediate 1-1 to obtain transparent oily substance 53-2 (85%). ESI-MS: m / z = 280 [M+H] + .

[0459] Step 3: N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Synthesis of 3-(methylamino)propyl)phenyl-1,3-diamine (compound 53)

[0460] A white solid (42%) was prepared using intermediates 22-1 and 53-2 as raw materials according to step 4 of Example 1. 1 H NMR (500MHz, DMSO-d6) δ9.27(s,1H),8.79(s,1H),7.98(dt,J=11.0,2.5Hz,1H),7.83(dd,J=8.0,2. 0Hz,1H),7.72(d,J=4.0Hz,1H),7.59(q,J=8.0Hz,1H),7.21(td,J=8.5,2.5Hz,1H),7.06(d,J=8.0Hz ,1H),7.01(m,1H),6.95(t,J=8.0Hz,1H),6.69(d,J=4.0Hz,1H),6.19(dd,J=8.0,2.0Hz,1H),5.37(s ,1H),3.78(m,2H),3.08–2.69(m,3H),2.54(s,1H),2.28(m,2H),1.67(m,2H); ESI-MS:m / z=391[M+H] + .

[0461] Preparation Example 54: N 1 -(3-(dimethylamino)propyl)-N 3 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 54)

[0462]

[0463] Step 1: N 1 N 1 -dimethyl-N 3 Synthesis of 1,3-(3-nitrophenyl)propane-1,3-diamine (intermediate 54-1)

[0464] With 3-iodonitrobenzene and N 1 N 1 Using dimethylpropane-1,3-diamine as a starting material, a yellow oily substance 54-1 (62%) was prepared according to the method in step 1 of Example 1. ESI-MS: m / z = 224 [M+H] + .

[0465] Step 2: N 1 N 1 -dimethyl-N 3 Synthesis of 1,3-(3-aminophenyl)propane-1,3-diamine (intermediate 54-2)

[0466] According to the preparation method of intermediate 1-2, intermediate 54-1 was used instead of intermediate 1-1 to obtain transparent oily substance 54-2 (69%). ESI-MS: m / z = 194 [M+H] + .

[0467] Step 3: N 1 -(3-(dimethylamino)propyl)-N 3 Synthesis of 7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)phenyl-1,3-diamine (compound 54)

[0468] Using intermediates 22-1 and 54-2 as raw materials, the mixture was prepared according to step 4 of Example 1 without the need for deprotection of the Boc protecting group, yielding a white solid (17%). 1H NMR(500MHz, CDCl3)δ8.70(s,1H),7.78(d,J=10.5Hz,1H),7.54(d,J=8.0Hz,1H),7.47 (q,J=8.0Hz,1H),7.25(s,1H),7.11(t,J=8.0Hz,1H),7.05(dt,J=8.5,4.0Hz,1H),6.8 4(d,J=8.0Hz,1H),6.58(d,J=4.0Hz,1H),6.29(d,J=8.5Hz,1H),4.14(s,1H),3.15(t, J=7.0Hz, 2H), 2.41 (t, J=7.0Hz, 2H), 2.27 (s, 6H), 1.79 (m, 2H); ESI-MS: m / z=405[M+H] + .

[0469] Preparation Example 55: N 2 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 6 -(piperidin-4-yl)pyridine-2,6-diamine (compound 55)

[0470]

[0471] Step 1: Synthesis of 4-((6-nitropyridin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 55-1)

[0472] A yellow oily substance 55-1 (90%) was prepared from 2-iodo-6-nitropyridine and tert-butyl 4-aminopiperidine-1-carboxylate according to the method in step 1 of Example 1. ESI-MS: m / z = 323 [M+H] + .

[0473] Step 2: Synthesis of tert-butyl 4-((6-aminopyridin-2-yl)amino)piperidine-1-carboxylate (intermediate 55-2)

[0474] Following the preparation method of intermediate 1-2, intermediate 55-1 was used instead of intermediate 1-1 to obtain a pale yellow oily substance 55-2 (83%). ESI-MS: m / z = 293 [M+H] + .

[0475] Step 3: N 2 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 6 Synthesis of -(piperidin-4-yl)pyridine-2,6-diamine (compound 55)

[0476] A white solid (48%) was prepared using intermediates 22-1 and 55-2 as raw materials according to step 4 of Example 1. ESI-MS: m / z = 404 [M+H] + .

[0477] Preparation Examples 56-69: (R)-N 1 -(7-(3-fluorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-N 3 Salt formation compounds of -(piperidin-3-yl)phenyl-1,3-diamine

[0478]

[0479] acid = citric acid, hemifumaric acid, malic acid, L-malic acid, D-malic acid, methanesulfonic acid, L-tartaric acid, D-tartaric acid, succinic acid, maleic acid, formic acid, acetic acid, hydrochloric acid, phosphoric acid

[0480] Compound 32 was dissolved in isopropanol, and a solution containing 1.2 equivalents of organic or inorganic acid was slowly added dropwise at room temperature. After the addition was complete, the reaction mixture was stirred overnight at 40°C. The reaction mixture was cooled to room temperature, filtered, and the solid was washed with diethyl ether and dried to obtain the corresponding salts (56, citrate of compound 32; 57, hemifumarate of compound 32; 58, malate of compound 32; 59, L-malate of compound 32; 60, D-malate of compound 32; 61, methanesulfonate of compound 32; 62, L-tartrate of compound 32; 63, D-tartrate of compound 32; 64, succinate of compound 32; 65, maleate of compound 32; 66, formate of compound 32; 67, acetate of compound 32; 68, hydrochloride of compound 32; 69, phosphate of compound 32).

[0481] Bioactivity testing section

[0482] Kinase activity test of the compounds of this invention

[0483] The inhibitory activity of the compounds provided in this invention against FLT3, FLT3-D835Y and IRAK4 kinases was tested.

[0484] Instrument: Envision microplate reader TM (PerkinElmer, USA).

[0485] Materials: Human recombinant FLT3, purchased from Carna Biosciences, FLT3 protein fused with GST (aa564-59993), FLT3 activity assay kit KinEASETMTK, purchased from Cisbio; Human recombinant FLT3-D835Y, purchased from Eurofins, FLT3 protein fragment fused with GST (containing D835Y mutation) (aa564-end), FLT3-D835Y activity assay kit KinEASETMTK, purchased from Cisbio; Human recombinant IRAK4, purchased from SinoBiological.

[0486] Sample preparation: Dissolve DMSO, store at low temperature, and control the concentration of DMSO in the final system within a range that does not affect the detection activity.

[0487] Experimental procedure: FLT3, FLT3-D835Y and the substrate (specifically biotin-labeled peptide TK Substrate) were diluted with HTRF kinase buffer (1.25X kinase buffer, 6.25mM MgCl2, 1.25mM MnCl2, 1.25mM MDT), respectively. IRAK4 and the substrate were diluted with HTRF kinase buffer (1X kinase buffer, 5mM MgCl2, 1mM MnCl2, 1mM MDT). 4 μL of enzyme, 4 μL of substrate, and 2 μL of different concentrations of the test compounds were added to a 384 reaction plate (ProxiPlate™-384Plus, PerkinElmer). The specific reaction systems were: FLT3: 2% DMSO, 0.5 ng / μL FLT3, 1 μM TK-S, 2 μM ATP; FLT3-D835Y: 2% DMSO, 0.4 ng / μL FLT3-D835Y, 1 μM TK-S, 1 μM ATP; IRAK4: 2% DMSO, 2 ng / μL IRAK4, 1 μM TK-S, 100 μM ATP. After incubation at room temperature for 1 hour, antibodies were added for detection. A control group (using DMSO instead of the test compound) and a blank control group were also included. Each sample and each concentration was tested in triplicate. Initial screening was performed at a single concentration, for example, 1 μmol, to test the activity of the samples. For samples that exhibit activity under certain conditions, such as an inhibition rate (%inhibition) greater than 50%, the activity dose-dependent relationship, i.e., IC50, is tested. 50The value was obtained by nonlinearly fitting the sample concentration to the sample activity. The calculation software used was Graphpad Prism 8, and the fitting model used was sigmoidal dose-response (variable slope). For most inhibitor screening models, the bottom and top of the fitting curve were set to 0 and 100, respectively.

[0488] Table 1. Inhibitory activities of the compounds of the present invention against FLT3, FLT3-D835Y, and IRAK4 kinases.

[0489]

[0490]

[0491] IC50: Half-inhibitory concentration

[0492] A: IC 50 <20nM;B:20nM <IC 50 <200nM; C:200nM <IC 50 <500nM; D:

[0493] 500nM <IC 50 <5000nM

[0494] CA4948:

[0495] The results in Table 1 show that most of the compounds exhibited good FLT3 and IRAK4 kinase inhibitory activity, indicating that these compounds have the potential to treat FLT3, IRAK4, and FLT3 / IRAK4 related diseases.

[0496] Cell proliferation inhibitory activity test of the compounds of this invention

[0497] Cell lines: MV4-11 (human acute myeloid monocytic leukemia, expressing FLT3-ITD homozygous mutation), Molm-13 (human acute myeloid monocytic leukemia, expressing FLT3-ITD heterozygous mutation), TF-1 (human hematologic leukemia cell, expressing IRAK4).

[0498] Experimental procedure: The antiproliferative activity (IC50) of the test compound against MV-4-11 and other cell lines was determined by MTS assay. 50Cells in logarithmic growth phase were digested with trypsin, counted, and seeded at a density of 1×10⁴ cells / well in 96-well plates (100 cells per well). The plates were incubated overnight at 37°C with 5% CO₂. Six concentration gradients were set up for each test compound, with three replicates for each concentration. After addition, the plates were incubated for 72 h, and then 20 μL MTS was added. After incubation at 37°C for 2 h, the absorbance at 490 nm (L1) was measured using a SpectraMAX 340 microplate reader, with a reference wavelength of 690 nm (L2). The (L1-L2) values ​​were plotted against different inhibitor concentrations, and the IC50 of the compounds was calculated using GraphPad Prism 5 software. 50 (Inhibition rate = (OD value of control group - OD value of treatment group) / OD value of control group * 100%)

[0499] Table 2. Inhibitory activities of the compounds of the present invention against MV4-11, MOLM13, and TF-1.

[0500]

[0501] IC50: Half-inhibitory concentration

[0502] A: IC 50 <10nM; B:10nM <IC 50 <100nM; C:100nM <IC 50 <500nM; D:500nM <IC 50 <5000nM

[0503] Table 2 above shows that the compounds of the present invention have good inhibitory activity against the proliferation of MV4-11, MOLM13 and TF-1 cells.

Claims

1. A compound, characterized in that, It has the structure shown in general formula (I): Or its optical isomer or its pharmaceutically acceptable salt; Wherein, ring A is selected from benzene ring or 5-6 membered heteroaromatic ring, wherein the heteroatom in the heteroaromatic ring is 1 to 4 N atoms, and the benzene ring or heteroaromatic ring may be further divided by 1 to 3 R atoms. 4 replace; X is selected from CH or N; Y is selected from N and CR. X ;R X Selected from H, deuterium, halogens, C 1-3 alkyl, C 1-3 Alkyloxy groups, halogenated C 1-3 Alkyl, CN, NO2; R 1 Selected from H, deuterium, halogens, C 1-3 Alkyl, halogen-substituted C 1-3 Alkyl, C 1-3 alkyloxy groups, -NR a R b 、-C(=O)-NR a R b 、-S(=O)2-NR a R b ; R 2 R 3 Each is independently selected from H, deuterium, halogen, and C. 1-3 Alkyl, halogen-substituted C 1-3 Alkyl, C 1-3 Alkyloxy group, CN, NO2, -C(=O)-NR aa R bb -S(=O)2-NR aa R bb -L 1 -NR aa R bb -L 1 -R 5 ; L 1 It is a chemical bond, -O(CH2) o -、-NR 7 (CH2) p -; R a R b R aa R bb Each is independently selected from H, deuterium, OH, and C. 1-3 alkyl, C 3-5 cycloalkyl, -(C=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups); o and p are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8; R 4 Selected from H, C 1-5 alkyl, C 3-8 Cycloalkyl groups, 4- to 8-membered heterocyclic alkyl groups, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 Alkyl groups, phenyl groups, and 5-8 heteroaryl groups, wherein the alkyl, cycloalkyl, phenyl, and heteroaryl groups can be independently separated by 0, 1, or 2 R groups. 8 Replaced by multiple R 8 When replacing, multiple R 8 They can be independent and may be the same or different; R 5 Selected from H, OH, CN, halogens, C 1-3 alkyl, C 1-3 Halogenated alkyl, C 4-8 The alkyl group, cycloalkyl group, or heterocycloalkyl group of 4 to 8 members, wherein the alkyl group, cycloalkyl group, or heterocycloalkyl group may be further substituted with 0, 1, or 2 R groups. 6 replace; R 6 Selected from H, NH2, -NH-C 1-3 Alkyl, -N-(C 1-3 alkyl)2, C 1-5 alkyl, C 3-8 Cycloalkyl groups, 4- to 8-membered heterocyclic alkyl groups, -C(=O)-(C 1-3 Alkyl groups), -CHO, -S(=O)2-(C 1-3 The alkyl group, wherein the cycloalkyl group may be further substituted with 0, 1 or 2 hydroxyl groups; R 7 For H, C 1-3 Alkyl groups; R 8 For H, deuterium, halogen, OH, NH2, CF3, OCH3, CN, NO2, C 1-5 Alkyl group, -C(=O)NH-L 2 -R 9 The alkyl group may be further substituted with 0, 1, or 2 hydroxyl groups or amino groups; L 2 For chemical bonds, C 1-8 alkylene s and t are independently selected from 1, 2, and 3, respectively; R 9 The components are H, OH, and NH2.

2. The compound according to claim 1, characterized in that, A is selected from: in Represents the substituent junction; the R 4 The definition is the same as in claim 1.

3. The compound according to claim 2, characterized in that, It has the structure shown in general formula (II): The R 1 R 2 R 3 R 4 The definition is the same as in claim 1.

4. The compound according to claim 1 or 3, characterized in that, The R 1 H, -NR a R b ;R 2 For -NH(CH2) p -NR aa R bb -NR 7 (CH2) p -R 5 The R a R b R aa R bb Each is independently selected from H, deuterium, methyl, and ethyl; p is selected from 0, 1, 2, 3, 4, and 5; R 3 H, deuterium, halogens; R 4 For H, C 1-5 alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups, -S(=O)2-(C 1-3 Alkyl groups, phenyl groups, and 5-8 heteroaryl groups, wherein the alkyl, cycloalkyl, phenyl, and heteroaryl groups can be independently separated by 0, 1, or 2 R groups. 8 Replace; R 5 Heterocyclic alkyl groups of 4 to 8 members, C 4-8 cycloalkyl; R 6 For H, NH2, methylamino, dimethylamino, methanesulfonyl, C 1-4 Alkyl, 4- to 6-membered heterocyclic alkyl, wherein the alkyl group may be further substituted with 0 or 1 hydroxyl group; R 7 For H, CH3; R 8 For H, deuterium, halogen, OH, CF3, OCH3, C 1-5 The alkyl group may be further substituted with 0 or 1 hydroxyl group.

5. The compound according to claim 1, characterized in that, The compound has the following structure: Where R 2 For -NH(CH2) p -NR aa R bb , R aa R bb Each is independently selected from H, deuterium, and CH3; m, n, and p are independently selected from 1, 2, 3, 4, and 5; R 6 H, methanesulfonyl, C 1-4 The alkyl group or 3- to 6-membered heterocyclic alkyl group may be further substituted with hydroxyl groups.

6. The compound according to any one of claims 1, 3, and 5, characterized in that, A is selected from: R 4 Selected from: H, ethyl, methyl, propyl, cyclopropyl, isopropyl, cyclohexyl, oxetyl, hydroxytert-butyl, methanesulfonyl, phenyl, pyridyl, methylimidazolyl, phenyl with one or two halogen substituted, methylphenyl, methoxyphenyl, trifluoromethylphenyl, hydroxyisopropyl-substituted phenyl, carbamoylphenyl, hydroxy and methyl-substituted butylcarbamoylphenyl; R 1 Selected from H, deuterium, halogens, and ethylamino; Y is selected from N and CH; R 3 Selected from: H, F; R 2 Selected from:

7. The compound according to claim 1, characterized in that, Selected from the following compounds: And optical isomers or pharmaceutically acceptable salts.

8. The compound of claim 1, characterized in that, Pharmaceutically acceptable salts include acid addition salts formed by compounds of general formula (I) with the following acids: formic acid, acetic acid, propionic acid, pyruvic acid, glycolic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, mandelic acid, citric acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, L-malic acid, D-malic acid, lactic acid, camphor sulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, salicylic acid, benzoic acid, tartaric acid, L-tartaric acid, D-tartaric acid, oxalic acid, succinic acid, maleic acid, ascorbic acid, amino acids, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, hydroiodic acid, or perchloric acid.

9. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-8, alone or in combination with other formulations, in the preparation of an antitumor drug or an immune disease drug; wherein the other therapeutic agent is selected from one or more of IDH1 inhibitors, IDH2 inhibitors, Bcl-2 inhibitors, hypomethylating agents, and antimetabolites.

10. The application according to claim 9, characterized in that, The tumors mentioned include bladder cancer, breast cancer, colon cancer, kidney cancer, epidermal cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, or skin cancer; lymphatic spectrum hematopoietic tumors such as acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, pilocellular lymphoma, Burkett's lymphoma, bone marrow spectrum hematopoietic tumors, acute and chronic myeloid leukemia, acute and chronic myeloid leukemia, spinal dysplasia syndrome, promyeloid leukemia, thyroid follicular carcinoma, stromal tumors, fibrosarcoma, rhabdomyosarcoma, central or peripheral nervous system tumors, astrocytoma, neuroblastoma, glioma, schwannoma, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, xanthokeratoma, and Kaposi's sarcoma; The immune diseases mentioned are selected from arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Auder's thyroiditis, Graves' disease, rheumatoid arthritis syndrome, multiple sclerosis, infectious neuritis, acute infectious encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis, graft-versus-host disease, transplantation, transfusion allergy, allergic reaction, type I hypersensitivity reaction, allergic conjunctivitis, allergic rhinitis, and atopic dermatitis.

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