Cyclic compound as well as pharmaceutical composition and application thereof

Abstract

The invention provides a cyclic compound with a structure as shown in a formula (I) or a pharmaceutically acceptable salt or a stereoisomer or a prodrug molecule of the cyclic compound, and a pharmaceutical composition and application of the cyclic compound or the pharmaceutically acceptable salt or the stereoisomer or the prodrug molecule of the cyclic compound. The compound can be used as a protein kinase inhibitor, has very strong inhibitory activity on FLT3 mutant kinase, has very strong proliferation inhibitory activity on Ba / F3-FLT3-ITD and stable strain cells containing drug-resistant point mutation, can inhibit proliferation, migration and invasion of various tumor cells, and especially can overcome the drug resistance of existing clinical drugs. The cyclic compound provided by the invention can be used for preparing medicines for preventing or treating FLT3 tyrosine kinase mediated diseases (such as hematological malignancies), and can be used for treating various hematological malignancies of human beings and other mammals. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the technical field of chemical medicine, and particularly relates to a cyclic compound, its pharmaceutical composition and application. Background Art

[0002] FLT3 (Fms-like tyrosine kinase) belongs to the members of the type III receptor tyrosine kinase family, which is mainly expressed on the cell surface of normal hematopoietic stem cells and hematopoietic progenitor cells, and its ligand is mainly expressed in bone marrow stromal cells. FLT3 ligand can selectively stimulate the proliferation of CD34+ progenitor cells, specifically bind to FLT3, induce the receptor to dimerize and autophosphorylate tyrosine residues in the kinase domain, further activate downstream signaling pathways, mainly including RAS / RAF / MEK, PI3K / AKT and JAK / STAT5, etc., and participate in regulating cell proliferation, differentiation and cell survival. Studies have found that FLT3 mutations lead to constitutive abnormal activation of FLT3, resulting in abnormal proliferation of tumor cells such as hematopoietic cells and lymphocytes, and causing a variety of malignant blood diseases.

[0003] FLT3 mutations mainly include two types: internal tandem duplication mutations (FLT3-ITD) in the juxtamembrane domain and point mutations (FLT3-TKD) in the tyrosine kinase domain. The first-generation marketed FLT3 inhibitors such as midostaurin lack specificity for FLT3 and often cause toxicity and adverse reactions due to off-target effects, limiting the clinical treatment effect. The second-generation FLT3 selective inhibitors gilteritinib and quizartinib were approved by the FDA for marketing in 2018 and 2023 respectively. They can overcome various mutations of FLT3, including FLT3-ITD mutations and various FLT3-TKD mutations (D835Y / V / N / H / E, Y842C / H, etc.), but have poor inhibitory effects on the gatekeeper residue F691L mutation, resulting in a significant decline in the clinical treatment effect of the drug. Currently, there is no inhibitor for the F691L mutation on the market. Summary of the Invention

[0004] Based on this, the purpose of the present invention is to provide a novel inhibitor against FLT3 mutations.

[0005] In order to achieve the above purpose, the present invention includes the following technical solutions.

[0006] On the one hand, the present invention provides a cyclic compound having the structure shown in formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof:

[0007]

[0008]

[0009] Among them, E, Z, and Q are each independently selected from: CR 1 or N;

[0010] Each R 1 is independently selected from: H, halogen, R 2 substituted or unsubstituted C1-C6 alkyl, R 2 substituted or unsubstituted C1-C6 alkoxy, R 2 substituted or unsubstituted C3-C8 cycloalkyl;

[0011] Each R 2 is independently selected from: H, halogen, C1-C6 alkyl, C1-C6 alkoxy;

[0012] X, Y, K, and D are each independently selected from: -O-, -C(R 3 R 3 )-, -N(R 3 )C(R 3 R 3 )-, -N(R 3 )-, -S-, -S(=O)-, -S(O)2-, -C(=O)-, -(C=O)N(R 3 )-;

[0013] Each R 3 is independently selected from: H, C1-C6 alkyl;

[0014] L is selected from: R 4 substituted or unsubstituted C2-C 12 alkylene, R 4 substituted or unsubstituted C2-C 12 oxaalkylene, R 4 substituted or unsubstituted C2-C 12 thiaalkylene, R 4 substituted or unsubstituted C2-C 12 unsaturated hydrocarbon group;

[0015] Each R 4 is independently selected from: H, halogen, C1-C6 alkyl;

[0016] A is selected from: R 5 substituted or unsubstituted C3-C6 cycloalkyl, R 5 substituted or unsubstituted 4-7 membered saturated nitrogen-containing heterocyclic group;

[0017] Each R 5 is independently selected from: H, halogen, nitro, hydroxy, cyano, C1-C6 alkyl, C1-C6 alkoxy;

[0018] B is selected from: H, halogen, nitro, hydroxy, cyano, R6 Substituted or unsubstituted C1-C 20 alkyl, R 6 Substituted or unsubstituted C1-C 20 alkoxy, R 6 Substituted or unsubstituted C3-C 12 cycloalkyl, R 6 Substituted or unsubstituted 3- to 12-membered heterocyclic group, R 6 Substituted or unsubstituted 5- to 10-membered heteroaryl, -NR 7 R 8 ;

[0019] Each R 6 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy, R 9 Substituted or unsubstituted 3- to 8-membered heterocyclic group, -NR 3 R 3 ;

[0020] R 7 、R 8 are independently selected from: H, R 6 Substituted or unsubstituted C1-C6 alkyl, R 6 Substituted or unsubstituted 3- to 12-membered heterocyclic group, or R 7 、R 8 together with the nitrogen atom to which it is attached forms R 6 Substituted or unsubstituted 3- to 12-membered heterocyclic group;

[0021] Each R 9 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy.

[0022] In some embodiments, the cyclic compound has the structure shown in formula (II):

[0023]

[0024] In some embodiments, R 1 is selected from: H, C1-C6 alkyl; preferably methyl, ethyl, n-propyl, isopropyl.

[0025] In some embodiments, X, Y, K, D are independently selected from: -O-, -N(R 3 )-, -N(R 3 )C(R 3 R 3 )-, -(C=O)N(R 3 )-, -C(=O)-, -C(R 3 R3 )-; R 3 Selected from: H, C1-C3 alkyl.

[0026] In some embodiments, both X and Y are -N(H)-.

[0027] In some embodiments, K is selected from: -O-, -(C=O)N(H)-.

[0028] In some embodiments, D is selected from: -C(=O)-, -CH2-, -N(H)-.

[0029] In some embodiments, L is selected from: R 4 Substituted or unsubstituted C3-C8 alkylene, -(CR 2 R 2 ) n -O-(CR 2 R 2 ) m -, -(CR 2 R 2 ) n -S-(CR 2 R 2 ) m -, R 4 Substituted or unsubstituted C3-C8 unsaturated hydrocarbon group;

[0030] Wherein, n and m are each independently selected from positive integers between 1 and 7, and n + m is not greater than 8;

[0031] Each R 2 Is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl;

[0032] Each R 4 Is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl.

[0033] In some embodiments, L is selected from: -(CH2) a -, -(CH2) n -O-(CH2) m -, -(CH2) b -CH(R 4 )-(CH2) c -;

[0034] Wherein, a is selected from: 3, 4, 5, 6, 7, 8;

[0035] n and m are each independently selected from: 1, 2, 3;

[0036] b and c are each independently selected from: 1, 2, 3;

[0037] R 4 Selected from: methyl, ethyl.

[0038] In some embodiments, L is selected from: -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)2-O-(CH2)2-, -(CH2)2-CH(CH3)-(CH2)2-.

[0039] In some embodiments, A is selected from: a 5- or 6-membered saturated nitrogen-containing heterocyclic group, and the nitrogen atom is connected to D.

[0040] In some embodiments, A is selected from:

[0041] In some embodiments, B is selected from: H, R 6 Substituted or unsubstituted C1-C6 alkyl, R 6 Substituted or unsubstituted C1-C6 alkoxy, R 6 Substituted or unsubstituted C3-C8 cycloalkyl, R 6 Substituted or unsubstituted 3- to 8-membered heterocyclic group, R 6 Substituted or unsubstituted 5- to 10-membered heteroaryl, -NR 7 R 8 ;

[0042] Each R 6 Is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C3 alkyl, C1-C3 alkoxy, R 9 Substituted or unsubstituted 4- to 6-membered heterocyclic group, -NR 3 R 3 ;

[0043] R 7 、R 8 Are independently selected from: H, R 6 Substituted or unsubstituted C1-C3 alkyl, R 6 Substituted or unsubstituted 3- to 8-membered heterocyclic group, or R 7 、R 8 Together with the nitrogen atom to which it is attached forms R 6 Substituted or unsubstituted 3- to 8-membered heterocyclic group;

[0044] Each R 9 Is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C3 alkyl, C1-C3 alkoxy;

[0045] Each R 3 Is independently selected from: H, C1-C3 alkyl.

[0046] In some of these embodiments, B is selected from: H, -NR 7 R 8 ;

[0047] R 7 、R 8 are each independently selected from: H, R 6 -substituted or unsubstituted C1-C3 alkyl, R 6 -substituted or unsubstituted 5- to 6-membered heterocyclic group, or R 7 、R 8 together with the nitrogen atom to which it is attached form a R 6 -substituted or unsubstituted 5- to 6-membered heterocyclic group;

[0048] Each R 6 is independently selected from: H, hydroxy, methyl, ethyl, propyl, methoxy, ethoxy, R 9 -substituted or unsubstituted 4- to 6-membered heterocyclic group, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, dipropylamino;

[0049] Each R 9 is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl, methoxy, ethoxy.

[0050] In some of these embodiments, B is selected from: H, methyl, ethyl, methoxy, ethoxy, cyclopentyl, cyclohexyl,

[0051]

[0052] Each R 6 is independently selected from: H, methyl, ethyl, propyl, dimethylamino, methanesulfonyl;

[0053] R 9 is selected from: H, methyl, ethyl, propyl, dimethylamino, methanesulfonyl.

[0054] In some of these embodiments, X and Y are both -N(H)-, K is -O-, and D is -CH2-;

[0055] L is selected from: -(CH2)4-, -(CH2)2-O-(CH2)2-, -(CH2)2-CH(CH3)-(CH2)2-;

[0056] A is

[0057] B is selected from: -NR 7 R 8 ;

[0058] R 7 、R 8independently selected from: methyl, ethyl, propyl, methyl substituted with dimethylamino, ethyl substituted with dimethylamino, propyl substituted with dimethylamino, or R 7 and R 8 together with the nitrogen atom to which it is attached form R 6 a substituted or unsubstituted 5- or 6-membered heterocyclic group;

[0059] each R 6 is independently selected from: H, methyl, ethyl, propyl, R 9 a substituted or unsubstituted 4- to 6-membered heterocyclic group, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, dipropylamino;

[0060] each R 9 is independently selected from: H, methyl, ethyl, propyl.

[0061] On the other hand, the present invention also provides the use of the cyclic compound or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof in the preparation of an FLT3 inhibitor.

[0062] In some embodiments, the FLT3 is a mutated FLT3.

[0063] In some embodiments, the gene mutation is an FLT3-ITD mutation, an FLT3-TKD mutation.

[0064] In some embodiments, the gene mutation is an FLT3 ITD mutation, an FLT3 D835Y mutation, an FLT3 D835Y / F691L mutation, an FLT3 ITD / D835Y mutation, an FLT3 ITD / F691L mutation.

[0065] In a third aspect, the present invention also provides the use of the cyclic compound or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof in the preparation of a drug for preventing and / or treating an FLT3-mediated disease.

[0066] In some embodiments, the FLT3-mediated disease is a malignant blood disease.

[0067] In some embodiments, the malignant blood disease is acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and myeloproliferative disorder (MPD), etc.

[0068] Fourthly, the present invention also provides a pharmaceutical composition for preventing and / or treating malignant blood diseases, which is prepared from an active ingredient and a pharmaceutically acceptable excipient. The active ingredient includes the cyclic compound described in the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof.

[0069] The present invention provides a novel cyclic compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. Such compounds can be used as protein kinase inhibitors, have strong inhibitory activity against FLT3 mutant kinases, and have strong inhibitory activity against the proliferation of stable cell lines of Ba / F3-FLT3-ITD with drug-resistant point mutations. They can inhibit the proliferation, migration and invasion of various tumor cells, and in particular can overcome the drug resistance of existing clinical drugs. The cyclic compounds provided by the present invention can be used to prepare drugs for preventing or treating diseases mediated by FLT3 tyrosine kinase (such as malignant blood diseases), and can be used to treat various malignant blood diseases of humans and other mammals, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and myeloproliferative disorders (MPD), etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1 It shows the in vivo anti-cancer research results of compound ZX31-9 in the Ba / F3-FLT3-ITD-D835Y xenograft model. DETAILED DESCRIPTION OF THE INVENTION

[0071] The technical solutions of the present invention will be further described below through specific examples. Those skilled in the art should understand that the examples are only for helping to understand the present invention and should not be regarded as specific limitations to the present invention.

[0072] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not used to limit the present invention.

[0073] The terms "comprising" and "having" and any variations thereof in the present invention are intended to cover non-exclusive inclusion. For example, a process, method, device, product or equipment that includes a series of steps is not limited to the listed steps or modules, but optionally further includes steps not listed, or optionally further includes other steps inherent to these processes, methods, products or equipment.

[0074] As used herein, "plural" means two or more. "And / or" describes the relationship between related objects and indicates that three relationships may exist. For example, A and / or B may mean: A exists alone, A and B exist simultaneously, or B exists alone. The character " / " generally indicates that the related objects before and after are in an "or" relationship.

[0075] In the compounds of the present invention, when any variable (e.g., R 4 , R 5 etc.) appears more than once in any component, its definition for each occurrence is independent of the definition for each other occurrence. Similarly, combinations of substituents and variables are allowed, provided that such combinations render the compound stable. The line extending a substituent into a ring system indicates that the indicated bond may be attached to any ring atom capable of substitution. If the ring system is polycyclic, it means that such a bond is attached only to any suitable carbon atom of an adjacent ring. It is to be understood that one of ordinary skill in the art may select the substituents and substitution patterns of the compounds of the present invention to provide compounds that are chemically stable and can be readily synthesized from readily available starting materials by techniques known in the art and the methods set forth below. If a substituent itself is substituted by more than one group, it is to be understood that these groups may be on the same carbon atom or on different carbon atoms, provided that the structure is stable.

[0076] As used herein, the term "alkyl" means a branched and straight-chain saturated aliphatic hydrocarbon group having a specific number of carbon atoms. For example, the definition of "C1-C6 alkyl" for "C1-C6" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight-chain or branched-chain manner. For example, "C1-C6 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0077] As used herein, the term "alkylene" means a residue obtained by removing two hydrogen atoms from a branched and straight-chain aliphatic hydrocarbon having a specific number of carbon atoms, i.e., a group having one less hydrogen atom than "alkyl". For example, "C1-C6 alkylene" specifically includes -CH2- (methylene), -(CH2)2- (ethylene), -(CH2)3- (propylene), -(CH2)4- (butylene), -(CH2)5- (pentylene), -(CH2)6- (hexylene), etc.

[0078] As used herein, the term "unsaturated hydrocarbon group" means a branched and straight-chain unsaturated aliphatic hydrocarbon group having a specific number of carbon atoms, i.e., a non-cyclic unsaturated chain hydrocarbon group, and the carbon chain contains one or more carbon-carbon double bonds or contains a carbon-carbon triple bond, such as: -CH=CHCH2-, -(CH2)8(CH=CH)CH2-, -(CH2)7CH=CH-, -(CH2)8CH=CH-, etc.

[0079] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic, bicyclic or polycyclic hydrocarbon group in which the ring atoms are composed of carbon atoms. The bicyclic or polycyclic groups include spiro rings, fused rings and bridged rings. For example, "cycloalkyl" includes, but is not limited to, the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0080] As used herein, the term "alkoxy" refers to "-O-alkyl", such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0081] As used herein, the term "oxaalkylene" refers to a group composed of "alkylene - O - alkylene", such as -CH2OCH2-, -CH2OCH2CH2-, -CH2CH2OCH2CH2-, -CH2CH2OCH2CH2 - OCH2CH2-, etc.

[0082] As used herein, the term "thiaalkylene" refers to a group composed of "alkylene - S - alkylene", such as -CH2SCH2-, -CH2SCH2CH2-, -CH2CH2SCH2CH2-, -CH2CH2SCH2CH2 - OCH2CH2-, etc.

[0083] As used herein, the term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic or polycyclic ring substituent in which one or more ring atoms are heteroatoms selected from N, O or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. The bicyclic or polycyclic groups include spiro rings, fused rings and bridged rings. For example: morpholinyl, piperidinyl, pyrrolidinyl, pyrrolidinyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, tetrahydrofuryl, tetrahydrothienyl, etc., and their N - oxides. The connection of the heterocyclic substituent can be achieved through a carbon atom or through a heteroatom.

[0084] As used herein, the term "heteroaryl" refers to an aromatic ring containing one or more heteroatoms selected from O, N, or S, which aromatic ring may be monocyclic, bicyclic, or polycyclic, and includes, for example, but is not limited to: quinolinyl, pyrazolyl, pyrrolyl, thienyl, furyl, pyridyl, pyrimidinyl, pyrazinyl, triazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridazinyl, 1,3,5-triazinyl, benzofuranyl, benzothienyl, indolyl, indazolyl, benzimidazolyl, benzopyrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzopyrimidinyl, pyridopyrimidinyl, pyridopyridyl, purinyl, pteridinyl, imidazothiazolyl, imidazopyridazinyl, pyrazolopyrimidinyl, etc.; "heteroaryl" is also understood to include any N-oxide derivative of a heteroaryl containing nitrogen. The heteroaryl can be linked through a carbon atom or through a heteroatom.

[0085] As will be understood by those skilled in the art, "halo" or "halogen" as used in the present invention means chlorine, fluorine, bromine, and iodine.

[0086] In one embodiment of the present invention, the present invention provides a cyclic compound having the structure shown in formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof:

[0087]

[0088] Wherein, E, Z, and Q are each independently selected from: CR 1 or N;

[0089] Each R 1 is independently selected from: H, halogen, R 2 substituted or unsubstituted C1-C6 alkyl, R 2 substituted or unsubstituted C1-C6 alkoxy, R 2 substituted or unsubstituted C3-C8 cycloalkyl;

[0090] Each R 2 is independently selected from: H, halogen, C1-C6 alkyl, C1-C6 alkoxy;

[0091] X, Y, K, and D are each independently selected from: -O-, -C(R 3 R 3 )-, -N(R 3 )C(R 3 R 3 )-, -N(R 3 )-, -S-, -S(=O)-, -S(O)2-, -C(=O)-, -(C=O)N(R 3 );

[0092] Each R 3Independently selected from: H, C1-C6 alkyl;

[0093] L is selected from: R 4 Substituted or unsubstituted C2-C 12 Alkylene, R 4 Substituted or unsubstituted C2-C 12 Oxoalkylene, R 4 Substituted or unsubstituted C2-C 12 Thiaalkylene, R 4 Substituted or unsubstituted C2-C 12 Unsaturated hydrocarbon group;

[0094] Each R 4 Independently selected from: H, halogen, C1-C6 alkyl;

[0095] A is selected from: R 5 Substituted or unsubstituted C3-C6 cycloalkyl, R 5 Substituted or unsubstituted 4-7 membered saturated nitrogen-containing heterocyclic group;

[0096] Each R 5 Independently selected from: H, halogen, nitro, hydroxy, cyano, C1-C6 alkyl, C1-C6 alkoxy;

[0097] B is selected from: H, halogen, nitro, hydroxy, cyano, R 6 Substituted or unsubstituted C1-C 20 Alkyl, R 6 Substituted or unsubstituted C1-C 20 Alkoxy, R 6 Substituted or unsubstituted C3-C 12 Cycloalkyl, R 6 Substituted or unsubstituted 3-12 membered heterocyclic group, R 6 Substituted or unsubstituted 5-10 membered heteroaryl, -NR 7 R 8 ;

[0098] Each R 6 Independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy, R 9 Substituted or unsubstituted 3-8 membered heterocyclic group, -NR 3 R 3 ;

[0099] R 7 , R 8 Independently selected from: H, R 6 Substituted or unsubstituted C1-C6 alkyl, R 6 Substituted or unsubstituted 3-12 membered heterocyclic group, or R7 , R 8 together with the nitrogen atom to which it is attached forms R 6 a substituted or unsubstituted 3- to 12-membered heterocyclic group;

[0100] Each R 9 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy.

[0101] The present invention includes the free form of the compound of formula (I), as well as its pharmaceutically acceptable salts and stereoisomers. The term "free form" refers to an amine compound in non-salt form. The pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of the present invention containing a basic moiety or an acidic moiety by conventional chemical methods. Generally, salts of basic compounds are prepared by ion exchange chromatography or by reacting the free base with a stoichiometric or excess amount of the desired salt form of an inorganic or organic acid in a suitable solvent or a combination of solvents. Similarly, salts of acidic compounds are formed by reacting with a suitable inorganic or organic base.

[0102] Accordingly, the pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts of the compounds of the present invention formed by reacting the basic compounds of the present invention with inorganic or organic acids. For example, the conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc., and also include salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxy-benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, 2-hydroxyethanesulfonic acid, trifluoroacetic acid, etc.

[0103] If the compound of the present invention is acidic, then the appropriate "pharmaceutically acceptable salts" refer to salts prepared by reacting with pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. Ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases, said bases including salts of primary amines, secondary amines and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, aminoglucose, histidine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0104] Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci.’ 1977:66:1 - 19 describes in more detail the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts.

[0105] In one embodiment, the present invention provides a drug for treating diseases such as malignant blood diseases of humans or other mammals by using the compound having the structure shown in formula (I) and its pharmaceutically acceptable salts, which can be used to treat various malignant blood diseases of humans and other mammals, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and myeloproliferative disorders (MPD), etc.

[0106] Metabolites of the compounds and their pharmaceutically acceptable salts involved in the present invention, as well as prodrugs that can be transformed in vivo into the structures of the compounds and their pharmaceutically acceptable salts involved in the present application, are also included in the claims of the present invention.

[0107] The present invention also provides a pharmaceutical composition, which contains an active ingredient within a safe and effective amount range, as well as a pharmaceutically acceptable carrier or excipient.

[0108] The “active ingredient” described in the present invention refers to the compound of formula I described in the present invention, or its pharmaceutically acceptable salt, or its stereoisomer, or its prodrug molecule.

[0109] The “active ingredient” and the pharmaceutical composition described in the present invention can be used as inhibitors of FLT3 mutant kinases and can be used to prepare drugs for preventing and / or treating malignant blood diseases, etc.

[0110] “Safe and effective amount” means that the amount of the active ingredient is sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 - 2000 mg of the active ingredient per dose, more preferably, it contains 10 - 200 mg of the active ingredient per dose. Preferably, the “per dose” is one tablet.

[0111] “Pharmaceutically acceptable carrier or excipient” means one or more compatible solid or liquid fillers or gelling substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0112] “Compatible” herein means that the components in the composition can be mixed with the active ingredient of the present invention and with each other without significantly reducing the efficacy of the active ingredient.

[0113] Some examples of pharmaceutically acceptable carriers or excipients include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0114] In another preferred embodiment, the compound of formula I of the present invention can form a complex with a macromolecular compound or polymer through non-bonding interactions. In another preferred embodiment, the compound of formula I of the present invention, as a small molecule, can also be linked to a macromolecular compound or polymer through chemical bonds. The macromolecular compound can be a biological macromolecule such as polysaccharide, protein, nucleic acid, polypeptide, etc.

[0115] There is no particular limitation on the administration mode of the active ingredient or pharmaceutical composition of the present invention. Representative administration modes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0116] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

[0117] In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or calcium phosphate, or mixed with the following components:

[0118] (a) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol and silicic acid;

[0119] (b) binders, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0120] (c) humectants, such as glycerol;

[0121] (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

[0122] (e) slow solvents, such as paraffin wax;

[0123] (f) absorption accelerators, such as quaternary ammonium compounds;

[0124] (g) wetting agents, such as cetyl alcohol and glycerol monostearate;

[0125] (h) adsorbents, such as kaolin; and

[0126] (i) Lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0127] The solid dosage forms can also be prepared with coatings and shell materials such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active ingredient in such compositions can be delayed and released in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and wax substances.

[0128] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 - butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and fragrances.

[0129] In addition to the active ingredient, the suspension may contain suspending agents such as ethoxylated isooctadecanol, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum monostearate, and agar, or mixtures of these substances.

[0130] Compositions for parenteral injection may contain physiologically acceptable sterile aqueous or non - aqueous solutions, dispersions, suspensions, or emulsions, and sterile powders for re - dissolving into sterile injectable solutions or dispersions. Suitable aqueous and non - aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0131] The compounds of the present invention can be administered alone or in combination with other therapeutic drugs.

[0132] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage administered is a pharmaceutically effective dosage. For a person weighing 60 kg, the daily dosage is generally 1 - 2000 mg, preferably 20 - 500 mg. Of course, the specific dosage should also consider factors such as the route of administration and the health status of the patient, which are within the scope of the skills of a skilled physician.

[0133] The compounds of formula I can be used in combination with other drugs known for treating or ameliorating similar conditions. When administered in combination, the mode of administration and dosage of the original drug remain unchanged, while the compound of formula I is taken simultaneously or subsequently. When the compound of formula I is taken simultaneously with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the compound of formula I is preferably used. The combination of drugs also includes taking the compound of formula I and one or more known drugs during overlapping time periods. When the compound of formula I is used in combination with one or more other drugs, the dosage of the compound of formula I or the known drug may be lower than the dosage when they are used alone.

[0134] The present invention will be further illustrated with reference to specific examples. It should be understood that these examples are only for illustrating the present invention and not for limiting the scope of the present invention. The experimental methods without specific conditions noted in the following examples are generally carried out under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are by weight.

[0135] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the methods of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0136] The raw materials in the following examples can be obtained commercially, or prepared by methods known in the art, or prepared according to the methods described herein.

[0137] Example 1: Preparation of 11-oxo-6-oxa-2,4-diaza-3(2,4)-pyrimidin-1(4,1)-piperidin-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX8-8)

[0138]

[0139] Step 1: Preparation of ethyl 5-(3-nitrophenoxy)valerate (Compound 2)

[0140]

[0141] Dissolve compound 1 (3 g, 14.3 mmol) in 50 mL of anhydrous DMF in a 200 mL round-bottom flask. Add NaH (0.69 g, 28.7 mmol) in three portions at 0 °C. After stirring for 0.5 h, add ethyl 5-bromovalerate (3.6 g, 17.3 mmol) to the mixture. After reacting at room temperature for 6 h, slowly add the reaction solution to ice water and extract with EA and water 2-3 times. After drying the organic layer with anhydrous Na2SO4 and rotary evaporation, column chromatography gives 4.26 g of white solid (yield: 89%).

[0142] 1 1H NMR (400 MHz, DMSO-d6) δ 7.10 (t, J = 8.0 Hz, 1H), 6.58 - 6.41 (m, 3H), 4.12 (q, J = 7.1 Hz, 2H), 3.91 (d, J = 5.4 Hz, 2H), 2.37 (d, J = 6.8 Hz, 2H), 1.79 (d, J = 5.7 Hz, 4H), 1.47 (s, 9H), 1.25 (d, J = 7.1 Hz, 3H). LC-MS (ESI) m / z 336.0 [M-H] - .

[0143] Step 2: Preparation of ethyl 5-(3-aminophenoxy)valerate (Compound 3)

[0144]

[0145] Dissolve compound 2 (4.26 g, 12.65 mmol) in 40 ml of DCM. After adding 10 ml of TFA, react at room temperature for 3 h. Extract with DCM and water 2-3 times. After drying the organic layer with anhydrous Na2SO4 and rotary evaporation of the solvent, 2.75 g of crude brown oil (crude yield: 92%) is obtained and directly used for the next step of the reaction.

[0146] Step 3: Preparation of ethyl 5-(3-((2-chloro-5-cyanopyrimidin-4-yl)amino)phenoxy)valerate (Compound 4)

[0147]

[0148] Dissolve compound 3 (2.75 g, 11.63 mmol) in 50 mL of 1,4-dioxane. Add 2,4-dichloropyrimidine-5-carbonitrile (2.01 g, 11.63 mmol) and DIEA (3.84 mL, 23.26 mmol). After reacting at 50 °C for 8 h, rotary evaporate the reaction system and obtain 2.31 g of white solid by column chromatography (yield: 51%).

[0149] 11H NMR (400 MHz, DMSO-d6) δ 8.41 (s, 1H), 7.96 (s, 1H), 7.24 (d, J = 8.3 Hz, 1H), 7.20 - 7.17 (m, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.70 (dd, J = 8.3, 2.4 Hz, 1H), 3.96 - 3.90 (m, 2H), 3.80 (d, J = 6.8 Hz, 2H), 2.39 (d, J = 7.1 Hz, 2H), 1.79 (s, 4H), 1.22 (d, J = 7.1 Hz, 3H). LC-MS (ESI) m / z 389.2 [M-H] - .

[0150] Step 4: Preparation of tert-butyl 4-((5-cyano-4-((3-((5-ethoxy-5-oxopentyl)oxy)phenyl)amino)pyrimidin-2-yl)amino)piperidine-1-carboxylate (Compound 5)

[0151]

[0152] Dissolve Compound 4 (2.31 g, 5.93 mmol) in 50 ml of 1,4-dioxane, add tert-butyl 4-aminopiperidine-1-carboxylate (1.42 g, 7.11 mmol) and DIEA (1.96 mL, 11.86 mmol). After reacting at 50 °C for 8 hours, rotary evaporate the reaction system, and obtain 1.44 g of white solid by column chromatography (yield: 45%).

[0153] 1 1H NMR (400 MHz, DMSO-d6) δ 9.53 (s, 1H), 8.54 (s, 1H), 8.12 (d, J = 4.4 Hz, 1H), 7.85 - 7.82 (m, 2H), 7.45 (t, J = 9.1 Hz, 1H), 6.75 (d, J = 5.6 Hz, 1H), 4.11 - 4.06 (m, 3H), 4.01 (t, J = 6.4 Hz, 2H), 3.60 - 3.51 (m, 2H), 2.85 (s, 2H), 2.47 (t, J = 7.4 Hz, 2H), 2.36 - 2.20 (m, 2H), 2.02 - 1.81 (m, 6H), 1.35 (s, 9H), 1.21 (t, J = 7.1 Hz, 3H). LC-MS (ESI) m / z 537.0 [M-H] - .

[0154] Step 5: Preparation of 5-(3-((2-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-5-cyanopyrimidin-4-yl)amino)phenoxy)pentanoic acid (Compound 6)

[0155]

[0156] Compound 5 (1.44 g, 2.67 mmol) was dissolved in a mixed solvent of 12 mL of THF / H2O = 5:1. After adding LiOH (0.32 g, 13.35 mmol), the mixture was stirred overnight at 50 °C. The reaction system was evaporated to dryness, and 0.68 g of white solid (yield 50%) was obtained by column chromatography and directly used for the next step.

[0157] Step 6: Preparation of 11-oxo-6-oxa-2,4-diazaperimidine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 -carbonitrile (Compound 7)

[0158]

[0159] Compound 6 (0.68 g, 1.33 mmol) was dissolved in 12 mL of DCM. 3 mL of TFA was added, and the mixture was reacted at room temperature for 2 hours. Then it was extracted 3 times with ethyl acetate and saturated NaHCO3 solution. The organic layer was dried over anhydrous Na2SO4 and evaporated to dryness to obtain a brownish oil. The crude product was dissolved in 300 mL of DMF, and DIPEA (2.19 mL, 13.3 mmol) and FDPP (0.77 g, 1.99 mmol) were added. After reacting for 6 h, it was extracted 3 times with DCM and H2O. The organic layer was dried over anhydrous Na2SO4 again and evaporated to dryness. 0.13 g of white solid was obtained by column chromatography (total yield of two steps: 25%).

[0160] 1 H NMR (400 MHz, DMSO-d6) δ 12.31 (s, 1H), 9.76 (s, 1H), 8.75 (d, J = 4.7 Hz, 1H), 7.54 (s, 1H), 7.30 (t, J = 8.1 Hz, 1H), 6.76 (d, J = 8.3 Hz, 1H), 6.64 (d, J = 7.9 Hz, 1H), 3.93 - 3.84 (m, 2H), 3.61 - 3.53 (m, 3H), 2.79 (s, 2H), 2.37 - 2.21 (m, 2H), 2.18 - 1.98 (m, 2H), 1.93 - 1.69 (m, 6H). LC-MS (ESI) m / z 391.3 [M-H] - .

[0161] Step 7: Preparation of 11-oxo-6-oxa-2,4-diazaperimidine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 -carboxamide (Compound ZX8-8)

[0162]

[0163] Dissolve compound 7 (0.13 g, 0.25 mmol) in 5 mL of DMSO. Sequentially add 3 mL of NaOH solution (2 mol / L) and 2 mL of 30% aqueous H2O2 solution at 0 °C. After reacting at room temperature for 10 minutes, extract with EA and H2O three times. After drying the organic layer with anhydrous Na2SO4 and rotary evaporation, column chromatography gives 30 mg of white solid (yield: 22%).

[0164] 1 H NMR (400 MHz, DMSO-d6) δ 11.76 (s, 1H), 8.53 (s, 1H), 7.88 - 7.74 (m, 3H), 7.20 (t, J = 8.2 Hz, 2H), 6.59 (d, J = 8.1 Hz, 1H), 6.48 (d, J = 8.0 Hz, 1H), 4.02 - 3.94 (m, 3H), 3.76 - 3.73 (m, 1H), 3.43 (s, 1H), 3.24 - 3.18 (m, 1H), 3.05 (s, 1H), 2.46 - 2.38 (m, 2H), 2.23 - 2.09 (m, 2H), 1.88 - 1.66 (m, 6H). LC-MS (ESI) m / z 409.3 [M-H] - .

[0165] Example 2: 5 4 -morpholino-11-oxo-6-oxo-2,4-diaza-3(2,4)-pyrimidine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 Preparation of -carboxamide (Compound ZX9-9)

[0166]

[0167] The synthesis method is as in Example 1.

[0168] 11H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 8.51 (s, 1H), 7.83 (s, 1H), 7.75 (s, 1H), 7.57 (s, 1H), 7.19 (s, 1H), 6.80 (d, J = 8.4 Hz, 1H), 6.48 (d, J = 8.3 Hz, 1H), 3.91 - 3.79 (m, 2H), 3.72 (s, 4H), 3.63 - 3.49 (m, 4H), 3.17 (d, J = 4.9 Hz, 1H), 3.03 - 2.89 (m, 4H), 2.43 - 2.26 (m, 2H), 2.05 - 1.99 (m, 2H), 1.85 - 1.81 (m, 6H). LC-MS (ESI) m / z 493.9 [M-H] - .

[0169] Example 3: 5 4 -((2-Methoxyethyl)(methyl)amino)-11-oxo-6-oxa-2,4-diaza-3(2,4)-pyrimidine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX13-9) Preparation

[0170]

[0171] The synthesis method was as in Example 1.

[0172] 1 1H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 8.51 (s, 1H), 7.83 (s, 1H), 7.73 (s, 1H), 7.55 (s, 1H), 7.18 (s, 1H), 6.80 (d, J = 8.2 Hz, 1H), 6.45 (d, J = 8.2 Hz, 1H), 3.88 - 3.76 (m, 2H), 3.69 - 3.62 (m, 2H), 3.50 (t, J = 5.2 Hz, 2H), 3.33 (s, 3H), 3.23 (s, 3H), 3.17 (t, J = 5.8 Hz, 2H), 2.74 (s, 3H), 2.43 - 2.25 (m, 2H), 2.01 (s, 2H), 1.87 - 1.76 (m, 6H). LC-MS (ESI) m / z 496.1 [M-H] - .

[0173] Example 4: 3 3 -Ethyl-11-oxo-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX16-7) Preparation

[0174]

[0175] Step 1: Preparation of tert-butyl 4-((6-bromo-5-cyano-3-ethylpyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 2)

[0176]

[0177] Dissolve Compound 1 (1.0 g, 4.08 mmol) in 10 mL of 1,4-dioxane, and successively add DIPEA (1.35 mL, 8.16 mmol) and tert-butyl 4-aminopiperidine-1-carboxylate (0.98 g, 4.90 mmol). After reacting at 50 °C for 8 hours, evaporate the reaction system to dryness, and obtain 1.25 g of white solid by column chromatography (yield: 75%).

[0178] 1 H NMR (400 MHz, DMSO-d6) δ 7.62 (d, J = 7.8 Hz, 1H), 4.16 - 4.04 (m, 1H), 3.97 - 3.94 (m, 2H), 2.84 (s, 2H), 2.61 (q, J = 7.3 Hz, 2H), 1.79 (dd, J = 12.6, 3.9 Hz, 2H), 1.53 - 1.44 (m, 2H), 1.41 (s, 9H), 1.16 (d, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 408.1 [M-H] - .

[0179] Step 2: Preparation of tert-butyl 4-((5-cyano-6-(3-((5-ethoxy-5-oxopentyl)oxy)phenyl)amino)-3-ethylpyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 3)

[0180]

[0181] Add Compound 2 (1.24 g, 2.93 mmol), ethyl 5-(3-aminophenoxy)valerate (0.6 g, 2.53 mmol), palladium acetate (56 mg, 0.25 mmol), Xantphos (0.3 g, 0.51 mmol), cesium carbonate (1.65 g, 5.06 mmol) and 50 mL of toluene to a 100 mL two-necked flask. Replace the reaction system with argon three times, seal the reaction system, stir at 60 °C for 3 hours, filter the reaction solution through diatomaceous earth, evaporate the solvent to obtain a black mixture, and obtain 0.86 g of yellow solid by column chromatography (yield: 50%).

[0182] 11H NMR (400 MHz, Chloroform-d) δ 7.31 (t, J = 2.3 Hz, 1H), 7.22 (t, J = 8.1 Hz, 1H), 7.03 (d, J = 8.1 Hz, 1H), 6.83 (s, 1H), 6.62 (dd, J = 8.2, 2.4 Hz, 1H), 4.87 (d, J = 7.1 Hz, 1H), 4.17 - 4.07 (m, 5H), 4.01 - 3.98 (m, 2H), 3.96 - 3.92 (m, 2H), 2.54 (q, J = 7.5 Hz, 2H), 2.41 - 2.37 (m, 2H), 2.13 - 2.09 (m, 2H), 1.85 - 1.82 (m, 4H), 1.49 - 1.44 (m, 11H), 1.30 (s, 3H), 1.28 (d, J = 2.5 Hz, 3H). LC-MS (ESI) m / z 565.1 [M-H] - .

[0183] Step 3: Preparation of 5-(3-((6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-3-cyano-5-ethylpyrazin-2-yl)amino)phenoxy)pentanoic acid (Compound 4)

[0184]

[0185] Compound 4 was obtained by referring to the synthesis method of Step 5 in Example 1, and the crude product was directly used in the next step.

[0186] Step 4: 3 3 -Ethyl-11-oxo-6-oxo-2,4-diaza-3(2,6)-pyrazino-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 -carbonitrile (Compound 5)

[0187]

[0188] Compound 5 was obtained by referring to the synthesis method of Step 6 in Example 1.

[0189] 11H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 7.34 (t, J = 2.2 Hz, 1H), 7.18 - 7.13 (m, 2H), 6.82 (d, J = 8.0 Hz, 1H), 6.55 (dd, J = 8.1, 2.4 Hz, 1H), 4.06 - 4.01 (m, 2H), 3.97 - 3.91 (m, 1H), 3.80 - 3.72 (m, 2H), 3.21 - 3.15 (m, 2H), 2.60 - 2.53 (m, 2H), 2.45 - 2.34 (m, 2H), 2.23 - 2.13 (m, 4H), 1.86 - 1.82 (m, 4H), 1.13 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 419.0 [M-H] - .

[0190] Step 5: 3 3 -Ethyl-11-oxo-6-oxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX16-7) Preparation

[0191]

[0192] Dissolve Compound 5 (83 mg, 0.20 mmol) in 5 mL of DMSO. Add 3 mL of NaOH solution (2 mol / L) and 1.5 mL of 30% aqueous H2O2 solution successively at 0 °C. After reacting at room temperature for 10 minutes, extract with EA and H2O three times. The organic layer is dried over anhydrous Na2SO4 and then concentrated by rotary evaporation. Column chromatography gives 29 mg of white solid (yield: 25%).

[0193] 1 1H NMR (400 MHz, DMSO-d6) δ 11.40 (s, 1H), 7.78 (s, 1H), 7.58 (d, J = 3.2 Hz, 1H), 7.27 (d, J = 3.2 Hz, 1H), 7.18 - 7.14 (m, 2H), 6.50 (dd, J = 8.2, 2.4 Hz, 1H), 6.43 (dd, J = 8.0, 2.1 Hz, 1H), 3.97 (d, J = 4.3 Hz, 2H), 3.88 (s, 1H), 3.75 - 3.72 (m, 2H), 3.30 - 3.16 (m, 2H), 2.60 - 2.54 (m, 2H), 2.45 - 2.38 (m, 2H), 2.26 - 2.15 (m, 2H), 1.96 - 1.70 (m, 6H), 1.20 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 437.1 [M-H] -.

[0194] Example 5: 3 3 -Methyl-11-oxo-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -Carboxamide (Compound ZX39-10) Preparation

[0195]

[0196] The synthesis method is as in Example 4.

[0197] 1 H NMR (400 MHz, DMSO-d6) δ 11.42 (s, 1H), 7.77 (s, 1H), 7.65 (s, 1H), 7.24 (s, 1H), 7.17 - 7.11 (m, 2H), 6.48 (d, J = 8.2 Hz, 1H), 6.42 (d, J = 8.0 Hz, 1H), 3.96 - 3.88 (m, 3H), 3.72 - 3.66 (m, 2H), 3.31 - 3.17 (m, 2H), 2.42 (s, 2H), 2.25 (s, 3H), 2.20 - 2.16 (m, 2H), 1.91 - 1.71 (m, 6H). LC-MS (ESI) m / z 423.1 [M-H] - .

[0198] Example 6: 3 3 -Isopropyl-11-oxo-6-oxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -Carboxamide (Compound ZX40-10) Preparation

[0199]

[0200] The synthesis method is as in Example 4.

[0201] 11H NMR (400 MHz, DMSO-d6) δ 11.36 (s, 1H), 7.78 (s, 1H), 7.54 (s, 1H), 7.27 (s, 1H), 7.20 (d, J = 7.1 Hz, 1H), 7.16 (t, J = 8.1 Hz, 1H), 6.50 (dd, J = 8.2, 2.4 Hz, 1H), 6.43 (d, J = 8.0 Hz, 1H), 3.99 - 3.95 (m, 2H), 3.80 - 3.69 (m, 3H), 3.24 (s, 2H), 3.14 - 3.08 (m, 1H), 2.47 - 2.39 (m, 2H), 2.24 - 2.14 (m, 2H), 1.99 - 1.72 (m, 6H), 1.16 (dd, J = 6.7, 3.3 Hz, 6H). LC-MS (ESI) m / z 451.3 [M-H] - .

[0202] Example 7: 3 3 -Ethyl-5 4 -(4-Methylpiperazin-1-yl)-11-oxo-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX17-8) Preparation

[0203]

[0204] The synthesis method was as in Example 4.

[0205] 1 1H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 7.68 (s, 1H), 7.53 (s, 1H), 7.19 (s, 1H), 6.94 (d, J = 7.2 Hz, 1H), 6.79 (d, J = 8.4 Hz, 1H), 6.42 (dd, J = 8.4, 2.1 Hz, 1H), 3.94 - 3.89 (m, 2H), 3.79 - 3.74 (m, 1H), 3.68 - 3.49 (m, 6H), 3.27 (s, 2H), 2.97 (s, 4H), 2.60 - 2.54 (m, 2H), 2.27 (s, 3H), 2.03 - 1.85 (m, 10H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 535.3 [M-H] - .

[0206] Example 8: 3 3 -Ethyl-5 4-Morpholino-11-oxo-6-oxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 Preparation of -carboxamide (Compound ZX18-8)

[0207]

[0208] The synthesis method is as in Example 4.

[0209] 1 H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.69 (s, 1H), 7.53 (s, 1H), 7.20 (s, 1H), 6.95 (d, J = 7.1 Hz, 1H), 6.79 (d, J = 8.4 Hz, 1H), 6.44 (dd, J = 8.4, 2.3 Hz, 1H), 3.95 - 3.89 (m, 2H), 3.81 - 3.75 (m, 1H), 3.72 (t, J = 4.7 Hz, 4H), 3.61 - 3.46 (m, 2H), 3.00 - 2.93 (m, 4H), 2.60 - 2.53 (m, 4H), 2.20 - 2.03 (m, 2H), 1.90 - 1.79 (m, 8H), 1.19 (d, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 522.1 [M-H] - .

[0210] Example 9: 5 4 -((2-(Dimethylamino)ethyl)(methyl)amino)-3 3 -Ethyl-11-oxo-6-oxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 Preparation of -carboxamide (Compound ZX20-8)

[0211]

[0212] The synthesis method is as in Example 4.

[0213] 11H NMR (400 MHz, DMSO-d6) δ 11.17 (s, 1H), 7.71 (s, 1H), 7.54 (s, 1H), 7.22 (s, 1H), 6.92 (d, J = 7.2 Hz, 1H), 6.80 (d, J = 8.4 Hz, 1H), 6.40 (dd, J = 8.4, 2.3 Hz, 1H), 4.00 - 3.88 (m, 2H), 3.76 - 3.73 (m, 1H), 3.60 (s, 2H), 3.49 (s, 2H), 3.04 (d, J = 6.2 Hz, 2H), 2.69 (s, 3H), 2.61 - 2.55 (m, 2H), 2.37 (t, J = 7.2 Hz, 2H), 2.13 (s, 6H), 2.09 - 2.01 (m, 2H), 1.96 - 1.87 (m, 8H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 537.3 [M-H] - .

[0214] Example 10: 3 3 -Ethyl-5 4 -(4-Morpholinopiperidin-1-yl)-11-oxo-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX21-8) Preparation

[0215]

[0216] The synthesis method is as in Example 4.

[0217] 1 1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 7.69 (s, 1H), 7.53 (d, J = 2.7 Hz, 1H), 7.21 (d, J = 2.8 Hz, 1H), 6.93 (d, J = 7.3 Hz, 1H), 6.79 (d, J = 8.4 Hz, 1H), 6.41 (dd, J = 8.4, 2.2 Hz, 1H), 3.95 - 3.88 (m, 2H), 3.78 - 3.76 (m, 1H), 3.66 - 3.57 (m, 6H), 3.43 - 3.36 (m, 2H), 3.32 (s, 8H), 2.58 (q, J = 7.4 Hz, 2H), 2.45 - 2.42 (m, 1H), 2.23 - 2.18 (m, 2H), 2.05 - 1.84 (m, 10H), 1.56 - 1.48 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 605.0 [M-H] - .

[0218] Example 11: 3 3 -Ethyl-5 4 -(4-(Oxetan-3-yl)piperazin-1-yl)-11-oxo-6-oxa-2,4-diazatricyclo[2.6.2.02,7]dodec-3-ene-5 5 -carboxamide (Compound ZX22-8) Preparation

[0219]

[0220] The synthesis method is as in Example 4.

[0221] 1 1 - .

[0222] Example 12: 3 3 -Ethyl-5 4 -(4-(4-Methylpiperazin-1-yl)piperidin-1-yl)-11-oxo-6-oxa-2,4-diazatricyclo[2.6.2.02,7]dodec-3-ene-5 5 -carboxamide (Compound ZX23-8) Preparation

[0223]

[0224] The synthesis method is as in Example 4.

[0225] 11H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.67 (s, 1H), 7.53 (d, J = 2.7 Hz, 1H), 7.21 (d, J = 2.0 Hz, 1H), 6.97 (d, J = 7.2 Hz, 1H), 6.78 (d, J = 8.5 Hz, 1H), 6.41 (dd, J = 8.4, 1.9 Hz, 1H), 4.03 (q, J = 7.1 Hz, 2H), 3.94 - 3.89 (m, 1H), 3.77 - 3.61 (m, 4H), 3.28 (s, 4H), 2.77 (s, 6H), 2.60 - 2.55 (m, 2H), 2.45 (s, 4H), 2.24 - 2.15 (m, 1H), 2.08 - 2.01 (m, 2H), 1.99 (s, 3H), 1.91 - 1.79 (m, 8H), 1.62 - 1.57 (m, 2H), 1.19 (t, J = 3.7 Hz, 3H). LC-MS (ESI) m / z 618.3 [M-H] - .

[0226] Example 13: 3 3 -Ethyl-5 4 -Morpholinyl-6-oxo-2,4,7-triaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX25-9) Preparation

[0227]

[0228] The synthesis method was as in Example 4.

[0229] 11H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.83 (t, J = 5.7 Hz, 1H), 8.30 (d, J = 2.6 Hz, 1H), 7.57 (d, J = 3.3 Hz, 1H), 7.27 (d, J = 3.0 Hz, 1H), 7.19 (d, J = 7.9 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 7.01 (dd, J = 8.6, 2.7 Hz, 1H), 3.92 (s, 1H), 3.72 (t, J = 4.5 Hz, 4H), 3.14 (s, 2H), 3.01 (s, 2H), 2.89 (s, 2H), 2.85 (t, J = 4.5 Hz, 4H), 2.57 (q, J = 7.5 Hz, 2H), 2.03 - 1.97 (m, 2H), 1.92 - 1.89 (m, 2H), 1.83 - 1.76 (m, 6H), 1.17 (d, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 521.1 [M-H] - .

[0230] Example 14: 3 3 -Ethyl-5 4 -Morpholinyl-6-oxa-2,4-diaza-3(2,6)-pyrazin-1(4,1)-piperidin-5(1,3)-benzoncycloundecane-3 5 -Carboxamide (Compound ZX27-8) Preparation

[0231]

[0232] Step 1: Preparation of 2-(Benzyloxy)-1-fluoro-4-nitrobenzene (Compound 2)

[0233]

[0234] Dissolve Compound 1 (1 g, 6.3 mmol) in 40 mL of DMF, and successively add potassium carbonate (1.74 g, 12.6 mmol) and benzyl bromide (0.89 mL, 7.56 mmol). Stir at 70 °C for 3 hours, spin-dry the reaction system, and obtain 1.32 g of yellow solid by column chromatography (yield: 85%).

[0235] 11H NMR (400 MHz, Chloroform-d) δ 7.95 (dd, J = 7.2, 2.7 Hz, 1H), 7.87 - 7.81 (m, 1H), 7.51 - 7.47 (m, 2H), 7.46 - 7.42 (m, 2H), 7.41 - 7.36 (m, 1H), 7.27 - 7.21 (m, 1H), 5.24 (s, 2H). LC-MS (ESI) m / z 246.0 [M-H] - .

[0236] Step 2: Preparation of 4-(2-(Benzyloxy)-4-nitrophenyl)morpholine (Compound 3)

[0237]

[0238] Dissolve Compound 2 (1.32 g, 5.3 mmol) in 40 mL of DMF, and successively add potassium carbonate (1.47 g, 10.6 mmol) and morpholine (0.55 g, 6.36 mmol). Stir at 70 °C for 8 hours, rotary evaporate the reaction system, and obtain 1.25 g of yellow solid by column chromatography (yield: 75%).

[0239] 1 1H NMR (400 MHz, Chloroform-d) δ 7.91 (dd, J = 8.8, 2.5 Hz, 1H), 7.85 (d, J = 2.5 Hz, 1H), 7.49 - 7.42 (m, 4H), 7.41 - 7.36 (m, 1H), 6.92 (d, J = 8.8 Hz, 1H), 5.20 (s, 2H), 3.90 - 3.81 (m, 4H), 3.32 - 3.26 (m, 4H). LC-MS (ESI) m / z 313.1 [M-H] - .

[0240] Step 3: Preparation of 3-(Benzyloxy)-4-morpholinoaniline (Compound 4)

[0241]

[0242] Dissolve Compound 3 (1.25 g, 3.98 mmol) in a mixed solvent of 100 mL of ethanol:water at a ratio of 5:1, add solid NH4Cl (0.85 g, 15.92 mmol) and iron powder (1.11 g, 19.9 mmol), heat to 70 °C and react for 6 hours. After filtering with diatomaceous earth, rotary evaporate the filtrate, and obtain 0.79 g of brown solid by column chromatography (crude yield: 70%). The crude product is directly used for the next step.

[0243] Step 4: Preparation of tert-butyl 4-((6-((3-(benzyloxy)-4-morpholinophenyl)amino)-5-cyano-3-ethylpyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 5)

[0244]

[0245] Compound 5 was obtained by referring to the synthesis method in Step 2 of Example 4.

[0246] 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 1H), 7.47 (d, J = 7.0 Hz, 2H), 7.41 (t, J = 7.6 Hz, 2H), 7.33 (t, J = 7.2 Hz, 1H), 7.26 (d, J = 2.3 Hz, 1H), 7.22 (dd, J = 8.6, 2.3 Hz, 1H), 7.03 (d, J = 7.4 Hz, 1H), 6.86 (d, J = 8.6 Hz, 1H), 5.07 (s, 2H), 3.94 - 3.89 (m, 3H), 3.71 (t, J = 4.5 Hz, 4H), 2.97 (t, J = 4.5 Hz, 4H), 2.64 (s, 2H), 2.56 (t, J = 7.4 Hz, 2H), 1.83 - 1.80 (m, 2H), 1.45 - 1.42 (m, 2H), 1.38 (s, 9H), 1.13 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 612.3 [M-H] - .

[0247] Step 5: Preparation of tert-butyl 4-((6-((3-(benzyloxy)-4-morpholinophenyl)amino)-5-carbamoyl-3-ethylpyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 6)

[0248]

[0249] Compound 6 was obtained by referring to the synthesis method in Step 5 of Example 4.

[0250] 11H NMR (400 MHz, DMSO-d6) δ 11.05 (s, 1H), 7.54 (s, 1H), 7.48 (d, J = 7.1 Hz, 2H), 7.41 (t, J = 7.5 Hz, 2H), 7.37 - 7.31 (m, 2H), 7.23 (s, 1H), 7.17 (d, J = 2.3 Hz, 1H), 6.90 (d, J = 8.6 Hz, 1H), 6.75 (d, J = 7.5 Hz, 1H), 5.14 (s, 2H), 4.10 - 4.02 (m, 1H), 4.00 - 3.89 (m, 2H), 3.75 - 3.67 (m, 4H), 2.97 (t, J = 4.6 Hz, 4H), 2.78 (s, 2H), 2.60 - 2.55 (m, 2H), 1.91 - 1.88 (m, 2H), 1.51 - 1.44 (m, 2H), 1.40 (s, 9H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 630.3 [M-H] - .

[0251] Step 6: Preparation of tert-butyl 4-((5-carbamoyl-3-ethyl-6-(3-hydroxy-4-morpholinophenyl)amino)pyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 7)

[0252]

[0253] Dissolve Compound 6 (0.65 g, 1.03 mmol) in 20 mL of EtOH, add 65 mg of Pd / C, displace hydrogen 3 times, after reacting at 40 °C for 10 hours, filter the reaction solution through diatomaceous earth, and rotary evaporate the filtrate to obtain 0.45 g of a gray solid (crude yield: 50%). The crude product was directly used for the next reaction.

[0254] Step 7: Preparation of tert-butyl 4-((6-((3-(((5-bromopentyl)oxy)-4-morpholinophenyl)amino)-5-carbamoyl-3-ethylpyrazin-2-yl)amino)piperidine-1-carboxylate (Compound 8)

[0255]

[0256] Dissolve Compound 7 (0.45 g, 0.84 mmol) in 10 mL of DMF, add K2CO3 (0.28 g, 2.06 mmol) and 1,5-dibromopentane (0.28 g, 1.24 mmol), react at 70 °C for 4 hours, rotary evaporate the reaction solution, and obtain 0.2 g of a yellow solid by column chromatography (yield: 56%).

[0257] 11H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 7.54 (s, 1H), 7.27 - 7.24 (m, 2H), 7.05 (d, J = 2.3 Hz, 1H), 6.83 (d, J = 8.6 Hz, 1H), 6.74 (d, J = 7.4 Hz, 1H), 4.11 - 4.04 (m, 1H), 3.99 (t, J = 5.9 Hz, 4H), 3.72 (t, J = 4.5 Hz, 4H), 3.56 (t, J = 6.6 Hz, 2H), 2.93 (t, J = 4.6 Hz, 4H), 2.78 (s, 2H), 2.60 - 2.54 (m, 2H), 1.91 - 1.85 (m, 4H), 1.79 - 1.72 (m, 2H), 1.62 - 1.54 (m, 2H), 1.51 - 1.45 (m, 2H), 1.42 (s, 9H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 689.0 [M-H] - .

[0258] Step 8: 3 3 -Ethyl-5 4 -Morpholinyl-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -Carboxamide (Compound ZX27-8) Preparation

[0259]

[0260] Dissolve compound 8 (0.2 g, 0.29 mmol) in 10 mL of DCM, add 3 mL of TFA and react at room temperature for 2 hours. After extraction 3 times with DCM and saturated NaHCO3 solution, the organic layer is dried over anhydrous Na2SO4 and then evaporated to dryness to obtain a crude yellowish-brown solid. Directly dissolve this crude product in 60 mL of DMF, add potassium carbonate (60 mg, 0.44 mmol), react at 65 °C for 4 h, evaporate the reaction solution to dryness, and obtain 30 mg of yellow solid by column chromatography (total yield of two steps: 20%).

[0261] 11H NMR (400 MHz, DMSO-d6) δ 11.35 (s, 1H), 7.99 (s, 1H), 7.56 (s, 1H), 7.25 (s, 1H), 7.17 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 8.4 Hz, 1H), 6.58 (d, J = 7.9 Hz, 1H), 4.27 (s, 2H), 3.99 (s, 1H), 3.73 (s, 4H), 3.27 - 3.17 (m, 2H), 3.05 (s, 2H), 2.94 - 2.86 (m, 6H), 2.56 (q, J = 7.5 Hz, 2H), 2.02 - 1.87 (m, 4H), 1.70 (s, 6H), 1.18 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 508.1 [M-H] - .

[0262] Example 15: 3 3 -Ethyl-5 4 -Morpholinyl-6,9-dioxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -Carboxamide (Compound ZX31-9) Preparation

[0263]

[0264] The synthesis method is as in Example 14.

[0265] 1 1H NMR (400 MHz, DMSO-d6) δ 11.32 (s, 1H), 8.27 (d, J = 2.5 Hz, 1H), 7.58 (s, 1H), 7.30 - 7.25 (m, 2H), 6.94 (d, J = 8.6 Hz, 1H), 6.63 (dd, J = 8.6, 2.6 Hz, 1H), 4.34 (t, J = 3.4 Hz, 2H), 4.21 - 4.17 (m, 1H), 3.88 (d, J = 4.7 Hz, 2H), 3.79 (d, J = 3.4 Hz, 2H), 3.72 (t, J = 4.5 Hz, 4H), 3.61 (s, 2H), 3.26 (s, 4H), 2.96 - 2.93 (m, 4H), 2.61 - 2.56 (m, 2H), 2.06 - 1.99 (m, 2H), 1.84 - 1.81 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 510.3 [M-H] - .

[0266] Example 16: 3 3-Ethyl-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 Preparation of -carboxamide (Compound ZX34-8)

[0267]

[0268] The synthesis method is as in Example 14.

[0269] 1 H NMR (400 MHz, DMSO-d6) δ 11.31 (s, 1H), 8.07 (s, 1H), 7.60 (s, 1H), 7.30 (s, 1H), 7.24 (d, J = 7.2 Hz, 1H), 7.18 (t, J = 8.1 Hz, 1H), 6.55 (dd, J = 8.2, 2.4 Hz, 1H), 6.47 (dd, J = 8.0, 2.0 Hz, 1H), 4.00 (t, J = 4.9 Hz, 2H), 3.26 - 3.21 (m, 3H), 3.07 (s, 2H), 2.58 (q, J = 7.4 Hz, 2H), 2.51 (s, 2H), 2.05 - 2.01 (m, 2H), 1.86 - 1.75 (m, 8H), 1.19 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 423.1 [M-H] - .

[0270] Example 17: 3 3 -Isopropyl-5 4 -Morpholinyl-6,9-dioxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzonacycloundecane-3 5 Preparation of -carboxamide (Compound ZX42-9)

[0271]

[0272] The synthesis method is as in Example 14.

[0273] 11H NMR (400 MHz, DMSO-d6) δ 11.30 (s, 1H), 8.26 (d, J = 2.6 Hz, 1H), 7.55 (d, J = 3.3 Hz, 1H), 7.30 (dd, J = 9.3, 5.3 Hz, 2H), 6.93 (d, J = 8.6 Hz, 1H), 6.63 (dd, J = 8.6, 2.6 Hz, 1H), 4.34 (d, J = 3.7 Hz, 2H), 4.25 - 4.16 (m, 1H), 3.88 (d, J = 3.8 Hz, 2H), 3.79 (d, J = 4.0 Hz, 2H), 3.72 (t, J = 4.5 Hz, 4H), 3.62 (s, 2H), 3.27 (s, 4H), 3.18 - 3.11 (m, 1H), 2.94 (t, J = 4.4 Hz, 4H), 2.06 - 2.02 (m, 2H), 1.87 - 1.79 (m, 2H), 1.16 (d, J = 6.5 Hz, 6H). LC-MS (ESI) m / z 524.1 [M-H] - .

[0274] Example 18: (1 3 S)-3 3 -ethyl-5 4 -morpholino-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(3,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX32-9) Preparation

[0275]

[0276] The synthesis method was as in Example 14.

[0277] 1 1H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 7.89 (s, 1H), 7.52 (s, 1H), 7.20 (s, 1H), 6.79 (d, J = 8.4 Hz, 1H), 6.63 (d, J = 8.8 Hz, 1H), 6.42 (d, J = 6.5 Hz, 1H), 4.26 (s, 1H), 4.16 - 4.11 (m, 1H), 4.03 - 3.98 (m, 1H), 3.71 (t, J = 4.6 Hz, 4H), 3.06 - 3.03 (m, 1H), 2.99 - 2.83 (m, 5H), 2.59 - 2.53 (m, 2H), 2.41 - 2.25 (m, 2H), 2.03 - 1.84 (m, 2H), 1.70 - 1.67 (m, 4H), 1.54 - 1.43 (m, 6H), 1.18 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 508.3 [M-H]- .

[0278] Example 19: (1 3 (R)-3 3 -ethyl-5 4 -morpholino-6-oxa-2,4-diaza-3(2,6)-pyrazin-1(3,1)-piperidin-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX35-9) Preparation

[0279]

[0280] The synthesis method is as in Example 14.

[0281] 1 H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 7.89 (d, J = 2.4 Hz, 1H), 7.52 (d, J = 3.4 Hz, 1H), 7.20 (d, J = 3.3 Hz, 1H), 6.79 (d, J = 8.4 Hz, 1H), 6.62 (d, J = 8.8 Hz, 1H), 6.42 (dd, J = 8.4, 2.3 Hz, 1H), 4.27 - 4.24 (m, 1H), 4.16 - 4.11 (m, 1H), 4.03 - 3.97 (m, 1H), 3.71 (t, J = 4.6 Hz, 4H), 3.04 (dd, J = 10.3, 4.1 Hz, 1H), 2.98 - 2.83 (m, 5H), 2.59 - 2.53 (m, 2H), 2.40 - 2.36 (m, 1H), 2.28 - 2.24 (m, 1H), 1.93 - 1.86 (m, 1H), 1.81 - 1.67 (m, 6H), 1.55 - 1.40 (m, 5H), 1.17 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 508.3 [M-H] - .

[0282] Example 20: (1 3 (R)-3 3 -ethyl-5 4 -morpholino-6-oxa-2,4-diaza-3(2,6)-pyrazin-1(3,1)-pyrrolidin-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX37-9) Preparation

[0283]

[0284] The synthesis method is as in Example 14.

[0285] 11H NMR (400 MHz, DMSO-d6) δ 11.34 (s, 1H), 8.07 (d, J = 2.4 Hz, 1H), 7.56 (d, J = 3.3 Hz, 1H), 7.23 (d, J = 3.4 Hz, 1H), 6.91 (d, J = 7.2 Hz, 1H), 6.82 (d, J = 8.5 Hz, 1H), 6.48 (dd, J = 8.5, 2.4 Hz, 1H), 4.50 - 4.41 (m, 1H), 4.16 - 4.11 (m, 1H), 3.88 - 3.83 (m, 1H), 3.71 - 3.63 (m, 5H), 3.07 - 3.02 (m, 2H), 2.94 (d, J = 5.8 Hz, 1H), 2.84 - 2.79 (m, 2H), 2.61 - 2.56 (m, 2H), 2.51 (s, 1H), 2.34 - 2.25 (m, 2H), 2.18 - 1.99 (m, 2H), 1.91 - 1.81 (m, 2H), 1.68 - 1.59 (m, 2H), 1.48 - 1.39 (m, 3H), 1.18 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 494.1 [M-H]-.

[0286] Example 21: (1 3 (S)-3 3 -ethyl-5 4 -morpholinyl-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(3,1)-pyrrolidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX36-9) Preparation

[0287]

[0288] The synthesis method is as in Example 14.

[0289] 11H NMR (400 MHz, DMSO-d6) δ 11.34 (s, 1H), 8.07 (s, 1H), 7.56 (d, J = 3.4 Hz, 1H), 7.24 (d, J = 3.3 Hz, 1H), 6.92 (d, J = 7.2 Hz, 1H), 6.82 (d, J = 8.5 Hz, 1H), 6.48 (dd, J = 8.5, 2.4 Hz, 1H), 4.51 - 4.41 (m, 1H), 4.16 - 4.11 (m, 1H), 3.89 - 3.83 (m, 1H), 3.71 - 3.63 (m, 5H), 3.07 - 3.02 (m, 2H), 2.97 (d, J = 9.0 Hz, 1H), 2.83 - 2.79 (m, 2H), 2.61 - 2.56 (m, 2H), 2.53 (s, 1H), 2.36 - 2.29 (m, 2H), 2.15 - 2.00 (m, 2H), 1.94 - 1.81 (m, 2H), 1.70 - 1.60 (m, 2H), 1.48 - 1.44 (m, 3H), 1.18 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 494.1 [M-H] - .

[0290] Example 22: 3 5 -Ethyl-1 4 -Morpholinyl-12-oxa-2,4-diaza-3(2,6)-pyrazine-6(4,1)-piperidine-1(1,3)-benzocyclododecanone 3 3 -Formamide (Compound ZX33-9) Preparation

[0291]

[0292] The synthesis method is as in Example 14.

[0293] 11H NMR (400 MHz, Chloroform-d) δ 10.13 (s, 1H), 7.46 (s, 1H), 6.99 - 6.96 (m, 2H), 6.88 (d, J = 8.5 Hz, 1H), 5.21 (s, 1H), 4.88 (d, J = 6.6 Hz, 1H), 4.19 - 4.16 (m, 2H), 3.87 (t, J = 4.6 Hz, 4H), 3.46 (s, 2H), 3.10 (t, J = 4.6 Hz, 4H), 2.69 (s, 4H), 2.55 - 2.49 (m, 2H), 2.45 (s, 1H), 1.83 - 1.80 (m, 4H), 1.67 - 1.64 (m, 2H), 1.57 - 1.52 (m, 2H), 1.45 - 1.39 (m, 2H), 1.32 - 1.29 (m, 3H), 0.91 - 0.85 (m, 2H). LC-MS (ESI) m / z 522.3 [M-H]-.

[0294] Example 23: 3 3 -Ethyl-5 4 -Morpholinyl-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncyclododecane-3 5 -Formamide (Compound ZX45-9) Preparation

[0295]

[0296] The synthesis method is as in Example 14.

[0297] 1 1H NMR (400 MHz, DMSO-d6) δ 11.28 (s, 1H), 7.83 (s, 1H), 7.58 (s, 1H), 7.28 (s, 1H), 7.06 (d, J = 4.8 Hz, 1H), 6.91 (d, J = 8.4 Hz, 1H), 6.59 (d, J = 7.7 Hz, 1H), 4.16 - 4.13 (m, 2H), 4.01 (s, 1H), 3.75 (s, 4H), 3.42 (s, 2H), 3.14 (s, 2H), 2.94 (s, 4H), 2.59 (q, J = 7.4, 6.9 Hz, 2H), 2.14 (s, 2H), 1.94 - 1.86 (m, 5H), 1.76 (s, 2H), 1.67 - 1.62 (m, 5H), 1.19 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z522.3 [M-H] - .

[0298] Example 24: 3 3 -Ethyl-5 4-Morpholino-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzocyclotridecane-3 5 Preparation of -formamide (Compound ZX46-9)

[0299]

[0300] The synthesis method is as in Example 14.

[0301] 1 H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.74 (d, J = 2.5 Hz, 1H), 7.57 (d, J = 3.3 Hz, 1H), 7.27 (d, J = 3.3 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 6.59 (dd, J = 8.5, 2.5 Hz, 1H), 4.10 (d, J = 5.3 Hz, 2H), 4.01 (s, 1H), 3.75 (t, J = 4.4 Hz, 4H), 3.40 (s, 2H), 3.10 (s, 4H), 2.96 (t, J = 4.3 Hz, 4H), 2.60 (q, J = 7.4 Hz, 2H), 2.12 (d, J = 9.6 Hz, 2H), 1.98 - 1.89 (m, 2H), 1.77 - 1.72 (m, 2H), 1.68 - 1.63 (m, 4H), 1.49 (s, 4H), 1.19 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 536.1 [M-H] - .

[0302] Example 25: 3 3 -Ethyl-9-methyl-5 4 -Morpholino-6-oxa-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzocycloundecane-3 5 -Carboxamide (Compound ZX44-9) Preparation

[0303]

[0304] The synthesis method is as in Example 14.

[0305] 11H NMR (400 MHz, DMSO-d6) δ 11.39 (s, 1H), 8.08 (s, 1H), 7.59 (s, 1H), 7.30 (s, 1H), 7.25 (d, J = 6.9 Hz, 1H), 6.95 (d, J = 8.4 Hz, 1H), 6.62 (d, J = 8.3 Hz, 1H), 4.34 - 4.28 (m, 2H), 4.16 (s, 1H), 3.79 - 3.71 (m, 4H), 3.44 (s, 2H), 3.25 - 3.10 (m, 6H), 2.75 (d, J = 8.6 Hz, 2H), 2.58 (q, J = 7.7 Hz, 2H), 2.21 - 1.98 (m, 6H), 1.85 - 1.69 (m, 2H), 1.43 (t, J = 11.9 Hz, 1H), 1.19 (t, J = 7.1 Hz, 3H), 0.97 (d, J = 6.2 Hz, 3H). LC-MS (ESI) m / z 522.1 [M-H] - .

[0306] Example 26: 3 3 -Ethyl-5 4 -(4-Methylpiperazin-1-yl)-6,9-dioxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX49-9) Preparation

[0307]

[0308] The synthesis method was as in Example 14.

[0309] 11H NMR (400 MHz, DMSO-d6) δ 11.36 (s, 1H), 8.28 (d, J = 2.5 Hz, 1H), 7.60 (d, J = 3.3 Hz, 1H), 7.32 (d, J = 4.6 Hz, 2H), 6.97 (d, J = 8.5 Hz, 1H), 6.64 (dd, J = 8.6, 2.5 Hz, 1H), 4.32 (s, 2H), 4.25 - 4.22 (m, 1H), 3.93 (t, J = 4.8 Hz, 2H), 3.82 (t, J = 4.5 Hz, 2H), 3.76 - 3.69 (m, 2H), 3.48 (d, J = 11.9 Hz, 2H), 3.32 - 3.27 (m, 4H), 3.20 - 3.09 (m, 6H), 2.72 (s, 3H), 2.59 (q, J = 7.4 Hz, 2H), 2.12 - 2.09 (m, 2H), 1.89 (q, J = 13.4 Hz, 2H), 1.19 (t, J = 7.3 Hz, 3H). LC-MS (ESI) m / z 523.1 [M-H] - .

[0310] Example 27: 3 3 -Ethyl-5 4 -(4-Morpholinopiperidin-1-yl)-6,9-dioxo-2,4-diaza-3(2,6)-pyrazine-1(4,1)-piperidine-5(1,3)-benzoncycloundecane-3 5 -carboxamide (Compound ZX50-9) Preparation

[0311]

[0312] The synthesis method was as in Example 14.

[0313] 1 1H NMR (400 MHz, DMSO-d6) δ 11.25 (s, 1H), 8.27 (s, 1H), 7.53 (s, 1H), 7.22 (s, 1H), 7.14 (d, J = 8.3 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 6.57 (d, J = 8.2 Hz, 1H), 4.27 (s, 2H), 4.12 (s, 1H), 3.74 (s, 4H), 3.58 (s, 4H), 3.43 (s, 4H), 3.27 (s, 4H), 2.94 (s, 4H), 2.64 - 2.54 (m, 2H), 2.18 (s, 1H), 1.87 (d, J = 10.8 Hz, 4H), 1.65 (s, 2H), 1.49 - 1.33 (m, 4H), 1.17 (d, J = 7.5 Hz, 3H). LC-MS (ESI) m / z 593.0 [M-H] - .

[0314] Example 28: 3 3 -Ethyl-5 4 -(4-(4-Methylpiperazin-1-yl)piperidin-1-yl)-6,9-dioxo-2,4-diazolo-3(2,6)-pyrazin-1(4,1)-piperidin-3 5 Preparation of (1,3)-benzo

[11] annulene-35-carboxamide (Compound ZX51-9)

[0315]

[0316] The synthesis method is as in Example 14.

[0317] 1 H NMR (400 MHz, DMSO-d6) δ 11.30 (s, 1H), 8.26 (d, J = 2.5 Hz, 1H), 7.58 (d, J = 3.3 Hz, 1H), 7.28 (d, J = 7.6 Hz, 2H), 6.93 (d, J = 8.6 Hz, 1H), 6.61 (dd, J = 8.6, 2.6 Hz, 1H), 4.32 (d, J = 3.0 Hz, 2H), 4.26 - 4.17 (m, 1H), 3.90 (t, J = 4.7 Hz, 2H), 3.80 (t, J = 3.9 Hz, 2H), 3.66 (t, J = 9.9 Hz, 2H), 3.41 - 3.38 (m, 8H), 2.89 (s, 6H), 2.58 (q, J = 7.2 Hz, 5H), 2.50 (s, 3H), 2.08 (d, J = 12.8 Hz, 2H), 1.90 - 1.83 (m, 4H), 1.53 (s, 2H), 1.19 (t, J = 7.4 Hz, 3H). LC-MS (ESI) m / z 606.1 [M-H] - .

[0318] Example 29: 5 4 -(4-(Dimethylamino)piperidin-1-yl)-3 3 -Ethyl-6,9-dioxo-2,4-diaza-3(2,6)-pyrazin-1(4,1)-piperidin-5(1,3)-benzo

[11] annulene-3 5 -Carboxamide (Compound ZX53-9) Preparation

[0319]

[0320] The synthesis method is as in Example 14.

[0321] 11H NMR (400 MHz, Methanol-d4) δ 8.38 (s, 1H), 6.99 (d, J = 8.8 Hz, 1H), 6.68 (d, J = 8.6 Hz, 1H), 4.46 - 4.42 (m, 1H), 4.39 (s, 2H), 4.09 - 4.00 (m, 4H), 3.93 (s, 2H), 3.58 (d, J = 11.5 Hz, 4H), 3.48 (d, J = 4.9 Hz, 2H), 3.30 - 3.27 (m, 1H), 2.94 (s, 6H), 2.71 (t, J = 11.8 Hz, 2H), 2.62 (q, J = 7.3 Hz, 2H), 2.35 (d, J = 13.5 Hz, 2H), 2.21 (d, J = 11.6 Hz, 2H), 2.06 - 1.83 (m, 4H), 1.27 (d, J = 7.0 Hz, 3H). LC-MS (ESI) m / z 551.0 [M-H] - .

[0322] Example 30: Compound against FLT3 D835Y kinase IC 50 test

[0323] Kinase activity detection: The Eu kinase binding assay is based on the binding and displacement of a proprietary Alexa 647-labeled ATP-competitive kinase inhibitor scaffold (kinase tracer) to the kinase. The binding of the tracer to the kinase is detected by an Eu (europium)-labeled anti-tag antibody that also binds to the kinase. The binding of the tracer and the antibody to the kinase results in a high degree of fluorescence resonance energy transfer (FRET), converting the Eu donor fluorophore to an Alexa 647 acceptor fluorophore. Conversely, the binding of the inhibitor to the kinase competes with the binding of the tracer, resulting in the loss of FRET.

[0324] Enzymatic reaction: In a 384-well plate, add 5 μL of the enzyme-antibody mixture. Use an Echo550 ultra-micro liquid pipetting system to transfer 5 nL of the test compound (concentration gradient). After shaking, mixing, and centrifuging, add 5 μL of the tracer. After shaking, mixing, and centrifuging again, react at room temperature for 1 h.

[0325] Detection reaction: Add 2.5 μL of the reaction solution (Development Solution) (diluted 1:128) to each well and incubate at 37 °C in the dark for 1 h, then add 5 μL of the stop solution (Stop Reagent).

[0326] Plate reading: A multi-label microplate reader (Perkin Elmer EnVision Multimode Plate Reader) was used to detect the fluorescence signals (excitation wavelength: 340 nm, emission wavelengths: 665 nm, 615 nm).

[0327] Calculation: The inhibition rate of each well was calculated based on the fully active wells and the control signal wells. The data analysis method was as follows:

[0328] Phosphorylation ratio = 1 - {[(Emission ratio × F100% - C100%)] / [C0% - C100% + Emission ratio × (F100% - F0%)]} × 100;

[0329] Inhibition rate = 100 × (1 - Compound phosphorylation ratio / Negative control phosphorylation ratio).

[0330] IC 50 values were calculated using medical drawing software (GraphPad Prism 5.0).

[0331] The results of the kinase activity test are shown in Table 1.

[0332] Example 31: The effect of the compound on FLT3 D835Y / F691L kinase IC 50 testing

[0333] Kinase activity detection: Using the Z′-LYTE TM technology (fluorescence detection, enzyme-coupled form, based on the difference in sensitivity of phosphorylated and non-phosphorylated polypeptides to proteolytic cleavage), the principle of fluorescence resonance energy transfer (FRET) was adopted, and Z′-LYTE TM FRET peptide substrates were used to detect the inhibitory activity of the compound against FLT3 D835Y / E69L kinase in a secondary reaction.

[0334] Enzymatic reaction: In a 384-well plate, add 5 μL of the enzyme-non-phosphorylated substrate system [50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.5, 0.01% BRIJ-35, 10 mM magnesium chloride (MgCl2), 1 mM ethylene glycol bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 2 μM Tyr 02 non-phosphorylated substrate]. Use the Echo550 ultra-micro liquid transfer system to transfer 5 nL of the test compound (concentration gradient). After shaking at room temperature for 15 min, use the Echo550 ultra-micro liquid transfer system to transfer 250 nL of ATP (final concentration of 500 μM) respectively. After shaking and mixing, centrifuge and react at 30 °C in the dark for 1.5 h. At the same time, prepare 5 μL of the phosphorylated substrate system wells [50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.5, 0.01% BRIJ-35, 10 mM magnesium chloride (MgCl2), 1 mM ethylene glycol bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 2 μM Tyr 02 phosphorylated peptide substrate] as a control.

[0335] Detection reaction: Add 2.5 μL of the reaction solution (Development Solution) (diluted 1:128) to each well and incubate at 37 °C in the dark for 1 h, then add 5 μL of the stop solution (Stop Reagent).

[0336] Plate reading: Use a multi-label microplate reader (Perkin Elmer EnWision Multimode Plate Reader) to detect the fluorescence signal (excitation wavelength is 400 nm, emission wavelengths are 460 nm and 535 nm).

[0337] Calculation: Calculate the inhibition rate of each well through the fully active wells and the control signal wells. The data analysis method is as follows:

[0338] Phosphorylation ratio = 1 - {[(emission ratio × F100% - C100%) / [C0% - C100% + emission ratio × (F100% - F0%)]]} × 100;

[0339] Inhibition rate = 100 × (1 - compound phosphorylation ratio / negative control phosphorylation ratio).

[0340] IC 50 values are calculated using medical drawing software (GraphPad Prism5.0).

[0341] The results of the kinase activity test are shown in Table 1.

[0342] Example 32: Study on the cell proliferation inhibitory activity based on Ba / F3-FLT3 stable cell line

[0343] The BaF3 cells (mouse pre-B cells) used in this experiment were purchased from the Japanese Cell Bank. The monoclonal stable strains of BaF3-ITD, BaF3-ITD-D835Y, and BaF3-ITD-F691L were all constructed by our laboratory and were fully verified by experiments such as positive drug activity, protein expression, and gene sequencing.

[0344] The brief steps for constructing the stable strains are as follows: Construct pCDNA3.1(+) plasmid vectors carrying genes such as ITD, ITD-D835Y, and ITD-F691L; use Cell Line Kit V to electrotransfer the plasmids into Ba / F3 cells; 48 hours after electrotransfer, add geneticin (G418) at a final concentration of 1000 μg / ml and screen for two weeks, then remove interleukin 3 (IL3) and continue screening to obtain polyclonal stable strains; then select monoclonal strains by the limiting dilution method; and further identify the stable strains using positive drugs, Western Blot (WB), and gene sequencing; the monoclonal strains that are fully verified can be used for the study of the inhibitory activity of inhibitors on cell proliferation.

[0345] Study on the inhibitory activity of cell proliferation: Inoculate cells in the logarithmic growth phase at 8000 - 12000 cells / well into a 96-well plate. The next day, add inhibitors at different concentrations (0 - 10 μM) and continue culturing for 72 hours; then add 10 μL of Cell Counting Kit-8 cell counting reagent (CCK-8 reagent) to each well and continue incubating for 1 - 3 hours; then measure the absorbance values at 450 nm and 650 nm using a microplate reader. Use medical drawing software (GraphPad Prism 8.0.0) to calculate the half-maximal inhibitory concentration (IC 50 50

[0346] The test results are shown in Table 1.

[0347] Table 1 Test results of the kinase activity and cell proliferation inhibitory activity of the compounds (IC 50 50

[0348]

[0349]

[0350] IC 50 50

[0351] As can be seen from the data in Table 1, the novel cyclic compound of the present invention has strong inhibitory activity against the cell proliferation of FLT3 mutant kinase and Ba / F3-FLT3 stable strain.

[0352] Example 33: Study on the inhibitory activity of cell proliferation based on human tumor cell lines

[0353] The human acute monocytic leukemia cell line MV4-11 (containing FLT3-ITD insertion mutation) was purchased from the American Type Culture Collection (ATCC). The cells were seeded in 96-well plates at 18,000 cells per well using complete medium (IMDM + 10% FBS + 1% Penicillin-streptomycin). The next day, different concentrations of the inhibitor (0 - 10 μM) were added and the cells were cultured for another 72 hours; then 10 μL of Cell Counting Kit-8 cell counting reagent (CCK-8 reagent) was added to each well and incubated for 1 - 3 hours; then the absorbance values at 450 nm and 650 nm were measured using a BioTeK Synergy H1 Hybrid Reader microplate reader. The IC 50 value was calculated by non-linear fitting of the curve using GraphPad Prism 8.0.1 software. Each measurement was repeated at least three times.

[0354] The test results are shown in Table 2.

[0355] Table 2 Test results of the inhibitory activity of the compound against the cell proliferation of MV4-11 cells (IC 50 : nM)

[0356] Compound Number ZX18-8 ZX27-8 ZX31-9 MV4-11 *** * *

[0357] As can be seen from the data in Table 2, the compound of the present invention has strong inhibitory activity against the cell proliferation of MV4-11 cells.

[0358] Example 34: Pharmacokinetic evaluation

[0359] Pharmacokinetic and oral bioavailability tests were performed on Sprague-Dawley rats. According to the drug solubility, the drug was administered orally and intravenously as a single dose. After collecting animal blood samples at different time points (oral: 0.25, 0.5, 1, 2, 4, 6, 8, 24 h; intravenous: 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 h), the blood samples were analyzed using HPLC-MS method, and the pharmacokinetic parameters were calculated using WinNonlin7.0 software, including the half-life (T1 / 2), the maximum plasma concentration (Cmax), the time to reach the maximum concentration (Tmax), the area under the plasma concentration-time curve (AUC), the bioavailability (BA), and other pharmacokinetic data.

[0360] The pharmacokinetic data results of compound ZX31-9 are shown in Table 3 as follows:

[0361]

[0362]

[0363] As can be seen from the data in Table 3, compound ZX31-9 has good oral absorption properties.

[0364] Example 35: In vivo xenograft model study

[0365] The in vivo tumor growth inhibitory activity of the present invention was studied using the Ba / F3-FLT3-ITD model cell line containing the D835Y point mutation in a xenograft mouse model. All male Balb / C nude mice at 4-5 weeks of age were housed in plastic cages with corncobs (3-4 mice / cage), maintained in a pathogen-free facility with a 12-hour light / dark cycle (20-25 °C, 30-70% humidity). A tumor model was established by subcutaneous injection of the Ba / F3-FLT3-ITD-D835Y model cell suspension. When the average tumor volume reached approximately 100 mm 3 (about 5 days after tumor implantation), the tumor-bearing mice were randomly divided into 3 groups (control group and dosing group, 7-8 mice in each group). Subsequently, the dosing group was treated with 10 mg / kg or 30 mg / kg of compound ZX31-9 daily (administered by gavage after dissolving and suspending with 0.5% methylcellulose as a vehicle), and the control group mice were gavaged with 0.5% methylcellulose as a vehicle for 12 days; the body weight and tumor volume of the mice were measured every two days. When appropriate, the tumor growth inhibition rate (TGI) of each group was calculated using the following formula: TGI = (1 - RTV 治疗组 / RTV 对照组 ) * 100%, RTV 相对肿瘤体积 = Vt 肿瘤体积 / V0 开始肿瘤体积 .

[0366] The test results are as Figure 1 shown. As can be seen from Figure 1 the data, at the doses of 10 mg / kg and 30 mg / kg, compound ZX31-9 dose-dependently inhibited tumor growth with TGI (tumor growth inhibition rate) of 88.2% and 51.6% respectively on the sixth day, and no obvious weight loss was observed throughout the process. These results indicate that compound ZX31-9 has significant anti-cancer efficacy and no potential toxicity in the Ba / F3-FLT3-ITD-D835Y xenograft tumor model.

[0367] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the following embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope recorded in this specification.

[0368] The above-described embodiments merely represent several implementation manners of the present invention. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent for the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention shall be subject to the appended claims.

Claims

1. A cyclic compound having the structure shown in formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof: Among them, E, Z, and Q are each independently selected from: CR 1 or N; Each R 1 is independently selected from: H, halogen, R 2 substituted or unsubstituted C1-C6 alkyl, R 2 substituted or unsubstituted C1-C6 alkoxy, R 2 substituted or unsubstituted C3-C8 cycloalkyl; Each R 2 is independently selected from: H, halogen, C1-C6 alkyl, C1-C6 alkoxy; X, Y, K, D are each independently selected from: -O-, -C(R 3 R 3 )-, -N(R 3 )C(R 3 R 3 )-, -N(R 3 )-, -S-, -S(=O)-, -S(O)2-, -C(=O)-, -(C=O)N(R 3 )-; Each R 3 is independently selected from: H, C1-C6 alkyl; L is selected from: R 4 substituted or unsubstituted C2-C 12 alkylene, R 4 substituted or unsubstituted C2-C 12 oxaalkylene, R 4 substituted or unsubstituted C2-C 12 thiaalkylene, R 4 substituted or unsubstituted C2-C 12 unsaturated hydrocarbon radical; Each R 4 is independently selected from: H, halogen, C1-C6 alkyl; A is selected from: R 5 substituted or unsubstituted C3-C6 cycloalkyl, R 5 substituted or unsubstituted 4-7 membered saturated nitrogen-containing heterocyclic group; Each R 5 is independently selected from: H, halogen, nitro, hydroxy, cyano, C1-C6 alkyl, C1-C6 alkoxy; B is selected from: H, halogen, nitro, hydroxy, cyano, R 6 substituted or unsubstituted C1-C 20 alkyl, R 6 substituted or unsubstituted C1-C 20 alkoxy, R 6 substituted or unsubstituted C3-C 12 cycloalkyl, R 6 substituted or unsubstituted 3- to 12-membered heterocyclic group, R 6 substituted or unsubstituted 5- to 10-membered heteroaryl, -NR 7 R 8 ; Each R 6 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy, R 9 substituted or unsubstituted 3- to 8-membered heterocyclic group, -NR 3 R 3 ; R 7 and R 8 are each independently selected from: H, R 6 substituted or unsubstituted C1-C6 alkyl, R 6 substituted or unsubstituted 3- to 12-membered heterocyclic group, or R 7 and R 8 together with the nitrogen atom to which it is attached form R 6 substituted or unsubstituted 3- to 12-membered heterocyclic group; Each R 9 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C6 alkyl, C1-C6 alkoxy.

2. The cyclic compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, Having the structure shown in formula (II):

3. The cyclic compound according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein, R 1 Selected from: H, C1-C6 alkyl; preferably methyl, ethyl, n-propyl, isopropyl.

4. The cyclic compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, X, Y, K, and D are each independently selected from: -O-, -N(R 3 )-, -N(R 3 )C(R 3 R 3 )-, -(C=O)N(R 3 )-, -C(=O)-, -C(R 3 R 3 )-; R 3 is selected from: H, C1-C3 alkyl.

5. The cyclic compound according to claim 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, Both X and Y are -N(H)-; and / or, K is selected from: -O-, -(C=O)N(H)-; and / or, D is selected from: -C(=O)-, -CH2-, -N(H)-.

6. The cyclic compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, L is selected from: R 4 Substituted or unsubstituted C3-C8 alkylene, -(CR 2 R 2 ) n -O-(CR 2 R 2 ) m -, -(CR 2 R 2 ) n -S-(CR 2 R 2 ) m -, R 4 Substituted or unsubstituted C3-C8 unsaturated hydrocarbon group; Wherein, n and m are each independently selected from positive integers between 1 and 7, and n + m is not greater than 8; Each R 2 is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl; Each R 4 is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl.

7. The cyclic compound according to claim 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, L is selected from: -(CH2) a -, -(CH2) n -O-(CH2) m -, -(CH2) b -CH(R 4 )-(CH2) c -; Wherein, a is selected from: 3, 4, 5, 6, 7, 8; n and m are each independently selected from: 1, 2, 3; b and c are each independently selected from: 1, 2, 3; R 4 Selected from: methyl, ethyl.

8. The cyclic compound according to claim 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein, L is selected from: -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)2-O-(CH2)2-, -(CH2)2-CH(CH3)-(CH2)2-.

9. The cyclic compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that A is selected from: a 5- to 6-membered saturated nitrogen-containing heterocyclic group, and the nitrogen atom is connected to D.

10. The cyclic compound according to claim 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein, A is selected from:

11. The cyclic compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, B is selected from: H, R 6 substituted or unsubstituted C1-C6 alkyl, R 6 substituted or unsubstituted C1-C6 alkoxy, R 6 substituted or unsubstituted C3-C8 cycloalkyl, R 6 substituted or unsubstituted 3-8 membered heterocyclic group, R 6 substituted or unsubstituted 5-10 membered heteroaryl, -NR 7 R 8 ; Each R 6 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C3 alkyl, C1-C3 alkoxy, R 9 substituted or unsubstituted 4-6 membered heterocyclic group, -NR 3 R 3 ; R 7 and R 8 are each independently selected from: H, R 6 substituted or unsubstituted C1-C3 alkyl, R 6 substituted or unsubstituted 3- to 8-membered heterocyclic group, or R 7 and R 8 together with the nitrogen atom to which it is attached form a R 6 substituted or unsubstituted 3- to 8-membered heterocyclic group; Each R 9 is independently selected from: H, halogen, nitro, hydroxy, cyano, mesyl, C1-C3 alkyl, C1-C3 alkoxy; Each R 3 is independently selected from: H, C1-C3 alkyl.

12. The cyclic compound according to claim 11, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, B is selected from: H, -NR 7 R 8 ; R 7 and R 8 are each independently selected from: H, R 6 substituted or unsubstituted C1-C3 alkyl, R 6 substituted or unsubstituted 5- to 6-membered heterocyclic group, or R 7 and R 8 together with the nitrogen atom to which it is attached form R 6 substituted or unsubstituted 5- to 6-membered heterocyclic group; Each R 6 is independently selected from: H, hydroxyl, methyl, ethyl, propyl, methoxy, ethoxy, R 9 a substituted or unsubstituted 4- to 6-membered heterocyclic group, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, dipropylamino; Each R 9 is independently selected from: H, fluorine, chlorine, bromine, methyl, ethyl, propyl, methoxy, ethoxy.

13. The cyclic compound according to claim 11, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, B is selected from: H, methyl, ethyl, methoxy, ethoxy, cyclopentyl, cyclohexyl, Each R 6 is independently selected from: H, methyl, ethyl, propyl, dimethylamino, and methanesulfonyl; R 9 Selected from: H, methyl, ethyl, propyl, dimethylamino, methanesulfonyl.

14. The cyclic compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that Both X and Y are -N(H)-, K is -O-, and D is -CH2-; L is selected from: -(CH2)4-, -(CH2)2-O-(CH2)2-, -(CH2)2-CH(CH3)-(CH2)2-; A is B is selected from: -NR 7 R 8 ; R 7 and R 8 are each independently selected from: methyl, ethyl, propyl, methyl substituted with dimethylamino, ethyl substituted with dimethylamino, propyl substituted with dimethylamino, or R 7 and R 8 together with the nitrogen atom to which it is attached form a R 6 substituted or unsubstituted 5- or 6-membered heterocyclic group; Each R 6 is independently selected from: H, methyl, ethyl, propyl, R 9 a substituted or unsubstituted 4- to 6-membered heterocyclic group, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, dipropylamino; Each R 9 is independently selected from: H, methyl, ethyl, propyl.

15. The cyclic compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that, The cyclic compound is selected from the following compounds:

16. Use of the cyclic compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof in the preparation of an FLT3 inhibitor; preferably, the FLT3 is a mutated FLT3, preferably an FLT3-ITD mutation, an FLT3-TKD mutation; preferably FLT3 ITD mutation, FLT3 D835Y mutation, FLT3 D835Y / F691L mutation, FLT3 ITD / D835Y mutation, FLT3 ITD / F691L mutation.

17. Use of the cyclic compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, in the preparation of a drug for preventing and / or treating a disease mediated by FLT3; the disease is preferably a malignant blood disease; the malignant blood disease is preferably acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome.

18. A pharmaceutical composition for preventing and / or treating malignant hematological diseases, characterized in that, Prepared from an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient comprises the cyclic compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof.

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