Benzimidazole compound as well as preparation method and application thereof

Abstract

The invention discloses a benzimidazole compound as shown in a formula (I), and a preparation method and application thereof. In particular, the aforementioned compounds can be used to treat diseases associated with the biological activity of MNK1 and / or MNK2.

Description

A benzimidazole compound, its preparation method and application Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a benzimidazole compound, its preparation method, and its application. Background Technology

[0002] Mitogen-activated protein kinase-interacting kinases (MNKs) are a class of protein serine / threonine kinases comprising two subtypes, MNK1 and MNK2, encoded by the MKNK1 and MKNK2 genes, respectively. MNK1 and MNK2 are widely distributed and universally expressed in various tissues. Their activity can be regulated by extracellular signal-regulated kinases (ERKs) or p38 mitogen-activated protein kinase (MAPK), and can also be regulated by p21-activated protein kinase 2 (PAK2) and the mammalian target of rapamycin (mTOR).

[0003] Substrates for MNK phosphorylation modification include eukaryotic initiation factor 4E (eIF4E), heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), cytoplasmic phospholipase A2 (cPLA2), Sprouty 2 (Spry 2), and polypyrimidine tract binding protein-associated splicing factor (PSF), among which hnRNPA1 and eIF4E are the most typical.

[0004] hnRNPA1 is a nuclear protein that travels between the nucleus and cytoplasm, is involved in mRNA translation and turnover, and links MNKs to the mediation and regulation of inflammatory responses.

[0005] eIF4E is an mRNA cap-binding protein. Phosphorylation of eIF4E affects mRNA translation and its export from the nucleus. mRNA translation is crucial for general life activities, proliferation, and neoplastic diseases. MNK1 / 2 and its substrate eIF4E, often referred to as the MNK1 / 2-eIF4E axis, play a key role in the regulation of protein synthesis. Dysregulation of this pathway is associated with metabolic disorders and inflammation, autism spectrum disorder (ASD), infectious diseases, and malignant tumors. MNK phosphorylation of eIF4E affects the translation of an oncogenic subset of mRNAs. In tumor cells or tissues, high expression or overactivation of eIF4E leads to upregulation of the transcriptional levels of these mRNAs, playing a vital role in tumor cell survival and metastasis. Therefore, upregulation or overactivation of MNK kinases and eIF4E expression is often accompanied by poor prognosis. In the tumor immune microenvironment, the MNK1 / 2-eIF4E axis can influence the synthesis of various chemokines, cytokines, and immune checkpoint proteins, thereby regulating the immune response. Reports indicate that MNK inhibitors have no effect on PDL1 mRNA levels in tumor cells but significantly reduce the content of PDL1 on the surface of tumor cells, and can also reduce the expression of immune checkpoint proteins such as PL1, TIM3, and LAG3 on the surface of activated T cells. Furthermore, studies have found that MNK kinases play a role in compensatory pathways induced by various drugs, ultimately leading to drug resistance. Combining MNK inhibitors with these drugs can overcome this resistance pathway, producing a synergistic anti-tumor effect and effectively reversing the corresponding drug resistance. Of particular note is that knocking out or inhibiting MNK activity does not affect the growth of normal cells or the survival of animals; therefore, MNK inhibitors have significant application prospects and development value in tumor treatment. Summary of the Invention

[0006] The technical problem that the invention aims to solve:

[0007] Given the high application prospects and development value of MNK inhibitors in tumor treatment, this application provides a new compound that can serve as an MNK inhibitor.

[0008] This application also provides, in another respect, a pharmaceutical composition comprising this MNK inhibitor.

[0009] Solution for solving the problem:

[0010] In order to solve the above problems, the inventors of this application conducted in-depth research and found that benzimidazole compounds with specific structures can achieve the desired purpose, thus completing this invention.

[0011] This invention protects the following specific embodiments:

[0012] This invention provides a compound of formula (I), or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative thereof:

[0013] in,

[0014] R1 is selected from a 5-membered heteroaryl group; the heteroaryl group has 1-3 heteroatoms selected from N, O, and S; the heteroaryl group may optionally be surrounded by one or more R1 atoms. 1A Replace; R 1A Selected from C 1-6 Alkyl, -C(O)NR 1B R 1C 4-8 membered heterocyclic alkyl groups; the heterocyclic alkyl group contains one heteroatom O; R 1B and R 1C Each is independently selected from hydrogen and C. 1-6 Alkyl, halogen, cyano, amino, carboxyl, hydroxyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups;

[0015] R3 is independently selected from hydrogen, 4-12 membered heterocyclic alkyl groups, and 4-12 membered heterocyclic groups; the heterocyclic alkyl group or heterocyclic group contains 1-4 heteroatoms selected from N, O, and S; any carbon atom on the heterocyclic alkyl group or heterocyclic group can be oxidized; the heterocyclic alkyl group or heterocyclic group is unsubstituted or oxidized by one or more R3 groups. 3A Replace; each R 3A Each is independently selected from hydrogen and C. 1-6 Alkyl, halogen, cyano, amino, carboxyl, hydroxyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups;

[0016] R5 is independently selected from hydrogen, -C(O)NR 5B R 5C ;R 5B and R 5C Each is independently selected from hydrogen and C. 3-10 Cycloalkyl.

[0017] In one implementation, R3 and R5 are not both hydrogen.

[0018] In one embodiment, in the above-mentioned compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, R1 is selected from a 5-membered heteroaryl group; the heteroaryl group has two heteroatoms selected from N, O, and S; the heteroaryl group may optionally be surrounded by one or more R1 atoms. 1A replace.

[0019] Alternatively, in one embodiment, R1 is selected from a 5-membered heteroaryl group; the heteroaryl group has one or three heteroatoms selected from N, O, and S; the heteroaryl group may optionally be surrounded by one or more R1 atoms. 1A replace.

[0020] In a preferred embodiment, R1 is selected from one or more R1s. 1A Substituted pyrrole rings, pyrazole rings, imidazole rings, furan rings, thiophene rings, isoxazole rings, triazole rings, isothiazole rings, thiazole rings, and oxazole rings.

[0021] In a more preferred embodiment, R1 is selected from one or more R... 1A Substituted pyrazole ring, imidazole ring, thiazole ring.

[0022] Alternatively, in a more preferred embodiment, R1 is selected from one or more R... 1A Substituted pyrrole rings, furan rings, thiophene rings, isoxazole rings, triazole rings, isothiazole rings, and oxazole rings.

[0023] In a further preferred embodiment, R1 is selected from one or more R1s. 1A Replacement

[0024] In a further preferred embodiment, R1 is selected from one or more R1s. 1A Replacement

[0025] In one implementation scheme, each R 1A Each is independently selected from C 1-3 Alkyl, C 4-6 Alkyl, -C(O)NR 1B R 1C 4-6 membered heterocyclic alkyl groups, 7-8 membered heterocyclic alkyl groups; wherein the heterocyclic alkyl group contains one heteroatom O.

[0026] In a preferred embodiment, each R 1A Each is independently selected from methyl, ethyl, propyl, isopropyl, -C(O)NR 1B R 1C 4-5 membered heterocyclic alkyl, 6-7 membered heterocyclic alkyl; wherein the heterocyclic alkyl contains one heteroatom O.

[0027] Or in a preferred embodiment, each R 1A Each is independently selected from C 4-5 Alkyl, C6 alkyl, -C(O)NR 1B R 1C8-membered heterocyclic alkyl group; the heterocyclic alkyl group contains one heteroatom O.

[0028] In a further preferred embodiment, each R 1A Each is independently selected from methyl, -C(O)NR 1B R 1C A 6-membered heterocyclic alkyl group; wherein the heterocyclic alkyl group contains one heteroatom O.

[0029] In one implementation, R 1B R 1C Each is independently selected from hydrogen and C. 1-3 Alkyl, C 4-6 Alkyl, fluorine, chlorine, bromine, iodine, cyano, amino, carboxyl, hydroxyl, nitro, C 1-3 Alkoxy, C 4-6 Alkoxy, C 1-3 Haloalkyl, C 4-6 Haloalkyl, C 1-3 Halogenated alkoxy groups, C 4-6 Halogenated alkoxy groups.

[0030] In a preferred embodiment, R 1B R 1C Each is independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, C4 alkyl, C5 alkyl, C6 alkyl, fluorine, chlorine, cyano, amino, carboxyl, hydroxyl, methoxy, ethoxy, propoxy, isopropoxy, C4 alkoxy, C5 alkoxy, C6 alkoxy, halomethyl, haloethyl, halopropyl, haloisopropyl, C4 haloalkyl, C5 haloalkyl, C6 haloalkyl, halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, C4 haloalkoxy, C5 haloalkoxy, C6 haloalkoxy.

[0031] In a preferred embodiment, R 1B R 1C Each is independently selected from hydrogen and methyl.

[0032] In a further preferred embodiment, each R 1A Each is independently selected from methyl,

[0033] In a further preferred embodiment, each R 1A Each is independently selected from methyl,

[0034] In one preferred embodiment, R1 is selected from...

[0035] In one preferred embodiment, R1 is selected from...

[0036] In one embodiment, in the above-mentioned compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, R3 is selected from hydrogen, 3-4-membered heterocyclic alkyl, 5-12-membered heterocyclic alkyl, 4-5-membered heterocyclic group, and 6-12-membered heterocyclic group; the heterocyclic alkyl or heterocyclic group contains 1-3 heteroatoms selected from N, O, and S; any carbon atom on the heterocyclic alkyl or heterocyclic group may be oxidized; the heterocyclic alkyl or heterocyclic group is unsubstituted or oxidized by one or more R3 groups. 3A replace.

[0037] In a preferred embodiment, R3 is selected from hydrogen, 5-6-membered heterocyclic alkyl, 7-8-membered heterocyclic alkyl, 9-10-membered heterocyclic alkyl, 5-6-membered heterocyclic group, 7-8-membered heterocyclic group, and 9-10-membered heterocyclic group; the heterocyclic alkyl or heterocyclic group contains 1-3 heteroatoms selected from N, O, and S; any carbon atom on the heterocyclic alkyl or heterocyclic group can be oxidized; the heterocyclic alkyl or heterocyclic group is unsubstituted or oxidized by one or more R3 groups. 3A replace.

[0038] Alternatively, in a preferred embodiment, R3 is selected from hydrogen, 3-4 membered heterocyclic alkyl, 11-12 membered heterocyclic alkyl, 3-4 membered heterocyclic group, and 11-12 membered heterocyclic group; the heterocyclic alkyl or heterocyclic group contains 1-3 heteroatoms selected from N, O, and S; any carbon atom on the heterocyclic alkyl or heterocyclic group may be oxidized; the heterocyclic alkyl or heterocyclic group is unsubstituted or oxidized by one or more R3 groups. 3A replace.

[0039] In a further preferred embodiment, R3 is selected from hydrogen, 5-membered heterocyclic alkyl, 6-membered heterocyclic alkyl, 7-membered heterocyclic alkyl, 8-membered heterocyclic alkyl, and 6-membered heterocyclic group.

[0040] In a further preferred embodiment, R3 is selected from hydrogen, unsubstituted hydrogen, or hydrogen ions formed by one or more R3 groups. 3A Replacement

[0041] In a further preferred embodiment, R3 is selected from hydrogen, unsubstituted hydrogen, or hydrogen ions selected from one or more R3 groups. 3A Replacement

[0042] In one implementation scheme, each R 3A Each is independently selected from hydrogen and C. 1-3 Alkyl, C 4-6Alkyl, fluorine, chlorine, bromine, iodine, cyano, amino, carboxyl, hydroxyl, nitro, C 1-3 Alkoxy, C 4-6 Alkoxy, C 1-3 Haloalkyl, C 4-6 Haloalkyl, C 1-3 Halogenated alkoxy groups, C 4-6 Halogenated alkoxy groups.

[0043] In a preferred embodiment, each R 3A Each is independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, C4 alkyl, C5 alkyl, C6 alkyl, fluorine, chlorine, cyano, amino, carboxyl, hydroxyl, methoxy, ethoxy, propoxy, isopropoxy, C4 alkoxy, C5 alkoxy, C6 alkoxy, halomethyl, haloethyl, halopropyl, haloisopropyl, C4 haloalkyl, C5 haloalkyl, C6 haloalkyl, halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, C4 haloalkoxy, C5 haloalkoxy, C6 haloalkoxy.

[0044] In a more preferred embodiment, each R 3A Each is independently selected from hydrogen, methyl, fluorine, chlorine, and amino.

[0045] In one preferred embodiment, R3 is selected from hydrogen,

[0046] In one preferred embodiment, R3 is selected from hydrogen,

[0047] In one embodiment, in the above-mentioned compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, R5 is selected from hydrogen, -C(O)NR. 5B R 5C ;R 5B and R 5C Each is independently selected from hydrogen and C. 3-6 cycloalkyl, C 7-8 cycloalkyl, C 9-10 Cycloalkyl.

[0048] In a preferred embodiment, R 5B and R 5C Each is independently selected from hydrogen and C. 3-4 cycloalkyl, C 5-6 Cycloalkyl.

[0049] Or in a preferred embodiment, R 5B and R 5CEach is independently selected from hydrogen and C. 7-8 cycloalkyl, C 9-10 Cycloalkyl.

[0050] In a more preferred embodiment, R 5B and R 5C Each is independently selected from hydrogen, cyclopropane, cyclobutane, cyclopentane, and cyclohexane.

[0051] In a most preferred embodiment, each R5 is selected from hydrogen,

[0052] In one embodiment, among the above-mentioned compounds, or their prodrugs, solvates, crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, metabolites, and isotopic derivatives, compounds of formula (I) have the structure shown in formula (II):

[0053] The definitions of R1 and R3 refer to the aforementioned definitions.

[0054] In one embodiment, the compound described above, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, wherein the compound of formula (I) has the structure shown in formula (III):

[0055] Among them, R1, R 5B or R 5C The definition refers to the aforementioned definition.

[0056] In a preferred embodiment, the above-mentioned compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, wherein the compound of formula (I) has the structure shown in formula (IV-1), (IV-2), or (IV-3):

[0057] The definition of R3 references the aforementioned definition.

[0058] In one embodiment, the above-mentioned compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, is selected from the 44 specific compounds listed in Table 2.

[0059] The compounds of this application can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of this application. The chemical reactions in the specific embodiments of this application are carried out in a suitable solvent, which must be suitable for the chemical changes of this application and the reagents and materials required therefor. In order to obtain the compounds of this application, it is sometimes necessary for those skilled in the art to modify or select the synthetic steps or reaction processes based on existing embodiments.

[0060] An important consideration in planning synthetic routes in this field is selecting suitable protecting groups for reactive functional groups (such as amino groups in this application), for example, see Chem. Commun., 2019, 55, 7331-7334. All references cited in this application are incorporated herein by reference in their entirety, or may be a combination of synthetic methods known in the art and the methods described herein. The products obtained from each reaction step are obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, and chromatographic separation. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized according to literature (such as those provided by Scifinder) or purchased.

[0061] The compounds of general formulas (I), (II), (III), (IV-1), (IV-2), or (IV-3) of this invention have IC50 values ​​of less than 5 nM against MNK1 and MNK2 kinases. 50 .

[0062] Pharmaceutical composition:

[0063] The present invention also provides a pharmaceutical composition comprising a compound represented by formula (I), (II), (III), (IV-1), (IV-2) or (IV-3) above, or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, and optionally a pharmaceutical carrier and / or excipient and / or diluent.

[0064] In some embodiments, the pharmaceutical composition may also contain other medicines for treating and / or preventing and / or improving diseases associated with the biological activity of MNK1 or / and MNK2.

[0065] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredient are known, or will be obvious to those skilled in the art according to the disclosure of the present invention. As described in REMINGTON'S PHARMACEUTICAL SCIENCES, Martin, EW, ed., Mack Publishing Company, 19th ed. (1995), the method for preparing the pharmaceutical composition includes incorporating appropriate pharmaceutical excipients, carriers, diluents, etc.

[0066] Medical uses:

[0067] Another aspect of the present invention relates to the use of compounds represented by formulas (I), (II), (III), (IV-1), (IV-2 or (IV-3)) above, or their prodrugs, solvates, crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, metabolites, isotope derivatives, and the use of the above compositions in the preparation of medicaments for treating and / or preventing and / or improving diseases related to the biological activity of MNK1 or / and MNK2.

[0068] Another aspect of the invention relates to a method of treating and / or preventing and / or treating diseases associated with the biological activity of MNK1 or / and MNK2, comprising administering to an individual in need an effective amount of a compound of formula (I), (II), (III), (IV-1), (IV-2) or (IV-3) above, or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, or a combination thereof.

[0069] Another aspect of the present invention relates to compounds represented by formulas (I), (II), (III), (IV-1), (IV-2) or (IV-3) above, or their prodrugs, solvates, crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, metabolites, isotope derivatives, and compositions thereof, for the treatment and / or prevention and / or improvement of diseases associated with the biological activity of MNK1 or / and MNK2.

[0070] Another aspect of the invention relates to the use of compounds represented by formulas (I), (II), (III), (IV-1), (IV-2) or (IV-3) above, or their prodrugs, solvates, crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, metabolites, isotope derivatives, and combinations thereof, in the treatment and / or prevention and / or improvement of diseases associated with the biological activity of MNK1 or / and MNK2.

[0071] According to a preferred embodiment of the present invention, the diseases associated with the biological activity of MNK1 or / and MNK2 are selected from cancer and hyperproliferative diseases.

[0072] Technical terms of this invention:

[0073] In the following description, certain specific details are set forth to provide a thorough understanding of the various embodiments. However, those skilled in the art will understand that the invention can be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless the context otherwise requires, throughout the specification and claims, the word “comprising” and its variations, such as “including” and “containing,” should be interpreted in an open-ended sense, meaning “including but not limited to.” Furthermore, the headings provided herein are for convenience only and are not intended to define the scope or meaning of the claimed invention.

[0074] Throughout this specification, the phrase "an embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described with respect to that embodiment is included in at least one embodiment. Therefore, the phrases "in an embodiment" or "in an embodiment" appearing in different places throughout the specification do not necessarily all refer to the same embodiment. Furthermore, a particular feature, structure, or characteristic may be combined in any suitable manner in one or more embodiments. Moreover, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural indicators unless the context clearly specifies otherwise. It should also be noted that, unless the context clearly specifies otherwise, the term "or" is generally used to include the meaning of "and / or."

[0075] The term "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.

[0076] The term "prodrug" refers to a derivative of the compound of formula (I) with specific substituents discovered in this invention. These compounds may have weak or no activity on their own, but after administration, they are converted into the compound with specific substituents discovered in this invention under physiological conditions (e.g., through metabolism, solvation, or other means) to produce the corresponding biological activity in vivo.

[0077] The term "metabolite" refers to the product obtained in vivo by the metabolism of a compound of formula (I) having specific substituents, as discovered in this invention. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by experimental methods as described in this invention. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, acylation, deacylation, esterification, deesterification, enzymatic cleavage, etc. Accordingly, this invention includes metabolites of compounds, including metabolites produced by sufficiently exposing the compounds of this invention to mammals for a period of time.

[0078] The term "isotope derivative" refers to compounds of the present invention comprising at least one atom substituted with its corresponding isotopic atom. In some embodiments, the compound comprises two or more atoms substituted with their corresponding isotopic atoms. In some embodiments, the compound comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 atoms substituted with their corresponding isotopic atoms. Synthetic methods for incorporating isotopes into organic compounds are known in the art.

[0079] According to the conventions in this field, In the structural formula of this paper, the bond is used to describe the connection point between the part or substituent and the parent nucleus or main structure.

[0080] In this invention, the term "optional" means that it may or may not be replaced by the following parts.

[0081] In this invention, the term "substituted" means that an atom or group of atoms formally replaces hydrogen and is attached to another group as a "substituent". Unless otherwise stated, the term "substituted" refers to any degree of substitution where such substitution is permissible, such as monosubstituted, disubstituted, trisubstituted, tetrasubstituted, or pentasubstituted. Substituents are chosen independently, and substitution can occur at any chemically accessible position. It should be understood that substitution at a specified atom is limited by the valence of the atom. It should be understood that substitution at a specified atom produces a chemically stable molecule.

[0082] In this invention, the term "alkyl" used alone or in combination with other terms refers to a saturated hydrocarbon group that can be straight-chain or branched. The term "C 1-6"Alkyl" refers to an alkyl group having 1 to 6 carbon atoms. An alkyl group formally corresponds to an alkane where a CH bond is replaced by a junction between the alkyl group and the rest of the compound. In some embodiments, the alkyl group contains 1-6 carbon atoms, 1-4 carbon atoms, 1-3 carbon atoms, or 1-2 carbon atoms. Examples of alkyl groups include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, primary butyl, tert-butyl, and isobutyl; higher homologues such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, and 1,2,2-trimethylpropyl.

[0083] In this invention, the term "halogenated" or "halogen" used alone or in combination with other terms refers to F, Cl, Br, and I. In some embodiments, the term "halogenated" refers to a halogen atom selected from F, Cl, or Br.

[0084] In this invention, the term "halogenated alkyl" used alone or in combination with other terms refers to an alkyl group substituted with one or more halogens, wherein the terms "halogen" and "alkyl" are as defined above.

[0085] In this invention, the term "heteroatoms" used alone or in combination with other terms includes S, O, and N.

[0086] In this invention, the term "alkoxy" used alone or in combination with other terms refers to the formula -OR a The group, wherein R a The alkyl group is as defined above. Unless otherwise specifically indicated in this specification, the alkoxy group may optionally be substituted as described below, for example, by oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc. In some embodiments, the alkoxy group is optionally substituted by halogen, -CN, -OH, -OMe, -NH2, or -NO2. In some embodiments, the alkoxy group is optionally substituted by halogen, -CN, -OH, or -OMe. In some embodiments, the alkoxy group is optionally substituted by halogen.

[0087] In this invention, the term "haloalkoxy" used alone or in combination with other terms refers to an alkoxy group substituted with one or more halogens, wherein the terms "halogen" and "alkoxy" are as defined above.

[0088] In this invention, the term "heteroaryl," used alone or in combination with other terms, refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from S, O, and N, wherein the heteroatom can be on any ring or simultaneously on multiple (e.g., two or three) rings. In some embodiments, the heteroaryl ring has 1, 2, 3, or 4 heteroatom ring members independently selected from S, O, and N. In some embodiments, any cyclic N in the heteroaryl moiety can be an N-oxide. In some embodiments, the heteroaryl has 4 to 12 ring atoms, including a carbon atom and 1, 2, 3, or 4 heteroatom ring members independently selected from S, O, and N. In some embodiments, the heteroaryl has 4 to 10 ring atoms, including a carbon atom and 1, 2, 3, or 4 heteroatom ring members independently selected from S, O, and N. In some embodiments, the heteroaryl has 5 to 6 ring atoms and 1 or 2 heteroatom ring members independently selected from S, O, and N. The cyclic carbon atom and heteroatom of the heteroaryl group can optionally be oxidized to form an oxo or sulfide ion group or other oxidized bond (e.g., C(O), S(O), C(S) or S(O)2, N-oxide, etc.), or the nitrogen atom can be quaternized. The heteroaryl group can be linked via a cyclic carbon atom or a cyclic heteroatom. In some embodiments, the heteroaryl group is a five- or six-membered heteroaryl ring. In other embodiments, the heteroaryl group is an eight-, nine-, or ten-membered fused bicyclic heteroaryl ring. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyridazinyl, pyrroleyl, pyrazolyl, azoleyl, oxazolyl, thiazolyl, imidazoleyl, furanyl, thiophenyl, quinolinyl, isoquinolinyl, naphthinyl (including 1,2-naphthidine, 1,3-naphthidine, 1,4-naphthidine, 1,5-naphthidine, 1,6-naphthidine, 1,7-naphthidine, 1,8-naphthidine, 2,3-naphthidine and 2,6-naphthidine), indoleyl, benzothiophenyl, benzofuranyl, benzoisoxazolyl, imidazole[1,2-b]thiazolyl, purineyl, etc.

[0089] In this invention, the term "heterocyclic group," used alone or in combination with other terms, refers to substituted and unsubstituted 4-8 membered monocyclic or bicyclic groups, 8-10 membered bicyclic or tricyclic groups, and 10-12 membered tricyclic or polycyclic groups, wherein at least one ring has at least one heteroatom (O, S, or N), and the heteroatom-containing ring preferably has one, two, or three heteroatoms selected from O, S, and N. Each heteroatom-containing ring in the group may contain one or two oxygen or sulfur atoms and / or one to four nitrogen atoms, with the limitation that the total number of heteroatoms in each ring is four or less, and a further limitation that the ring contains at least one carbon atom. In some preferred embodiments, the heteroatom refers only to N or O, and its total number does not exceed three, preferably containing only one to two heteroatoms. Carbon and sulfur atoms may optionally be oxidized, nitrogen atoms may optionally be quaternized, and ring atoms on the heterocycle may optionally be substituted with =O (oxo) when the valence allows. (For example: Fused rings completing bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or fully unsaturated, aromatic or non-aromatic. Heterocyclic groups may be attached to any available nitrogen or carbon atom. The term "heterocycle" as used herein does not include fully aromatic rings or heterocyclic alkyl groups.

[0090] In this invention, the term "cycloalkyl" used alone or in combination with other terms refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic), where polycyclic (e.g., bicyclic or tricyclic) includes fused rings, spirocyclic, and bridged rings, including cyclized alkyl groups. The term "C"... 3-12 "Cycloalkyl" or "C" 3-10 "Cycloalkyl" refers to a cycloalkyl group having 3-12 or 3-10 ring member carbon atoms, respectively. Cycloalkyl groups can include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) groups and spirocyclic groups. Cycloalkyl groups can have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 cyclic carbons (C atoms). 3-12 In some embodiments, the cycloalkyl group has 3-12 ring members, 3-10 ring members, 3-8 ring members, 3-6 ring members, 3-5 ring members, or 3-4 ring members. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is monocyclic or bicyclic. In some embodiments, the cycloalkyl group is C10. 3-8 Monocyclic cycloalkyl group. The cycloforming carbon atom of the cycloalkyl group may optionally be oxidized to form an oxo or sulfide ion group. In some embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. In some embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0091] In this invention, unless otherwise specified, the term "heterocyclic alkyl" used alone or in combination with other terms refers to a non-aromatic ring system (monocyclic, bicyclic, or polycyclic), where polycyclic (e.g., bicyclic or tricyclic) includes fused rings, spirocyclic, and bridged rings, having at least one heteroatom ring member independently selected from N, S, and O, and having 4-12, 4-10, or 4-8 ring members. The heteroatom can be on any ring or simultaneously on multiple (e.g., two or three) rings. The term "heterocyclic alkyl" includes monocyclic 4-, 5-, 6-, 7-, and 8-membered heterocyclic alkyl groups. In some embodiments, a heterocyclic alkyl group is a monocyclic group having 1, 2, or 3 heteroatoms independently selected from N, S, and O. In some embodiments, a heterocyclic alkyl group is a spirocyclic, bridged, or fused ring having 1, 2, or 3 heteroatoms independently selected from N, S, and O. The cyclic carbon atoms and heteroatoms of heterocyclic alkyl groups may optionally be oxidized to form oxo or thio groups or other oxidized bonds (e.g., C(O), S(O), C(S) or S(O)2, N-oxides, etc.), or the nitrogen atom may be quaternized. Heterocyclic alkyl groups may be linked via cyclic carbon atoms or cyclic heteroatoms.

[0092] In this invention, the term "aryl" used alone or in combination with other terms refers to an aromatic hydrocarbon group, which can be monocyclic or polycyclic (e.g., having two fused rings). The term "C 4-12 "Aryl" refers to an aryl group having 4 to 12 ring carbon atoms. The term "C" is used in this context. 4-10 "Aryl" refers to an aryl group having 4 to 10 ring carbon atoms. The term "C" is used in this context. 6-10 "Aryl" refers to an aryl group having 6 to 10 cyclic carbon atoms. Aryl groups include, for example, phenyl, naphthyl, indenyl, and indene. In some embodiments, the aryl group has 6 carbon atoms. In some embodiments, the aryl group has 10 carbon atoms. In some embodiments, the aryl group is phenyl. In some embodiments, the aryl group is naphthyl.

[0093] In this invention, the term "therapeutic and / or preventive and / or ameliorative effective amount" used alone or in combination with other terms refers to the amount of a compound administered as a single dose or as part of a series of doses to mammals and / or subjects that effectively produces the desired therapeutic and / or preventive and / or ameliorative effect.

[0094] The beneficial effects of this invention are:

[0095] Firstly, the compounds of the present invention exhibit IC50 values ​​below 5 nM against both MNK1 and MNK2 kinases. 50 Therefore, the compounds and pharmaceutical compositions provided by the present invention exhibit unexpectedly excellent MNK1 and / or MNK2 inhibitory effects, and can be used as medicines for the prevention and / or improvement and / or treatment of diseases related to these effects.

[0096] Secondly, after oral administration to mice, the compound of the present invention exhibits good exposure in vivo, with its plasma exposure (AUC) and peak concentration (Cmax) being significantly higher than those of the control molecule O1, which has a similar structure, and is expected to bring better potential anti-tumor activity.

[0097] Thirdly, after oral administration to mice, the compounds of the present invention are mainly distributed in solid tissues such as the liver, lungs, and kidneys. The ratio of drug concentration in tissues to plasma is significantly greater than 1, and their tissue distribution is significantly higher than that of the control molecule O1, which has a similar structure. This indicates that compared with the control molecule O1, the compounds of the present invention have a stronger affinity for tissues and have the potential to exert a stronger therapeutic effect in these tissues.

[0098] Fourthly, after oral administration to mice, the compound of this invention exhibited tumor-targeting properties, with a drug concentration ratio of 5.2 in tumor tissue to plasma, indicating a high accumulation of the drug at the tumor site, providing a strong material basis for enhancing anti-tumor efficacy. Furthermore, in preliminary pharmacodynamic tests, oral administration of the compound of this invention to mice significantly reduced tumor volume, further demonstrating that the compound can target tumors and exert a significant effect, a result that was unexpected. Detailed Implementation Plan

[0099] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Various other modifications, substitutions, or alterations can be made based on ordinary technical knowledge and common practice in the art, without departing from the basic technical concept of this invention described above.

[0100] The present invention will be described in detail below with reference to examples, but this does not imply any adverse limitation on the invention. The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention, and these should also be considered within the scope of protection of the present invention.

[0101] The structure of the compound was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker ASCENA-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻⁶. -6(ppm) is given as the unit.

[0102] Reaction monitoring and MS determination were performed using a Thermofisher ESQ (ESI) mass spectrometer.

[0103] The HPLC determination was performed using a Thermo Fisher U3000 DAD high-performance liquid chromatograph (GL Sciences ODS-HL HP 3μm 3.0*100mm column).

[0104] Thin-layer chromatography (TLC) used Qingdao Ocean GF254 silica gel plates. The silica gel plates used in TLC had a diameter of 0.15–0.2 mm, while the purified products were separated using high-performance thin-layer chromatography (HPLC) preparative plates with a diameter of 0.9–1.0 mm. Column chromatography used Qingdao Ocean 200–300 mesh silica gel as the carrier. The developing solvent systems were A: dichloromethane and methanol; and B: petroleum ether and ethyl acetate. The volume ratio of the solvents was adjusted according to the polarity of the compounds. For medium-pressure preparative liquid chromatography (PLC) purification, a Biotage Isera One preparative PL was used.

[0105] In the following examples, unless otherwise specified, all reaction materials can be purchased from manufacturers such as Shanghai Shaoyuan Reagent Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., and Shanghai Bide Pharmaceutical Technology Co., Ltd.

[0106] Table 1: Chinese / English / Abbreviated Definitions

[0107] This application prepared 44 compounds as shown in Table 2 below:

[0108] Table 2: Compound Structures in Examples

[0109] The preparation process of representative compounds is described in detail below, while the other compounds were prepared by similar synthetic methods.

[0110] Example 3: Preparation of 3-[1-methyl-7-(3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (3)

[0111] Step 1: Preparation of tert-butyl 3-(2-fluoro-3-nitrophenyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0112] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (2.2 g, 10.00 mmol), tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate hydrochloride (2.5 g, 10.00 mmol), Cs₂CO₃ (5.3 g, 16.26 mmol), XPhos Pd G₃ (0.9 g, 1.00 mmol), and dioxane solvent (50 mL) were added. The reaction mixture was stirred overnight at 110 °C under N₂. The reaction mixture was filtered, the filtrate was evaporated to dryness, the reaction was quenched with water, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and washed with saturated NaHCO₃ (aq) and saturated brine. The mixture was dried over anhydrous Na₂SO₄, filtered, concentrated, and the crude product was purified by column chromatography (EA:PE = 1:5) to give 2.5 g of a brown oil. The product is tert-butyl 3-(2-fluoro-3-nitrophenyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.5 g, 7.11 mmol, 71.1%). m / z = 296.2 [M+H-56] + .

[0113] Step 2: Preparation of 3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester

[0114] In a 250 mL round-bottom flask, tert-butyl 3-(2-fluoro-3-nitrophenyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.5 g, 7.11 mmol), methylamine hydrochloride (0.7 g, 10.67 mmol), potassium carbonate (2.9 g, 21.34 mmol), and acetonitrile (50 mL) were added. The mixture was stirred at 70 °C for 3 h, and the reaction was monitored by LC-MS until it was essentially complete. The reaction solution was cooled, filtered, and the filter cake was washed three times with a small amount of acetonitrile. The filtrate and washings were collected, evaporated to dryness, and the reaction solution was added dropwise to water, precipitating a yellow solid. The solid was filtered, the filter cake was washed with water, and dried to obtain 2.3 g of a reddish-brown solid. That is, 3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester (2.3 g, 6.35 mmol, 89.2%). m / z = 363.2 [M+H] + .

[0115] Step 3: Preparation of 3-[3-amino-2-(methylamino)phenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester

[0116] In a 250 mL round-bottom flask, 2.5 g (6.90 mmol) of 3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester, ethanol (100 mL), and water (10 mL) were added. Sodium hydrosulfite (28.8 g, 165.56 mmol) was added in three portions at room temperature, and the mixture was stirred for 30 minutes at room temperature until the reaction solution became colorless. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 90 / 10) to obtain 600 mg of a yellow solid. That is, 3-[3-amino-2-(methylamino)phenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester (600 mg, 1.81 mmol, 26.2%). m / z = 333.4 [M+H] + .

[0117] Step 4: Preparation of 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine]

[0118] In a 100 mL round-bottom flask, add the reactants 3-[3-amino-2-(methylamino)phenyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester (150 mg, 0.45 mmol), 3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-carboxylic acid (150 mg, 0.52 mmol), methylimidazolium (120 mg, 1.46 mmol), and acetonitrile (20 mL) as solvent. Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (170 mg, 0.61 mmol) and react overnight at room temperature. The next day, add acetic acid (2 mL) to the reaction mixture and stir at 80 °C for 2 hours. The reaction solution was added dropwise to water (50 mL), and a large amount of yellow solid precipitated. After filtration, the filter cake was sonicated for 5 minutes with methanol / dichloromethane (2 mL / 5 mL), and then slurried with methanol to obtain 200 mg of yellow solid, namely 3-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzo[d]imidazol-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid-2-methylpropyl-2-yl ester (200 mg, 0.34 mmol, 75.7%).

[0119] Step 5: Preparation of 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (3)

[0120] In a 50 mL single-necked flask, add the reactant (2S)-4-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1H-pyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-2-methylpiperazine-1-carboxylic acid-2-methylpropyl-2-yl ester (20 mg, 0.04 mmol), hydrochloric acid (0.5 mL, 64.67 mmol), and solvent MeOH (2 mL). The reaction solution was stirred at room temperature for 1 hour, concentrated, and the pH of the solution was adjusted to weakly acidic with sodium bicarbonate. The solution was then filtered directly, evaporated to dryness, and the crude product was purified by high-performance liquid chromatography (HPLC) to obtain 9 mg of a yellow solid, namely the product 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (12) (9 mg, 0.02 mmol, 52.9%). m / z = 461.2 [M+H] + HPLC: 96.51%.

[0121] 1 H NMR (400MHz, DMSO) δ8.51(s,1H),8.39(s,1H),8.08(s,1H),7.88(s,2H),7.60(dd,J=7.0,1.8Hz,1H),7.36-7.22(m,2H),4.62(s,3H),4.42-4.4 4(m,1H),4.12(s,2H),3.99(d,J=11.3Hz,2H),3.50(m,2H),3.42-3.38( m,2H),3.19(d,J=10.5Hz,2H),2.27(d,J=7.7Hz,2H),2.05-2.01(m,6H).

[0122] Example 4: Preparation of 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (4)

[0123] Add the reactants 3-[1-methyl-7-(3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (30 mg, 0.06 mmol), potassium carbonate (15 mg, 0.11 mmol), and DMF (4 mL) to a 10 mL microwave-safe tube. Then add 0.1 mL of a DMF solution containing 10 mg, 0.11 mmol of iodomethane. Stir the reaction mixture at room temperature for about 12 h. Monitor the reaction until it is complete by LC-MS the following day. The reaction solution was directly evaporated to dryness, and then purified by column chromatography to obtain 18 mg of a yellow solid, namely 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (18 mg, 0.03 mmol, 56.3%). m / z = 500.30 [M+H] + HPLC: 96.57%.

[0124] 1 H NMR (400MHz, DMSO)8.52(s,1H),8.39(s,1H),8.09(s,1H),7.89(s,2H),7.62(d,J=8.6Hz,1H),7.35-7.18(m,2H),4.63(s,3H),4.54-4.4 0(m,1H),4.07(s,2H),3.99(d,J=11.3Hz,2H),3.52-3.47(m,3H),3.44-3.38(m,3H),2.83(s,3H),2.36-2.32(m,4H),2.08-1.97(m,4H).

[0125] Example 6: Preparation of 5-{5-amino-6-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (6)

[0126] Step 1: Preparation of 3-(2-fluoro-3-nitrophenyl)-8-methyl-3,8-diazabicyclo[3.2.1]octane

[0127] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (6 g, 27.27 mmol), 8-methyl-3,8-diazabicyclo[3.2.1]octane dihydrochloride (5 g, 25.11 mmol), cesium carbonate (13.3 g, 40.91 mmol), XPhos Pd G4 (2.3 g, 2.73 mmol), and dioxane solvent (500 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, the reaction was quenched with water, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and washed with saturated NaHCO3 (aq) and saturated brine. The mixture was dried over anhydrous Na2SO4, filtered, concentrated, and the crude product was purified by column chromatography (DCM / MeOH = 92 / 8) to give 2.2 g of a brown oil. This yielded 3-(2-fluoro-3-nitrophenyl)-8-methyl-3,8-diazabicyclo[3.2.1]octane (2.2 g, 6.78 mmol, 25.0%). m / z = 266.1 [M+H] + .

[0128] Step 2: Preparation of 8-methyl-3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane

[0129] In a 250 mL round-bottom flask, 2.2 g (6.78 mmol) of 3-(2-fluoro-3-nitrophenyl)-8-methyl-3,8-diazabicyclo[3.2.1]octane, methylamine hydrochloride (1.5 g, 22.62 mmol), potassium carbonate (7.3 g, 52.77 mmol), and 50 mL of acetonitrile were added. The mixture was stirred overnight at 80°C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain 4.0 g of crude brown solid product, namely 8-methyl-3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane (2.0 g, 3.62 mmol, 47.6%). m / z = 277.15 [M+H] + .

[0130] Step 3: Preparation of 2-(methylamino)-3-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)aniline

[0131] In a 250 mL round-bottom flask, 8-methyl-3-[2-(methylamino)-3-nitrophenyl]-3,8-diazabicyclo[3.2.1]octane (2.0 g, 7.24 mmol), methanol (100 mL), and water (15 mL) were added. Sodium hydrosulfite (25.2 g, 144.77 mmol) was added in three portions at room temperature, and the mixture was stirred for 3 h at room temperature until the reaction solution became colorless. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 90 / 10) to obtain 1.2 g of a yellow solid, namely 2-(methylamino)-3-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)aniline (1.2 g, 4.87 mmol, 67.3%). m / z = 247.2 [M+H]. + .

[0132] Step 4: Preparation of 5-bromo-3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-amine

[0133] In a 100 mL round-bottom flask, add the reactants 2-(methylamino)-3-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)aniline (1.2 g, 4.87 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (1.1 g, 4.87 mmol), methylimidazole (0.8 g, 9.74 mmol), and solvent acetonitrile (50 mL). Add Tcfh (1.8 g, 6.33 mmol) and react overnight at room temperature. The next day, add acetic acid (2 mL) to the reaction mixture and stir at 80 °C for 2 hours. The reaction mixture was added dropwise to saturated sodium bicarbonate solution (50 mL), extracted three times with ethyl acetate, and the organic phases were combined, evaporated to dryness, and purified by column chromatography (DCM / MeOH = 95 / 5) to give 660 mg of a yellow solid, namely 5-bromo-3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-amine (660 mg, 1.54 mmol, 31.4%). m / z = 428.1 / 430.1 [M+H] + .

[0134] Step 5: Preparation of 5-bromo-2,N,N-trimethylpyrazole-3-carboxamide

[0135] In a 100 mL round-bottom flask, 5-bromo-2-methylpyrazole-3-carboxylic acid (1.1 g, 5.37 mmol), dimethylamine hydrochloride (0.6 g, 7.36 mmol), methylimidazole (1.2 g, 14.62 mmol), and acetonitrile (50 mL) were added as solvent. TCFH N,N,N',N'-tetramethylchloroformamidin hexafluorophosphate (2.0 g, 7.13 mmol) was added, and the mixture was reacted at room temperature for 2 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted three times with 30 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give 1.0 g of a yellow solid, namely 5-bromo-2,N,N-trimethylpyrazole-3-carboxamide (1.0 g, 4.31 mmol, 80.3%). m / z = 231.9 / 233.9 [M+H] + .

[0136] Step 6: Preparation of 5-{5-amino-6-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (6)

[0137] The following ingredients were added: 5-bromo-3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-amine (150 mg, 0.38 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1,3,2-dioxaborane (150 mg, 0.60 mmol), Pd(dppf)Cl2 (40 mg, 0.05 mmol), and KOAc (70 mg, 0.71 mmol). Add 170 mg (0.73 mmol) to dioxane (10 mL), and under N2 protection, stir and react in a microwave at 120 °C for 2 hours. Transfer the reaction solution to a 50 mL reaction flask, and add 5-bromo-2,N,N-trimethylpyrazole-3-carboxamide (170 mg, 0.73 mmol), Cs2CO3 (200 mg, 0.61 mmol), and Pd(dppf)Cl2 (50 mg, 0.07 mmol) to the reaction flask. Add dioxane-water (6 mL), and under N2 protection, stir and react in an oil bath at 110 °C for 1 hour. The product was diluted with water, extracted with EA, and the organic phase was concentrated. The crude product was purified by HPLC to obtain 26 mg of a yellow solid, namely the product 5-{5-amino-6-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (26 mg, 0.05 mmol, 13.6%). m / z = 501.3 [M+H] + .

[0138] HPLC: 99.61%. 1 H NMR (400MHz, DMSO) δ8.64(s,1H),8.19(s,1H),8.08(s,2H),7.51(dd,J=5.3,3.7Hz,1H),7.27-7.16(m,2H),7.00(s,1H),4.62(s,3 H),3.90(s,3H),3.25(s,2H),3.11(d,J=10.3Hz,2H),3.08(s,3H),3.03(s,3H),3.00-2.96(m,2H),2.30(s,3H),2.03-1.97(m,4H).

[0139] Example 9: Preparation of 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-(1-methylpyrazol-3-yl)pyrazin-2-amine (9)

[0140] In a 100 mL round-bottom flask, add the reactants 5-bromo-3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]pyrazin-2-amine (155 mg, 0.36 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)pyrazole (100 mg, 0.48 mmol), PdCl2(dppf) (50 mg, 0.07 mmol), Cs2CO3 (200 mg, 0.61 mmol), water (3 mL), and dioxane solvent (15 mL). Stir the reaction mixture overnight in N2 at 110 °C. The reaction solution was evaporated to dryness. The crude product was purified by column chromatography (DCM / MeOH = 95 / 5) to obtain 65 mg of a yellow solid. This crude product was then sonicated with ethyl acetate (2 mL) for 5 min, filtered, and the filter cake was washed with a small amount of ethyl acetate. The filter cake was collected and dried under reduced pressure to obtain 35 mg of a yellow solid, namely 3-[1-methyl-7-(8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)benzo[d]imidazol-2-yl]-5-(1-methylpyrazol-4-yl)pyrazin-2-amine (35 mg, 0.08 mmol, 21.3%). m / z 430.3 [M+H] + .

[0141] HPLC: 94.56%. 1H NMR (400MHz, DMSO) δ8.49(s,1H),8.27(s,1H),8.03(s,1H),7.90(s,2H),7.58(d,J=5.2Hz,1H),7.2 8(d,J=5.5Hz,2H),4.63(s,3H),3.93-3.80(m,5H),3.17(d,J=8.9Hz,2H),2.67(s,3H),2.24(s,4H).

[0142] Example 11: Preparation of 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazole-3-carboxamide (11)

[0143] Step 1: Preparation of {[1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0144] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (6.0 g, 27.27 mmol), (hexahydropyridin-3-ylamino)methane-2-methylpropyl-2-yl ester (6.0 g, 29.96 mmol), cesium carbonate (17 g, 52.18 mmol), XPhos Pd G4 (2.4 g, 2.79 mmol), and 1,4-dioxane solvent (200 mL) were added. The reaction mixture was stirred overnight at 110 °C under nitrogen protection. The reaction mixture was filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 30 / 70) to give 6.0 g of a brown oily substance. This yielded {[1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (6 g, 17.68 mmol, 64.5%). m / z 296.2 [M+H-56] + .

[0145] Step 2: Preparation of ({1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester)

[0146] In a 250 mL round-bottom flask, the reactants {[1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (6.0 g, 17.68 mmol), methylamine hydrochloride (1.8 g, 26.67 mmol), potassium carbonate (6 g, 43.42 mmol), and solvent acetonitrile (100 mL) were added and stirred overnight at 70 °C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 30 / 70) to obtain 6.0 g of crude brown oily product. That is, ({1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (6.0 g, 17.12 mmol, 96.8%). m / z 351.1 [M+H] + .

[0147] Step 3: Preparation of ({1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester

[0148] In a 500 mL round-bottom flask, the reactant ({1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (6.0 g, 17.12 mmol), methanol (200 mL), and water (40 mL) were added. Sodium hydrosulfite (50 g, 287.19 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 95 / 5) to obtain 4.0 g of a yellow oily substance. This is ({1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (4.0 g, 12.48 mmol, 72.7%). m / z 321.2 [M+H] + .

[0149] Step 4: Preparation of ({1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester)

[0150] In a 250 mL round-bottom flask, add the reactants ({1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (4 g, 12.48 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (3.2 g, 14.68 mmol), methylimidazolium (3.0 g, 36.54 mmol), and acetonitrile (100 mL) as solvent. Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (4.2 g, 14.97 mmol) and react at room temperature for 2 hours. Add acetic acid (2 mL) to the reaction mixture, heat to 100 °C, and stir for 2 hours. The reaction mixture was rotated until acetonitrile was removed. A saturated sodium bicarbonate aqueous solution (4 mL) was added to the crude product. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined and evaporated to dryness. The crude product was purified by column chromatography (dichloromethane / methanol = 97 / 3) to give 5.7 g of a yellow solid, namely ({1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (5.7 g, 11.35 mmol, 90.9%). m / z 502.0 / 504.0 [M+H] + .

[0151] Step 5: Preparation of 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2-methylpyrazol-3-carboxylic acid

[0152] In a 150 mL round-bottom flask, add the reactants ({1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]hexahydropyridin-3-yl}amino)methane-2-methylpropyl-2-yl ester (2.5 g, 4.98 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)pyrazole-3-carboxylate (1.6 g, 6.01 mmol), Cs2CO3 (3.2 g, 9.82 mmol), PdCl2 (dppf) (0.3 g, 0.41 mmol), water (10 mL), and solvent 1,4-dioxane (50 mL). The reaction mixture is stirred overnight at 100 °C under nitrogen protection. The reaction mixture was evaporated to dryness, and the pH was adjusted with 6N hydrochloric acid. Methanol and water (10 mL) were added, and the mixture was sonicated for 15 min. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 80 / 20) to obtain 2.0 g of a yellow solid, namely 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2-methylpyrazol-3-carboxylic acid (2.0 g, 4.47 mmol, 89.7%). m / z 448.1 [M+H] + .

[0153] Step 6: Preparation of 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (11)

[0154] In a 100 mL round-bottom flask, add the reactants 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2-methylpyrazol-3-carboxylic acid (200 mg, 0.45 mmol), dimethylamine hydrochloride (40 mg, 0.59 mmol), methylimidazolium (200 mg, 2.44 mmol), and solvent acetonitrile (15 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (160 mg, 0.57 mmol) and react at room temperature for 12 hours. Add water (20 ml) to the reaction mixture, extract three times with 30 ml of dichloromethane each time, combine the organic phases, dry over anhydrous sodium sulfate, filter, and evaporate to dryness to obtain 200 mg of a colorless oil. The crude product is purified by column chromatography (dichloromethane / methanol = 85 / 15) to obtain 66 mg of a yellow solid, namely 5-{5-amino-6-[7-(3-aminohexahydropyridin-1-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (66 mg, 0.13 mmol, 29.6%). m / z 475.2 [M+H] + HPLC: 95.71%.

[0155] 1 H NMR (400MHz, DMSO) δ8.66 (s, 1H), 8.27 (s, 2H), 7.53 (d, J = 8.0Hz, 1H), 7.23 (t ,J=7.9Hz,1H),7.03(t,J=3.7Hz,2H),4.57(s,3H),3.91(s,3H),3.71(s,2H) ,3.59(d,J=10.3Hz,1H),3.07(d,J=25.5Hz,6H),2.66(dd,J=23.6,13.0Hz,2 H),2.15(d,J=10.2Hz,1H),2.02-1.72(m,2H),1.45(td,J=12.2,5.1Hz,1H).

[0156] Example 18: Preparation of 3-[7-(3-amino-5-chlorohexahydropyridin-1-yl)-1-methylbenzi[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (18)

[0157] Step 1: Preparation of tert-butyl 3-(2-fluoro-3-nitrophenyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate

[0158] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (2.8 g, 12.73 mmol), 3,6-diazabicyclo[3.1.1]heptane-3-carboxylic acid-2-methylpropyl-2-yl ester (2.6 g, 13.11 mmol), Cs₂CO₃ (7.3 g, 22.40 mmol), XPhos Pd G₃ (1.1 g, 1.27 mmol), and 1,4-dioxane (100 mL) solvent were added. The reaction mixture was stirred overnight at 110 °C under N₂. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (ethyl acetate / petroleum ether = 0 / 100–50 / 50, 15 min) to obtain 3.5 g of a brown oily substance. This yielded tert-butyl 3-(2-fluoro-3-nitrophenyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (3.5 g, 10.37 mmol, 81.5%). m / z 282.2 [M+H-56] + .

[0159] Step 2: Preparation of tert-butyl 3-[2-(methylamino)-3-nitrophenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate

[0160] In a 250 mL round-bottom flask, the reactants 3-(2-fluoro-3-nitrophenyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester (3.5 g, 10.37 mmol), methylamine hydrochloride (1.1 g, 15.56 mmol), potassium carbonate (4.3 g, 31.12 mmol), and solvent acetonitrile (50 mL) were added. The mixture was stirred at 70 °C for 3 h. The reaction solution was cooled, filtered, and the filter cake was washed three times with a small amount of acetonitrile. The filtrate and washings were collected and evaporated to dryness to obtain 3.4 g of a reddish-brown solid, namely 3-[2-(methylamino)-3-nitrophenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester (3.4 g, 9.76 mmol, 94.1%). m / z 349.2 [M+H] + .

[0161] Step 3: Preparation of 3-[3-amino-2-(methylamino)phenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester

[0162] In a 250 mL round-bottom flask, the reactant 3-[2-(methylamino)-3-nitrophenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester (3.5 g, 10.05 mmol), ethanol (100 mL), and water (20 mL) were added. Sodium hydrosulfite (42.0 g, 241.10 mmol) was added in three portions at room temperature, and the mixture was stirred for 30 minutes at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 40 / 60) to obtain 800 mg of a white solid, namely 3-[3-amino-2-(methylamino)phenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester (800 mg, 2.51 mmol, 25.0%). m / z 319.21 [M+H] + .

[0163] Step 4: Preparation of 3-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzo[d]imidazol-4-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester)

[0164] In a 100 mL round-bottom flask, add the reactants 3-[3-amino-2-(methylamino)phenyl]-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester (150 mg, 0.45 mmol), 3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-carboxylic acid (300 mg, 1.04 mmol), methylimidazolium (123.8 mg, 1.51 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (200 mg, 0.71 mmol) and react overnight at room temperature. The next day, acetic acid (2 mL) was added to the reaction solution, which was then heated to 100°C and stirred for 2 hours. The reaction solution was then added dropwise to water (50 mL), resulting in the precipitation of a large amount of yellow solid. The mixture was filtered, and the filter cake was sonicated with methanol / dichloromethane (2 mL / 5 mL) for 5 minutes. The resulting slurry was then beaten with methanol to obtain 250 mg of the yellow solid, namely 3-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzo[d]imidazol-4-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester (250 mg, 0.44 mmol, 87.0%). m / z 572.30 [M+H] + .

[0165] Step 5: Preparation of 3-[7-(3-amino-5-chlorohexahydropyridin-1-yl)-1-methylbenzi[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (18)

[0166] In a 50 mL round-bottom flask, add the reactants 3-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzo[d]imidazol-4-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid-2-methylpropyl-2-yl ester (250 mg, 0.44 mmol), MeOH (10 mL), and concentrated hydrochloric acid (36%-38%) (2 mL, 64.66 mmol). Stir the reaction mixture at room temperature for 1 hour. The reaction solution was directly evaporated to dryness, dissolved in ethyl acetate, and the filtrate was evaporated to dryness. The solution was then purified by column chromatography to obtain 3-[7-(3-amino-5-chlorohexahydropyridin-1-yl)-1-methylbenzi[d]imidazol-2-yl]-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (100 mg, 0.19 mmol, 44.0%). m / z 508.2 [M+H] + .

[0167] HPLC: 98.31%. 1 H NMR(400MHz,DMSO)δ8.50(s,1H),8.37(s,1H),8.06(s,1H),7.94(s,2H),7.50(d, J=8.0Hz,1H),7.20(t,J=7.8Hz,1H),6.99(d,J=7.6Hz,1H),4.81(s,1H),4.64(s, 3H),4.49(dd,J=14.8,8.6Hz,1H),3.99(dd,J=8.4,2.9Hz,2H),3.62-3.41(m,4H) ,3.31-3.24(m,2H),2.37-2.09(m,2H),2.10-1.94(m,4H),1.66(t,J=11.1Hz,1H).

[0168] Example 26: Preparation of 5-(5-amino-6-{7-[(3S)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (26)

[0169] Step 1: Preparation of {[(3S)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester]

[0170] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (4.4 g, 20.00 mmol), {[(3S)-hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.0 g, 20.00 mmol), cesium carbonate (13.0 g, 39.90 mmol), XPhos Pd G4 (1.7 g, 2.00 mmol), and solvent 1,4-dioxane (100 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (dichloromethane / methanol = 92 / 8) to give 4.5 g of a brown oily substance. This yielded {[(3S)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.5 g, 13.26 mmol, 66.2%). m / z 340.08 [M+H] + .

[0171] Step 2: Preparation of {[(3S)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0172] In a 250 mL round-bottom flask, the reactants {[(3S)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.5 g, 13.26 mmol), methylamine hydrochloride (1.5 g, 22.22 mmol), potassium carbonate (7.7 g, 55.72 mmol), and solvent acetonitrile (100 mL) were added and stirred overnight at 70°C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 30 / 70) to obtain 4.0 g of crude brown oily product. That is, {[(3S)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.0 g, 11.42 mmol, 87.0%). m / z 351.1 [M+H] + .

[0173] Step 3: Preparation of {[(3S)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0174] In a 500 mL round-bottom flask, the reactant {[(3S)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.0 g, 11.42 mmol), methanol (100 mL), and water (30 mL) were added. Sodium hydrosulfite (20 g, 114.88 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 2.4 g of a yellow oily substance, namely {[(3S)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.4 g, 7.49 mmol, 65.6%). m / z 321.2 [M+H] + .

[0175] Step 4: Preparation of {[(3S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0176] In a 100 mL round-bottom flask, the reactants {[(3S)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.4 g, 7.49 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (1.9 g, 8.72 mmol), methylimidazolium (1.5 g, 18.27 mmol), and solvent acetonitrile (100 mL) were added. TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (2.7 g, 9.62 mmol) was added, and the reaction was carried out at room temperature for 12 hours. Then, acetic acid (2 mL) was added to the reaction mixture, and the temperature was raised to 100 °C and stirred for 2 hours. The reaction mixture was rotated until acetonitrile was removed. A saturated sodium bicarbonate aqueous solution (4 mL) was added to the crude product. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined and evaporated to dryness. The crude product was purified by column chromatography (dichloromethane / methanol = 97 / 3) to give 1.5 g of a yellow solid, namely {[(3S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.4 g, 4.78 mmol, 63.8%). m / z 502.0 / 504.0 [M+H] + .

[0177] Step 5: Preparation of methyl 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate

[0178] In a 100 mL round-bottom flask, add the reactants {[(3S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (800 mg, 1.59 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)pyrazole-3-carboxylate (500 mg, 1.88 mmol), cesium carbonate (800 mg, 2.46 mmol), PdCl2 (dppf) (100 mg, 0.14 mmol), water (4 mL), and solvent 1,4-dioxane (20 mL). Stir the reaction mixture in N2 at 100 °C for about 3 hours. The reaction solution was evaporated to dryness, and the pH was adjusted with 3N dilute hydrochloric acid. Methanol and water (4 mL) were added, and the mixture was sonicated for 15 min. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to obtain 600 mg of a yellow solid, namely methyl 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate (600 mg, 1.07 mmol, 67.1%). m / z 562.14 [M+H] + .

[0179] Step 6: Preparation of 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0180] In a 25 mL round-bottom flask, methyl 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate (300 mg, 0.53 mmol) and solvents MeOH (15 mL), water (3 mL), and LiOH (100 mg, 2.38 mmol) were added. The mixture was reacted at room temperature for 1 hour. The reaction solution was evaporated to dryness, and 0.3 mL of dilute hydrochloric acid was added. A large amount of yellow solid precipitated. The solid was filtered, evaporated to dryness, and 250 mg of yellow solid was obtained. That is, 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (250 mg, 0.46 mmol, 85.5%). m / z 548.18 [M+H] + .

[0181] Step 7: Preparation of {[(3S)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzimidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0182] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{1-methyl-7-[(3S)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (200 mg, 0.37 mmol), dimethylamine hydrochloride (60 mg, 0.74 mmol), methylimidazolium (70 mg, 0.85 mmol), and solvent acetonitrile (15 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (130 mg, 0.46 mmol) and react at room temperature for 0.5 h. Rotate the reaction mixture to dryness, add 10 mL of water to the reaction flask, and extract three times with 30 mL of dichloromethane / methanol mixture each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography (dichloromethane / methanol = 95 / 5) to obtain 150 mg of a yellow solid, namely {[(3S)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (150 mg, 0.26 mmol, 71.5%). m / z 575.18 [M+H] + .

[0183] Step 8: Preparation of 5-(5-amino-6-{7-[(3S)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (26)

[0184] In a 25 mL round-bottom flask, the reactant {[(3S)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (150 mg, 0.26 mmol) and solvent methanol (4 mL) were added, followed by the addition of 2 mL of 6N hydrochloric acid (12.00 mmol). The reaction mixture was reacted at room temperature for 1 hour. The reaction solution was evaporated to dryness, and 10 mL of sodium bicarbonate aqueous solution was added. The mixture was then extracted three times with a dichloromethane / methanol mixture, 30 mL each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 80 / 20) to obtain 65 mg of a yellow-brown solid. The product is 5-(5-amino-6-{7-[(3S)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (65 mg, 0.13 mmol, 51.0%). m / z 475.20 [M+H] + HPLC: 97.14%.

[0185] 1 H NMR (400MHz, DMSO) δ8.65(s,1H),8.18(s,2H),7.48(d,J=7.7Hz,1H),7.21(t,J=7.9Hz,1H),7.00(d,J=8.3Hz,2H),4.56(s,3H),3.91(s,3H), 3.24(d,J=11.3Hz,2H),3.16(s,1H),3.07(d,J=21.0Hz,7H),2.69-2.56(m,1H),2.39(t,J=10.6Hz,1H),2.00(d,J=12.3Hz,1H),1.81(s,2H).

[0186] Example 27: Preparation of 5-(5-amino-6-{7-[(3R)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (27)

[0187] Step 1: Preparation of {[(3R)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester}

[0188] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (4.4 g, 20.00 mmol), {[(3R)-hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.0 g, 20.00 mmol), cesium carbonate (13.0 g, 39.90 mmol), XPhos Pd G4 (1.7 g, 2.00 mmol), and solvent 1,4-dioxane (100 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (dichloromethane / methanol = 92 / 8) to give 5.5 g of a brown oily substance. This yielded {[(3R)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (5.5 g, 13.26 mmol, 66.2%). m / z 340.08 [M+H] + .

[0189] Step 2: Preparation of {[(3R)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0190] In a 250 mL round-bottom flask, the reactants {[(3R)-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.5 g, 13.26 mmol), methylamine hydrochloride (1.5 g, 22.22 mmol), potassium carbonate (7.7 g, 55.72 mmol), and solvent acetonitrile (100 mL) were added and stirred overnight at 70°C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 30 / 70) to obtain 4.0 g of crude brown oily product. That is, {[(3R)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (5.5 g, 16.21 mmol, 80.9%). m / z 351.1 [M+H] + .

[0191] Step 3: Preparation of {[(3R)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0192] In a 500 mL round-bottom flask, the reactant {[(3R)-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (4.0 g, 11.42 mmol), methanol (100 mL), and water (30 mL) were added. Sodium hydrosulfite (20 g, 114.88 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 2.4 g of a yellow oily substance, namely {[(3R)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.6 g, 8.11 mmol, 63.4%). m / z 321.2 [M+H] + .

[0193] Step 4: Preparation of {[(3R)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0194] In a 100 mL round-bottom flask, the reactants {[(3R)-1-[3-amino-2-(methylamino)phenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.6 g, 8.11 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (1.9 g, 8.72 mmol), methylimidazolium (1.5 g, 18.27 mmol), and solvent acetonitrile (100 mL) were added. TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (2.7 g, 9.62 mmol) was added, and the reaction was carried out at room temperature for 12 hours. Then, acetic acid (2 mL) was added to the reaction mixture, and the temperature was raised to 100 °C and stirred for 2 hours. The reaction mixture was rotated until acetonitrile was removed. The crude product was added to a saturated sodium bicarbonate aqueous solution (4 mL). The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined and evaporated to dryness. The crude product was purified by column chromatography (dichloromethane / methanol = 97 / 3) to give 1.5 g of a yellow solid, namely {[(3R)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.6 g, 5.18 mmol, 63.8%). m / z 502.0 / 504.0 [M+H] + .

[0195] Step 5: Preparation of methyl 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate

[0196] In a 100 mL round-bottom flask, add the reactants {[(3R)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (500 mg, 1.00 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)pyrazole-3-carboxylate (300 mg, 1.13 mmol), cesium carbonate (600 mg, 1.84 mmol), PdCl2 (dppf) (100 mg, 0.14 mmol), water (4 mL), and solvent 1,4-dioxane (20 mL). Stir the reaction mixture in N2 at 100 °C for about 3 hours. The reaction solution was evaporated to dryness, and the pH was adjusted with 3N dilute hydrochloric acid. Methanol and water (4 mL) were added, and the mixture was sonicated for 15 min. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to obtain 450 mg of a yellow solid, namely methyl 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate (450 mg, 0.80 mmol, 80.5%). m / z 562.14 [M+H] + .

[0197] Step 6: Preparation of 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0198] In a 25 mL round-bottom flask, methyl 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylate (600 mg, 1.07 mmol) and solvents MeOH (15 mL), water (3 mL), and LiOH (300 mg, 7.14 mmol) were added. The mixture was reacted at 70°C for 1 hour. The reaction solution was evaporated to dryness, and 0.3 mL of dilute hydrochloric acid was added. A large amount of yellow solid precipitated. The solid was filtered, evaporated to dryness, and 500 mg of yellow solid was obtained. That is, 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (500 mg, 0.91 mmol, 85.5%). m / z 548.18 [M+H] + .

[0199] Step 7: Preparation of {[(3R)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzimidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0200] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{1-methyl-7-[(3R)-3-({[(2-methylprop-2-yl)oxy]carbonyl}amino)hexahydropyridin-1-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (250 mg, 0.46 mmol), dimethylamine hydrochloride (50 mg, 0.61 mmol), methylimidazolium (100 mg, 1.22 mmol), and acetonitrile (15 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (160 mg, 0.57 mmol) and react at room temperature for 1 hour. Rotate the reaction mixture until no acetonitrile remains, add 10 mL of water to the reaction flask, and extract three times with 30 mL of dichloromethane / methanol mixture each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography (dichloromethane / methanol = 95 / 5) to obtain 200 mg of a yellow solid, namely {[(3R)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (200 mg, 0.35 mmol, 76.2%). m / z 575.18 [M+H] + .

[0201] Step 8: Preparation of 5-(5-amino-6-{7-[(3R)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (27)

[0202] In a 25 mL round-bottom flask, the reactant {[(3R)-1-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (200 mg, 0.35 mmol) and solvent methanol (4 mL) were added, followed by the addition of hydrochloric acid (2 mL, 24.00 mmol). The reaction mixture was reacted at room temperature for 1 hour. The reaction solution was evaporated to dryness, and 10 mL of sodium bicarbonate aqueous solution was added. The mixture was then extracted three times with a dichloromethane / methanol mixture, 30 mL each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 80 / 20) to obtain 92 mg of a yellow-brown solid. The product was 5-(5-amino-6-{7-[(3R)-3-aminohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (92 mg, 0.18 mmol, 52.9%). m / z 475.20 [M+H] + HPLC: 94.92%.

[0203] 1 H NMR (400MHz, DMSO) δ8.65(s,1H),8.18(s,2H),7.48(d,J=7.8Hz,1H),7.21(t,J=7.9Hz,1H),7.01(d,J=7.3Hz,2H),4.57(s,3H),3.91(s,3H) ,3.24(d,J=10.9Hz,2H),3.17(s,1H),3.07(d,J=21.7Hz,7H),2.70-2.53(m,1H),2.41(t,J=10.5Hz,1H),2.02(d,J=9.6Hz,1H),1.81(s,2H).

[0204] Example 28: Preparation of 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (28)

[0205] Step 1: Preparation of {[(3S,5S)-5-fluoro-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0206] In a 250 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (2.0 g, 9.09 mmol), {[(3S,5S)-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.0 g, 9.09 mmol), cesium carbonate (5 g, 15.35 mmol), XPhos Pd G4 (0.8 g, 0.91 mmol), and solvent 1,4-dioxane (100 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to give 2.4 g of a brown oil. This yielded {[(3S,5S)-5-fluoro-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.4 g, 6.72 mmol, 75.0%). m / z 358.18 [M+H] + .

[0207] Step 2: Preparation of {[(3S,5S)-5-fluoro-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0208] In a 100 mL round-bottom flask, the reactants {[(3S,5S)-5-fluoro-1-(2-fluoro-3-nitrophenyl)hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.4 g, 6.72 mmol), methylamine, hydrochloride (0.8 g, 11.85 mmol), potassium carbonate (3.2 g, 23.15 mmol), and solvent acetonitrile (70 mL) were added and stirred overnight at 65°C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (50 mL each time). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 2.2 g of crude brown oily product. That is, {[(3S,5S)-5-fluoro-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.2 g, 5.97 mmol, 88.0%). m / z 369.22 [M+H] + .

[0209] Step 3: Preparation of {[(3S,5S)-1-[3-amino-2-(methylamino)phenyl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0210] In a 250 mL round-bottom flask, the reactant {[(3S,5S)-5-fluoro-1-[2-(methylamino)-3-nitrophenyl]hexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (2.2 g, 5.97 mmol), methanol (50 mL), and water (15 mL) were added. Sodium hydrosulfite (12 g, 68.93 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 100 / 0) to obtain 1.3 g of a yellow oil. That is, {[(3S,5S)-1-[3-amino-2-(methylamino)phenyl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (1.3 g, 3.84 mmol, 65.0%). m / z 339.22 [M+H] + .

[0211] Step 4: Preparation of {[(3S,5S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0212] In a 100 mL round-bottom flask, add the reactants {[(3S,5S)-1-[3-amino-2-(methylamino)phenyl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (1.3 g, 3.84 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (1.0 g, 4.59 mmol), methylimidazolium (0.9 g, 10.96 mmol), and acetonitrile (40 mL) as solvent. Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (1.3 g, 4.63 mmol) and react at room temperature for 12 hours. Add acetic acid (2 mL) to the reaction mixture, heat to 100 °C, and stir for 2 hours. The reaction solution was rotated until acetonitrile was removed, dissolved in a small amount of ethanol, and sonicated for 15 min to obtain a suspension. The suspension was filtered, the filter cake was collected, and dried under reduced pressure to obtain 1.6 g of a gray solid, namely {[(3S,5S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (1.6 g, 3.07 mmol, 80.0%). m / z 520.11 / 522.17 [M+H] + .

[0213] Step 5: Preparation of 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0214] In a 100 mL round-bottom flask, add the reactants {[(3R,5R)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (1040 mg, 2.00 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)pyrazole-3-carboxylate (560 mg, 2.10 mmol), potassium carbonate (600 mg, 4.34 mmol), PdCl2 (dppf) (130 mg, 0.18 mmol), water (10 mL), and solvent 1,4-dioxane (40 mL). Stir the reaction mixture overnight in N2 at 100 °C. The reaction mixture was evaporated to dryness, and the pH was adjusted with 4N dilute hydrochloric acid. Methanol and water (10 mL) were added, and the mixture was sonicated for 15 min. The mixture was extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 85 / 15) to obtain 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (450 mg, 0.97 mmol, 48.3%). m / z 466.23 [M+H] + .

[0215] Step 6: Preparation of 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (28)

[0216] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (150 mg, 0.32 mmol), dimethylamine hydrochloride (50 mg, 0.61 mmol), methylimidazolium (70 mg, 0.85 mmol), and solvent CH3CN (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (120 mg, 0.43 mmol) and react at room temperature for 1 h. The reaction solution was rotated until acetonitrile-free. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 85 / 15) to obtain 80 mg of a yellow solid. The solid was dissolved in a small amount of methanol, sonicated for 15 min, filtered, and the filter cake was collected and dried under reduced pressure to obtain 64 mg of a yellow solid, namely 5-(5-amino-6-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (64 mg, 0.12 mmol, 36.9%). m / z 493.48 [M+H] + HPLC: 91.62%.

[0217] 1 H NMR (400MHz, DMSO) δ8.67(s,1H),8.22(s,2H),7.58(d,J=8.0Hz,1H),7.26(t,J= 7.9Hz,1H),7.10(d,J=7.7Hz,1H),7.03(s,1H),5.18(d,J=46.7Hz,1H),4.59(s, 3H),3.92(s,3H),3.81-3.69(m,1H),3.64(d,J=9.6Hz,1H),3.51(t,J=12.0Hz,1 H),3.20-2.97(m,7H),2.83-2.71(m,1H),2.49-2.41(m,1H),1.96-1.71(m,1H).

[0218] Example 29: Preparation of 3-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzi[d]imidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (29)

[0219] Step 1: Preparation of {[(3S,5S)-1-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzimidazol-4-yl)-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester

[0220] In a 100 mL round-bottom flask, add the reactants {[(3S,5S)-1-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (260 mg, 0.50 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)pyrazole-3-carboxylate (560 mg, 2.10 mmol), K2CO3 (170 mg, 1.23 mmol), PdCl2 (dppf) (70 mg, 0.10 mmol), water (5 mL), and solvent 1,4-dioxane (20 mL). Stir the reaction mixture overnight in N2 at 100 °C. The reaction solution was evaporated to dryness, and the residue was dissolved in water. The mixture was then extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to give 170 mg of a yellow solid, namely {[(3S,5S)-1-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzo[d]imidazol-4-yl)-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (170 mg, 0.29 mmol, 57.5%). m / z 592.33 [M+H] + .

[0221] Step 2: Preparation of 3-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzi[d]imidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (29)

[0222] In a 25 mL round-bottom flask, the reactant {[(3S,5S)-1-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzimidazol-4-yl)-5-fluorohexahydropyridin-3-yl]amino}methane-2-methylpropyl-2-yl ester (170 mg, 0.29 mmol) and solvent methanol (3 mL) were added, followed by hydrochloric acid (3 mL, 18.00 mmol). The reaction mixture was incubated at room temperature for 1 hour. The reaction solution was evaporated to dryness, and the reaction was monitored by LCMS to confirm completion. The reaction solution was evaporated to dryness. The crude product was dissolved in a small amount of methanol (3 mL), and the pH was adjusted with sodium bicarbonate aqueous solution. The solution was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 85 / 15) to give 17 mg of a yellow solid, namely the product 3-{7-[(3S,5S)-5-amino-3-fluorohexahydropyridin-1-yl]-1-methylbenzo[d]imidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (17 mg, 0.03 mmol, 11.7%). m / z 492.24 [M+H] + HPLC: 96.93%.

[0223] 1 H NMR(400MHz,DMSO)δ8.51(s,1H),8.37(s,1H),8.05(s,1H),7.94(s,2H),7.54(d ,J=8.0Hz,1H),7.24(t,J=7.9Hz,1H),7.07(d,J=7.6Hz,1H),5.13(d,J=46.6Hz,1 H),4.58(s,3H),4.56-4.42(m,1H),4.06-3.94(m,2H),3.63-3.42(m,6H),3.00(d d,J=39.6,12.8Hz,1H),2.43-2.24(m,1H),2.10-1.94(m,4H),1.72-1.44(m,1H).

[0224] Example 32: Preparation of 5-{5-amino-6-[7-(1-amino-3-azabicyclo[3.2.1]oct-3-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (32)

[0225] Step 1: Preparation of {[3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-1-yl]amino}methane-2-methylpropyl-2-yl ester}

[0226] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (1.0 g, 4.55 mmol), (3-azabicyclo[3.2.1]oct-1-ylamino)methane-2-methylpropyl-2-yl ester (1.0 g, 4.55 mmol), cesium carbonate (2.5 g, 7.67 mmol), XPhos Pd G4 (0.4 g, 0.46 mmol), and solvent 1,4-dioxane (50 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (ethyl acetate / petroleum ether = 48 / 52) to give 0.9 g of a brown oil. This yielded {[3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-1-yl]amino}methane-2-methylpropyl-2-yl ester (0.9 g, 2.46 mmol, 52.9%). m / z 366.22 [M+H] + .

[0227] Step 2: Preparation of 2-methylpropyl-2-yl ({3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane acid ester)

[0228] In a 100 mL round-bottom flask, the reactants {[3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-1-yl]amino}methane-2-methylpropyl-2-yl ester (900.0 mg, 2.46 mmol), methylamine hydrochloride (200 mg, 2.96 mmol), potassium carbonate (1000 mg, 7.24 mmol), and solvent acetonitrile (30 mL) were added and stirred overnight at 70°C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 0.8 g of crude brown oily product. That is, ({3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (0.8 g, 2.12 mmol, 88.9%). m / z 377.22 [M+H] + .

[0229] Step 3: Preparation of 2-methylpropyl-2-yl ({3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane acid-2-methylpropyl-2-yl ester

[0230] In a 100 mL round-bottom flask, the reactant ({3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (800 mg, 2.12 mmol), methanol (45 mL), and water (15 mL) were added. Sodium hydrosulfite (3000 mg, 17.23 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 80 / 20) to obtain 0.4 g of a yellow oily substance. That is, ({3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (0.4 g, 1.15 mmol, 57.1%). m / z 347.24 [M+H] + .

[0231] Step 4: Preparation of 2-methylpropyl-2-yl methanesulfonate ({{3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methanesulfonate

[0232] In a 100 mL round-bottom flask, add the reactants ({3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (400 mg, 1.15 mmol), 3-amino-6-bromopyrazine-2-carboxylic acid (300 mg, 1.38 mmol), methylimidazolium (300 mg, 3.65 mmol), and acetonitrile (20 mL) as solvent. Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (350 mg, 1.25 mmol) and react at room temperature for 2 hours. Add acetic acid (2 mL) to the reaction mixture, heat to 100 °C, and stir for 1 hour. The reaction solution was rotated until acetonitrile was removed, dissolved in methanol, filtered, and the filtrate was evaporated to dryness. The crude product was then added to a saturated sodium bicarbonate aqueous solution (15 mL), precipitating a large amount of yellow solid. The precipitate was filtered, and the filter cake was collected and dried in an oven to obtain 320 mg of the yellow solid, namely ({3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (320 mg, 0.61 mmol, 53.3%). m / z 528.11 / 530.22 [M+H] +

[0233] Step 5: Preparation of 5-(5-amino-6-{1-methyl-7-[1-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0234] In a 100 mL round-bottom flask, add the reactants ({3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (300 mg, 0.57 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)pyrazole-3-carboxylate (200 mg, 0.75 mmol), potassium carbonate (200 mg, 1.45 mmol), PdCl2 (dppf) (50 mg, 0.07 mmol), water (6 mL), and solvent 1,4-dioxane (30 mL). Stir the reaction mixture overnight in N2 at 100 °C. The reaction solution was evaporated to dryness, and the residue was dissolved in water. The mixture was then extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to give 300 mg of a yellow solid, namely 5-(5-amino-6-{1-methyl-7-[1-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (300 mg, 0.52 mmol, 92.1%). m / z 574.29 [M+H] + .

[0235] Step 6: Preparation of 2-methylpropyl-2-yl methane ester ({3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane ester)

[0236] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{1-methyl-7-[1-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (300 mg, 0.52 mmol), dimethylamine hydrochloride (80 mg, 0.98 mmol), methylimidazolium (100 mg, 1.22 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (180 mg, 0.64 mmol) and react at room temperature for about 3 h. The reaction solution was rotated until acetonitrile-free, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 95 / 15) to give 300 mg of a yellow solid, namely ({3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (300 mg, 0.50 mmol, 95.5%). m / z 601.33 [M+H] + .

[0237] Step 7: Preparation of 5-{5-amino-6-[7-(1-amino-3-azabicyclo[3.2.1]oct-3-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (32)

[0238] In a 25 mL round-bottom flask, the reactant ({3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-1-yl}amino)methane-2-methylpropyl-2-yl ester (300 mg, 0.50 mmol) and solvent methanol (4 mL) were added, followed by hydrochloric acid (2 mL, 64.66 mmol). The reaction was carried out at room temperature for 1 hour. The reaction solution was evaporated to dryness, and the reaction was monitored by LCMS, indicating that the reaction was complete. The reaction solution was evaporated to dryness. The crude product was dissolved in a small amount of methanol (3 mL), and the pH was adjusted with sodium bicarbonate aqueous solution. The solution was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 85 / 15) to give 200 mg of a yellow solid, namely the product 5-{5-amino-6-[7-(1-amino-3-azabicyclo[3.2.1]oct-3-yl)-1-methylbenzo[d]imidazol-2-yl]pyrazin-2-yl}-2,N,N-trimethylpyrazol-3-carboxamide (200 mg, 0.39 mmol, 78.3%). m / z 501.45 [M+H] +HPLC: 97.88%.

[0239] 1 H NMR (400MHz, DMSO) δ8.65 (s, 1H), 8.14 (s, 2H), 7.53 (dd, J = 5.6, 3.4Hz, 1H), 7.25-7 .22(m,2H),7.00(s,1H),4.65(s,3H),3.91(s,3H),3.16(s,1H),3.06(d,J=20.3Hz, 7H),2.88(d,J=10.2Hz,1H),2.79(d,J=10.0Hz,1H),2.35(s,1H),2.06-1.94(m,1H ),1.92-1.82(m,2H),1.78-1.68(m,1H),1.59-1.49(m,1H),1.45(d,J=10.5Hz,1H).

[0240] Example 33: Preparation of 5-(5-amino-6-{7-[(8-trans)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (33)

[0241] Step 1: Preparation of 2-methylpropyl-2-yl 8-trans-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane ester

[0242] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (1 g, 4.55 mmol), (8-trans)-3-azabicyclo[3.2.1]oct-8-ylcarbamate tert-butyl ester (0.86 g, 3.80 mmol), cesium carbonate (2.0 g, 6.14 mmol), XPhos Pd G4 (0.33 g, 0.38 mmol), and solvent XPhos Pd G4 (0.33 g, 0.38 mmol) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (ethyl acetate / petroleum ether = 48 / 52) to give 1.0 g of a brown oil. The crude product, 8-trans-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester, was obtained (1.0 g, 2.74 mmol, 71.4%). m / z 366.15 [M+H] + .

[0243] Step 2: Preparation of {[(8-trans)-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester}

[0244] In a 100 mL round-bottom flask, the reactant 8-trans-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (1.0 g, 2.74 mmol), methylamine, hydrochloride (0.4 g, 5.93 mmol), potassium carbonate (1.6 g, 11.58 mmol), and solvent acetonitrile (30 mL) were added and stirred overnight at 70 °C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with 50 mL of ethyl acetate each time. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 0.7 g of crude brown oily product. That is, {[(8-trans)-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (0.7 g, 1.86 mmol, 70.0%). m / z 377.22 [M+H] + .

[0245] Step 3: Preparation of {[(8-trans)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester}

[0246] In a 100 mL round-bottom flask, the reactant {[(8-trans)-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (0.7 g, 1.86 mmol), methanol (40 mL), and water (8 mL) were added. Sodium hydrosulfite (3 g, 17.23 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 80 / 20) to obtain 300 mg of a yellow oil. That is, {[(8-trans)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.87 mmol, 46.6%). m / z 347.24 [M+H] + .

[0247] Step 4: Preparation of {[(8-trans)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0248] In a 100 mL round-bottom flask, add the reactants 3-amino-6-bromopyrazine-2-carboxylic acid (218 mg, 1.00 mmol), {[(8-trans)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.87 mmol), methylimidazolium (200 mg, 2.44 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (300 mg, 1.07 mmol) and react at room temperature for 2 hours. Add acetic acid (2 mL) to the reaction mixture, heat to 100 °C, and stir for 1 hour. The reaction solution was rotated until acetonitrile was removed, dissolved in methanol, filtered, and the filtrate was evaporated to dryness. The crude product was then added to a saturated sodium bicarbonate aqueous solution (15 mL), precipitating a large amount of yellow solid. The precipitate was filtered, and the filter cake was collected and dried in an oven to obtain 300 mg of the yellow solid, namely {[(8-trans)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzi[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.57 mmol, 60.0%). m / z 528.11 / 530.22 [M+H] + .

[0249] Step 5: Preparation of 5-(5-amino-6-{1-methyl-7-[(8-trans)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0250] In a 100 mL round-bottom flask, add the reactants {[(8-trans)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.57 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)pyrazole-3-carboxylate (200 mg, 0.75 mmol), potassium carbonate (170 mg, 1.23 mmol), PdCl2 (dppf) (50 mg, 0.07 mmol), water (4 mL), and solvent 1,4-dioxane (16 mL). Stir the reaction mixture in N2 at 100 °C for about 2 h. The reaction solution was evaporated to dryness, and the residue was dissolved in water. The mixture was then extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to give 280 mg of a yellow solid, namely 5-(5-amino-6-{1-methyl-7-[(8-trans)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (280 mg, 0.49 mmol, 93.3%). m / z 574.29 [M+H] + .

[0251] Step 6: Preparation of {[(8-trans)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0252] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{1-methyl-7-[(8-trans)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (200 mg, 0.35 mmol), dimethylamine hydrochloride (60 mg, 0.74 mmol), methylimidazolium (90 mg, 1.10 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (130 mg, 0.46 mmol) and react overnight at room temperature. The reaction solution was rotated until acetonitrile-free, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 92 / 8) to give 150 mg of a yellow solid, namely {[(8-trans)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (150 mg, 0.25 mmol, 71.6%). m / z 601.33 [M+H] + .

[0253] Step 7: Preparation of 5-(5-amino-6-{7-[(8-trans)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (33)

[0254] Add the reactant {[(8-trans)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (150 mg, 0.25 mmol) and solvent methanol (5 mL) to a 25 mL round-bottom flask, add hydrochloric acid (2 mL), and react at room temperature for 3 hours. The reaction solution was evaporated to dryness. The crude product was dissolved in a small amount of methanol (3 mL), and the pH was adjusted with sodium bicarbonate aqueous solution. The solution was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 85 / 15) to give 50 mg of a yellow solid, namely the product 5-(5-amino-6-{7-[(8-trans)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (110 mg, 0.21 mmol, 84.0%). m / z 501.45 [M+H] +HPLC: 94.36%.

[0255] 1 H NMR (400MHz, DMSO) δ8.64(s,1H),8.11(s,2H),7.51(d,J=7.0Hz,1H),7.28-7.18(m,2H),7.00(s,1H),4.65(s,3 H),3.91(s,3H),3.14-3.01(m,9H),2.97(d,J=10.5Hz,2H),2.05(s,2H),2.01-1.91(m,2H),1.88-1.77(m,2H).

[0256] Example 34: Preparation of 5-(5-amino-6-{7-[(8-cis)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (34)

[0257] Step 1: Preparation of 2-methylpropyl-2-yl 8-cis-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane ester

[0258] In a 150 mL round-bottom flask, 1-bromo-2-fluoro-3-nitrobenzene (2 g, 9.09 mmol), 8-cis-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (2 g, 8.84 mmol), cesium carbonate (4.5 g, 13.81 mmol), XPhos Pd G4 (0.8 g, 0.93 mmol), and 1,4-dioxane solvent (80 mL) were added. The reaction mixture was stirred overnight at 110 °C under N2. The reaction mixture was filtered, the filtrate was evaporated to dryness, and purified by column chromatography (ethyl acetate / petroleum ether = 48 / 52) to give 1.3 g of a brown oil. The crude product, 8-cis-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester, was obtained (1.3 g, 3.56 mmol, 39.4%). m / z 366.15 [M+H] + .

[0259] Step 2: Preparation of {[(8-cis)-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0260] In a 100 mL round-bottom flask, the reactant 8-cis-3-(2-fluoro-3-nitrophenyl)-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (1.3 g, 3.56 mmol), methylamine, hydrochloride (0.4 g, 5.93 mmol), potassium carbonate (1.5 g, 10.85 mmol), and solvent acetonitrile (30 mL) were added and stirred overnight at 70 °C. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (50 mL each time). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The filtrate was purified by column chromatography (ethyl acetate / petroleum ether = 50 / 50) to obtain 0.8 g of crude brown oily product. That is, 8-cis-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (800 mg, 2.12 mmol, 61.5%). m / z 377.22 [M+H] + .

[0261] Step 3: Preparation of {(8-cis)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0262] In a 100 mL round-bottom flask, the reactant 8-cis-3-[2-(methylamino)-3-nitrophenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (0.8 g, 2.12 mmol), methanol (40 mL), and water (8 mL) were added. Sodium hydrosulfite (4 g, 22.98 mmol) was added in three portions at room temperature, and the mixture was stirred for approximately 0.5 h at room temperature. The reaction solution was filtered, and the filter cake was washed three times with a small amount of methanol. The filtrates and washings were combined, evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 80 / 20) to obtain 300 mg of a yellow oil. That is, {[(8-cis)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.87 mmol, 40.7%). m / z 347.24 [M+H] + .

[0263] Step 4: Preparation of {[(8-cis)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0264] In a 100 mL round-bottom flask, add the reactants 3-amino-6-bromopyrazine-2-carboxylic acid (200 mg, 0.92 mmol), {[(8-cis)-3-[3-amino-2-(methylamino)phenyl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (300 mg, 0.87 mmol), methylimidazolium (200 mg, 2.44 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (300 mg, 1.07 mmol) and react at room temperature for 2 hours. Add acetic acid (2 mL) to the reaction mixture, heat to 100 °C, and stir for 1 hour. The reaction solution was rotated until acetonitrile was removed, dissolved in methanol, filtered, and the filtrate was evaporated to dryness. The crude product was then added to a saturated sodium bicarbonate aqueous solution (15 mL), precipitating a large amount of yellow solid. The precipitate was filtered, and the filter cake was collected and dried in an oven to obtain 100 mg of yellow solid, namely {[(8-cis)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (100 mg, 0.19 mmol, 20.0%). m / z 528.11 / 530.22 [M+H] + .

[0265] Step 5: Preparation of 5-(5-amino-6-{1-methyl-7-[(8-cis)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid

[0266] In a 100 mL round-bottom flask, add the reactants {[(8-cis)-3-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (100 mg, 0.19 mmol), methyl 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)pyrazole-3-carboxylate (60 mg, 0.23 mmol), potassium carbonate (60 mg, 0.43 mmol), PdCl2 (dppf) (60 mg, 0.08 mmol), water (4 mL), and solvent 1,4-dioxane (16 mL). Stir the reaction mixture in N2 at 100 °C for about 2 h. The reaction solution was evaporated to dryness, and the residue was dissolved in water. The mixture was then extracted three times with dichloromethane / methanol (9 / 1, 60 mL). The organic phases were combined, evaporated to dryness, and the crude product was purified by column chromatography (dichloromethane / methanol = 93 / 7) to obtain 90 mg of a yellow solid, namely 5-(5-amino-6-{1-methyl-7-[(8-cis)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (90 mg, 0.16 mmol, 90.0%). m / z 574.29 [M+H] + .

[0267] Step 6: Preparation of {[(8-cis)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester

[0268] In a 100 mL round-bottom flask, add the reactants 5-(5-amino-6-{1-methyl-7-[(8-cis)-8-({[(2-methylprop-2-yl)oxy]carbonyl}amino)-3-azabicyclo[3.2.1]oct-3-yl]benzo[d]imidazol-2-yl}pyrazin-2-yl)-2-methylpyrazol-3-carboxylic acid (90 mg, 0.16 mmol), dimethylamine hydrochloride (30 mg, 0.37 mmol), methylimidazolium (40 mg, 0.49 mmol), and solvent acetonitrile (20 mL). Add TCFH N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (60 mg, 0.21 mmol) and react overnight at room temperature. The reaction solution was rotated until acetonitrile-free, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 92 / 8) to give 80 mg of a yellow solid, namely {[(8-cis)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (80 mg, 0.13 mmol, 84.9%). m / z 601.33 [M+H] + .

[0269] Step 7: Preparation of 5-(5-amino-6-{7-[(8-cis)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (34)

[0270] Add the reactant {[(8-cis)-3-[2-(3-amino-6-{5-[(dimethylamino)carbonyl]-1-methylpyrazol-3-yl}pyrazin-2-yl)-3-methylbenzo[d]imidazol-4-yl]-3-azabicyclo[3.2.1]oct-8-yl]amino}methane-2-methylpropyl-2-yl ester (80 mg, 0.13 mmol) and solvent methanol (1 mL) to a 25 mL round-bottom flask, then add 2 mL of hydrochloric acid methanol solution. React at room temperature for 3 hours. Since the reaction was not complete, continue reacting overnight. Processing will be done after returning to work on Monday. The reaction solution was evaporated to dryness. The crude product was dissolved in a small amount of methanol (3 mL), and the pH was adjusted with sodium bicarbonate aqueous solution. The solution was extracted three times with dichloromethane. The organic phases were combined, evaporated to dryness, and the crude product was purified by silica gel column chromatography (dichloromethane / methanol = 85 / 15) to give 20 mg of a yellow solid, namely the product 5-(5-amino-6-{7-[(8-cis)-8-amino-3-azabicyclo[3.2.1]oct-3-yl]-1-methylbenzo[d]imidazol-2-yl}pyrazin-2-yl)-2,N,N-trimethylpyrazol-3-carboxamide (20 mg, 0.04 mmol, 30.0%). m / z 501.45 [M+H]+ HPLC: 94.36%.

[0271] 1 H NMR (400MHz, DMSO) δ8.65(s,1H),8.12(s,2H),7.52(d,J=7.8Hz,1H),7.44(d,J=7.6Hz,1H),7.24(t,J=7.9Hz,1H),7.00(s,1H),4.66 (s,3H),3.92(s,3H),3.48(d,J=10.5Hz,2H),3.14-3.00(m,7H),2.77(dd,J=10.8,3.3Hz,2H),2.07-1.88(m,4H),1.84-1.72(m,2H).

[0272] Example 43: Preparation of 3-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]benzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (43)

[0273] Step 1: Preparation of 2-methylpropyl-2-yl(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine-1-carboxylic acid ester

[0274] tert-2-methyl(2S)piperazine-1-carboxylic acid tert-butyl ester (881.3 mg, 4.40 mmol), 3-bromo-2-fluoro-1-nitrobenzene (880.0 mg, 4 mmol), cesium carbonate (2606.4 mg, 8.00 mmol), Xant phos (462.9 mg, 0.80 mmol), and Pd2(dba)3 (366.3 mg, 0.40 mmol) were added to 1,4-dioxane (10 mL), substituted with N2 three times, and heated in an oil bath to 110 °C with stirring overnight. The reaction was quenched with water, extracted with ethyl acetate (100 mL x 3), the organic phases were combined and washed with saturated NaHCO3 (aq) and saturated brine; dried over anhydrous Na2SO4, filtered, concentrated, and the crude product was column-passed (EA:PE = 1:5) to give the target product 2-methylpropyl-2-yl(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine-1-carboxylic acid ester (650 mg, 1.92 mmol, 47.9%).

[0275] Step 2: Preparation of 2-methyl-2-ylpropyl(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl]piperazine-1-carboxylic acid ester

[0276] 2-Methylpropyl-2-yl(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine-1-carboxylate (650 mg, 1.92 mmol), methylamine hydrochloride (193.9 mg, 2.87 mmol), and K₂CO₃ (794.0 mg, 5.75 mmol) were added to 10 mL of DMF and reacted at 130 °C for 16 h. The reaction was monitored by TLC until complete. The reaction solution was added dropwise to water, resulting in the precipitation of a yellow solid. The mixture was filtered, the filter cake was washed with water, and dried to obtain 2-methyl-2-ylpropyl(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl]piperazine-1-carboxylate (490 mg, 1.40 mmol, 73.0%) as an orange-yellow solid. m / z 351.19 [M+H] + .

[0277] Step 3: Preparation of 2-methyl-2-ylpropyl(2S)-4-(2,3-diaminophenyl)-2-methylpiperazine-1-carboxylic acid ester

[0278] In a 100 mL single-necked flask, 2-methyl-2-ylpropyl(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl]piperazine-1-carboxylate (1.7 g, 4.85 mmol), Pd / C (0.17 g, 1.60 mmol), and solvent ethanol (20 mL) were added. Hydrazine hydrate (5 mL, 51.40 mmol) was added to the reaction mixture, and the mixture was stirred at 80 °C for 1 hour. The reaction solution changed from yellow to colorless. The reaction was monitored for completeness by TLC. The mixture was filtered through a diatomaceous earth liner, and the filtrate was diluted with water. The aqueous layer was extracted with EA. The EA layers were combined and washed with saturated NaCl solution, dried over Na₂SO₄, filtered, and concentrated. The solution was then passed through a column with EA:PE 1:1 elution. The sample 2-methyl-2-ylpropyl(2S)-4-(2,3-diaminophenyl)-2-methylpiperazine-1-carboxylate (500 mg, 1.63 mmol, 33.6%) was collected.

[0279] Step 4: Preparation of 2-methylpropyl-2-yl(2S)-4-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]-2-methylpiperazine-1-carboxylic acid ester

[0280] 2-Methyl-2-ylpropyl(2S)-4-(2,3-diaminophenyl)-2-methylpiperazine-1-carboxylic acid ester (617.3 mg, 1.93 mmol), HOAt (524.4 mg, 3.85 mmol), and EDCI (738.7 mg, 3.85 mmol) were added to 10 mL of DMF and reacted overnight at room temperature. The reaction was monitored by LCMS until complete, quenched with water, filtered, and the filter cake was dried. The crude product was added to acetic acid (5 mL) and reacted at 130 °C for 1 hour. The reaction was quenched with water, extracted with EA, and the reaction solution was concentrated. The crude product was passed through a column and eluted with EA:PE = 1:4 to obtain 2-methylpropyl-2-yl(2S)-4-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]-2-methylpiperazine-1-carboxylic acid ester (340 mg, 0.68 mmol, 21.1%) as a yellow solid.

[0281] Step 5: Preparation of 2-methylpropyl-2-yl(2S)-4-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzimidazol-4-yl)-2-methylpiperazine-1-carboxylic acid ester

[0282] 2-Methylpropyl-2-yl(2S)-4-[2-(3-amino-6-bromopyrazin-2-yl)-3-methylbenzimidazol-4-yl]-2-methylpiperazine-1-carboxylic acid ester (340 mg, 0.68 mmol), 1,1′-bis(di-cyclohexylphosphino)ferrocene palladium dichloride (51.2 mg, 0.07 mmol), and K2CO3 (280.6 mg, 2.03 mmol) were added to 1,4-dioxane (10 mL) and H2O (2 mL). The mixture was stirred overnight at 85 °C under N2 protection. The reaction solution was directly concentrated, mixed with a silica gel plate, and eluted with EA:PE = 1:1 to obtain 2-methylpropyl-2-yl(2S)-4-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzimidazol-4-yl)-2-methylpiperazine-1-carboxylic acid ester (250 mg, 0.44 mmol, 64.4%) as a yellow solid.

[0283] Step 6: Preparation of 3-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]benzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (43)

[0284] In a 50 mL single-necked flask, add the reactants 2-methylpropyl-2-yl(2S)-4-(2-{3-amino-6-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-yl}-3-methylbenzimidazol-4-yl)-2-methylpiperazine-1-carboxylic acid ester (250 mg, 0.44 mmol), hydrochloric acid (1 mL, 32.33 mmol), and solvent MeOH (2 mL). The reaction solution was stirred at room temperature for 2 hours, then concentrated. The pH was adjusted to alkaline with 1N NaOH, resulting in the precipitation of a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain 3-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]benzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (220 mg, 0.45 mmol, 103.9%). m / z 474.2 [M+H] + .

[0285] 1 H NMR(400MHz)δ8.49(s,1H),8.36(s,1H),8.05(s,1H),7.95(s,2H),7.46(d,J=7.9 Hz,1H),7.19(t,J=7.8Hz,1H),7.01(d,J=7.6Hz,1H),4.58(s,3H),4.54-4.40(m,1 H),3.99(d,J=11.3Hz,2H),3.55-3.45(m,2H),3.14(d,J=10.3Hz,2H),2.99(s,3H) ,2.68(s,1H),2.43-2.29(m,1H),2.21(s,1H),2.02(s,4H),1.04(d,J=5.7Hz,3H).

[0286] Example 44: Preparation of 3-{7-[(3S)-3,4-dimethylpiperazin-1-yl]-1-methylbenzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (44)

[0287] 3-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]benzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (50 mg, 0.11 mmol), iodomethane (0.01 mL, 0.11 mmol), and K₂CO₃ (43.8 mg, 0.32 mmol) were added to DMF (2 mL), and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by LC-MS until complete, and the reaction was quenched with water. The aqueous layer was extracted with EA (3 × 50 mL). The EA layers were combined and washed with water (3 × 50 mL). The mixture was dried over anhydrous Na₂SO₄, filtered, and concentrated. The crude product was subjected to thin-layer chromatography and developed with EA:PE 1:1 to give 3-{7-[(3S)-3,4-dimethylpiperazin-1-yl]-1-methylbenzimidazol-2-yl}-5-[1-(3,4,5,6-tetrahydro-2H-pyran-4-yl)pyrazol-4-yl]pyrazin-2-amine (40 mg, 0.08 mmol, 76.4%) as a yellow solid. m / z 488.22M+H] + .

[0288] 1 H NMR (400MHz, DMSO) δ8.50(s,1H),8.37(s,1H),8.05(s,1H),7.93(s,2H),7.48(d,J=7.9H z,1H),7.20(t,J=7.9Hz,1H),7.04(d,J=7.7Hz,1H),4.58(s,3H),4.51-4.44(m,1H),3.99 (d,J=11.3Hz,2H),3.50(s,2H),3.17(s,2H),3.04(d,J=7.3Hz,1H),2.94(d,J=11.6Hz,2H ),2.56(s,1H),2.47-2.40(m,1H),2.34(s,3H),2.05-1.99(m,4H),1.10(d,J=6.0Hz,3H).

[0289] Biological evaluation

[0290] Example A: Test of the inhibitory activity of the compounds of the present invention against MNK1 and MNK2 kinases.

[0291] Using ADP-Glo TMThe kinase assay kit (purchased from Promega, catalog number V9101) was used to test the inhibitory effects of the compounds on MNK1 and MNK2, respectively. Both kinase reactions were performed in reaction buffer (15 mM HEPES pH 7.4, 20 mM NaCl, 1 mM EGTA, 10 mM MgCl2, 0.1 mg / mL BGG, and 0.02% Tween-20). The BGG (Bovine Gamma Globulin) solution was purchased from Absin Biotech (Shanghai) Co., Ltd. (catalog number abs47047684), and HEPES, NaCl, EDTA, MgCl2, and Tween-20 were all purchased from Sangon Biotech (Shanghai) Co., Ltd. (hereinafter referred to as "Sangon"). Aside from the different MNK kinases used, both the MNK1 and MNK2 reaction systems contained different final concentrations of MNK substrate peptide (Ac-TATKSGSTTKNR-NH2, ordered from Sangon Biotech) and ATP (Order NO. A600560, purchased from Sangon Biotech). Specifically, the MNK1 reaction system ultimately contained 10 nM recombinant MNK1 kinase (purchased from Carna Biosciences, catalog number 02-145), 100 μM MNK substrate peptide, 300 μM ATP, and different final concentrations of the test compound; the MNK2 reaction system ultimately contained 10 nM recombinant MNK2 kinase (purchased from Carna Biosciences, catalog number 02-146), 200 μM MNK substrate peptide, 10 μM ATP, and different final concentrations of the test compound. The final concentration gradients of the test compounds were 1, 10, 100, 1000, and 10000 nM.

[0292] Before the test, MNK1 (or MNK2) kinase was prepared to 50 nM using reaction buffer. The test compound was diluted with DMSO to prepare gradient solutions of 100, 1000, 10000, 100000, and 1000000 nM, and then each DMSO solution was diluted 20-fold with reaction buffer. For the MNK1 reaction system, the MNK substrate peptide was prepared to 500 μM and ATP to 750 μM using reaction buffer; for the MNK2 reaction system, the MNK substrate peptide was prepared to 1000 μM and ATP to 25 μM using reaction buffer.

[0293] During testing, 1 μL each of the prepared MNK1 (or MNK2) kinase, compound, and MNK substrate peptide solution were added to a 384-well white flat-bottomed polystyrene plate. After incubation at room temperature for 10 minutes, 2 μL of the corresponding ATP solution was added. After further incubation at room temperature for 40 minutes, 5 μL of ADP-Glo ​​was added to each well. TMThe reagent was added and incubated for another 40 minutes, then the reaction was stopped. A luminescent signal for kinase activity readings was generated by adding 10 μL of kinase detection reagent and incubating for 40 minutes. The fluorescence signal at a wavelength of 560 nm was detected using a Tecan Spark microplate reader.

[0294] Using wells without compounds as negative controls (Y) and wells without enzymes as positive controls (Z), the inhibition rate of each compound against the corresponding kinase was calculated based on the fluorescence signal (X) of each test well, using the following formula: Inhibition (%) = 100% - (XZ) / (YZ) × 100%. The half-maximal inhibitory concentration (IC50) of each compound against the corresponding kinase was then calculated using GraphPad 7.0 software. 50 ).

[0295] Table 3: Activity detection results of compounds MNK1 and MNK2 of the present invention

[0296] Table 3 shows that the compounds of the present invention have IC50 values ​​of less than 5 nM for both kinases MNK1 and MNK2, and exhibit excellent inhibitory effects on MNK1 and MNK2 activity. They can be used to prepare drugs for diseases related to these effects.

[0297] Example B: Pharmacokinetic Test of the Compounds of the Invention in Mice

[0298] Eight-week-old female BALB / c mice (purchased from Zhuhai Beston Biotechnology Co., Ltd.) were used, with six animals per group for each compound. The compounds of this invention were formulated in a solvent containing 5% DMSO, 60% PEG400, and 35% purified water. The dosage was 10 mg / kg, administered via a single gavage. Blood was collected alternately from the fundus venous plexus at time points: before administration and 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after administration. Approximately 0.06 mL of blood was collected in centrifuge tubes (anticoagulated with EDTA), centrifuged at 3200 g for 10 minutes at 4°C, and the supernatant plasma was separated and stored at -20°C for analysis. Several plasma samples were extracted with acetonitrile, and the concentration of the compounds in the plasma was determined using liquid chromatography-mass spectrometry (LC-MS / MS). Pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on the average plasma drug concentration data at different time points. The data are summarized in Table 4.

[0299] Table 4: Pharmacokinetic characteristics of the compounds of the present invention in mice after oral administration

[0300] Among them, the control molecule 01 is a compound described in the literature "Discovery of a Selective and Potent Inhibitor of Mitogen-Activated Protein Kinase-Interacting Kinases 1 and 2 (MNK1 / 2) Utilizing Structure-Based Drug Design", and its structural formula is as follows:

[0301] As the results above show, the representative compounds of this invention exhibited good in vivo exposure after oral administration to mice, especially Examples 4, 6, 9, 18, 28, 29, 32, and 34, whose plasma exposure (AUC) and peak concentration (Cmax) were significantly higher than those of the control molecule O1. These pharmacokinetic characteristics indicate that the in vivo exposure level of these representative compounds after oral administration reached an unexpected level, potentially leading to superior antitumor activity.

[0302] Example C: Distribution test of the compound of the present invention in mouse tissue

[0303] Twenty-four 8-week-old female BALB / c mice (purchased from Zhuhai Beston Biotechnology Co., Ltd.) were used, with six animals per group for each compound. The compounds of this invention were prepared in a solvent containing 5% DMSO, 60% PEG400, and 35% purified water. The dosage was 10 mg / kg, administered by gavage once. Blood was collected 1 h and 4 h after administration, and heart, liver, lung, and kidney tissues were collected immediately after sacrifice. Approximately 0.1 mL of blood was collected in a centrifuge tube (anticoagulated with EDTA), centrifuged at 3200g for 10 minutes at 4°C, and the supernatant plasma was separated. A number of tissue samples were weighed, added to a certain amount of phosphate-buffered saline (PBS) solution, homogenized, and frozen at -20°C for later analysis. Several plasma or tissue samples were extracted with acetonitrile, and the concentration of the compounds in the samples was determined using liquid chromatography-mass spectrometry (LC-MS / MS). Data summaries are shown in Tables 5 and 6.

[0304] Table 5: Tissue distribution of the compound of the present invention in mice 1 hour after oral administration.

[0305] Table 6: Tissue distribution of the compounds of the present invention in mice 4 hours after oral administration

[0306] The control molecule 01 is as described in Example B.

[0307] As can be seen from the above results, after oral administration to mice, the representative compounds of the present invention are mainly distributed in solid tissues such as the liver, lungs and kidneys. The ratio of drug concentration in tissues to plasma is significantly greater than 1, indicating that the drug has a stronger affinity for tissues. Moreover, its tissue distribution is significantly higher than that of the control molecule O1, indicating that compared with the control molecule O1, the compounds of the present invention have a stronger affinity for tissues and have the potential to exert a stronger therapeutic effect in these tissues.

[0308] Example D: Distribution test of the compound of the present invention in mouse tissue

[0309] Four female nude mice (purchased from Guangdong Zhiyuan Biomedical Technology Co., Ltd.), approximately 6 weeks old, were subcutaneously inoculated with NCI-H446 cells (2×10⁶ cells) after one week of acclimatization. 6 One per site, until the subcutaneous tumor reaches approximately 500 mm². 3 Subsequently, the compounds of this invention were prepared in a solvent containing 5% DMSO, 60% PEG400, and 35% purified water. The dosage was 10 mg / kg. Blood and tumor tissue were collected 2 hours after gavage administration. Approximately 0.1 mL of blood was collected in a centrifuge tube (anticoagulated with EDTA), centrifuged at 3200g for 10 minutes at 4°C, and the supernatant plasma was separated. A certain amount of tissue was weighed, added to a certain amount of phosphate-buffered saline (PBS) solution, homogenized, and frozen at -20°C for later analysis. A certain amount of plasma or tissue samples were transferred, extracted with acetonitrile, and the concentration of the compounds in the samples was determined using liquid chromatography-mass spectrometry (LC-MS / MS). The data are summarized in Table 7.

[0310] Table 7: Tumor tissue distribution 2 hours after oral administration of the compounds of the present invention

[0311] As can be seen from the above results, the representative compound described in this invention exhibits tumor-targeting properties after oral administration to mice, with a drug concentration ratio of 5.2 in tumor tissue to plasma, indicating that the drug is highly enriched at the tumor site, providing a strong material basis for enhancing anti-tumor efficacy.

[0312] Furthermore, in preliminary efficacy tests, the representative compound of this invention significantly reduced tumor volume after oral administration to mice, further demonstrating that the compound of this invention can target tumors and exert a significant effect, a result that was unexpected.

[0313] Industrial applicability

[0314] The compounds of the present invention have excellent MNK1 and / or MNK2 inhibitory effects and can be used as drugs for treating, preventing or improving diseases related to these effects.

[0315] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound of formula (I), or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotopic derivative thereof: in, R1is selected from a 5-membered heteroaryl; said heteroaryl has 1-3 heteroatoms selected from N, O, S; said heteroaryl is optionally substituted with one or more R 1A R 1A is selected from C 1-6 alkyl, -C(O)NR 1B R 1C , 4-8 membered heterocycloalkyl; said heterocycloalkyl contains 1 heteroatom O; R 1B and R 1C are each independently selected from hydrogen, C 1-6 alkyl, halogen, cyano, amino, carboxyl, hydroxyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy; R3is independently selected from hydrogen, 4-12 membered heterocycloalkyl, 4-12 membered heterocyclyl; said heterocycloalkyl, heterocyclyl contains 1-4 heteroatoms selected from N, O, S; any carbon atom of said heterocycloalkyl, heterocyclyl can be oxo; said heterocycloalkyl, heterocyclyl is unsubstituted or substituted with one or more R 3A each R 3A is independently selected from hydrogen, C 1-6 alkyl, halogen, cyano, amino, carboxyl, hydroxyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy; R5is independently selected from hydrogen, -C(O)NR 5B R 5C ; R 5B and R 5C each independently selected from hydrogen, C 3-10 cycloalkyl.

2. The compound according to claim 1, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, characterized in that: R1is selected from 5-membered heteroaryl; said heteroaryl has 2 heteroatoms selected from N, O, S; said heteroaryl can be optionally substituted with one or more R 1A substituents; Preferably, R1is selected from optionally substituted 1A substituted pyrrole, pyrazole, imidazole, furan, thiophene, isoxazole, triazole, isothiazole, thiazole, oxazole ring; More preferably, R1is selected from optionally substituted pyrazole ring, imidazole ring, thiazole ring; 1A substituted pyrazole ring, imidazole ring, thiazole ring; Also preferably, R1is selected from optionally substituted 1A substituted Also preferably, R1is selected from optionally substituted 1A substituted Each R 1A Each is independently selected from C 1-3 Alkyl, C 4-6 Alkyl, -C(O)NR 1B R 1C 4-6 membered heterocyclic alkyl groups, 7-8 membered heterocyclic alkyl groups; wherein the heterocyclic alkyl group contains one heteroatom O; Preferably, each R 1A each independently selected from the group consisting of methyl, ethyl, propyl, isopropyl, -C(O)NR 1B R 1C , 4-5 membered heterocycloalkyl, 6-7 membered heterocycloalkyl; the heterocycloalkyl contains 1 heteroatom O; It is also preferred that each R 1A each independently is selected from the group consisting of methyl, -C(O)NR 1B R 1C a 6-membered heterocycloalkyl; said heterocycloalkyl contains 1 heteroatom O; R 1B , R 1C are each independently selected from the group consisting of hydrogen, C 1-3 alkyl, C 4-6 alkyl, fluorine, chlorine, bromine, iodine, cyano, amino, carboxyl, hydroxyl, nitro, C 1-3 alkoxy, C 4-6 alkoxy, C 1-3 haloalkyl, C 4-6 haloalkyl, C 1-3 haloalkoxy, C 4-6 haloalkoxy; Preferably, R 1B R 1C Each is independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, C4 alkyl, C5 alkyl, C6 alkyl, fluorine, chlorine, cyano, amino, carboxyl, hydroxyl, methoxy, ethoxy, propoxy, isopropoxy, C4 alkoxy, C5 alkoxy, C6 alkoxy, halomethyl, haloethyl, halopropyl, haloisopropyl, C4 haloalkyl, C5 haloalkyl, C6 haloalkyl, halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, C4 haloalkoxy, C5 haloalkoxy, C6 haloalkoxy; Preferably, R 1B , R 1C are each independently selected from hydrogen, methyl; It is also preferred that each R 1A each independently is selected from the group consisting of methyl, It is also preferred that each R 1A each independently is selected from the group consisting of methyl, Most preferably, R1 is selected from Most preferably, R1 is selected from 3. The compound according to claim 1, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, characterized in that: R3is selected from hydrogen, 3-4 membered heterocycloalkyl, 5-12 membered heterocycloalkyl, 4-5 membered heterocyclyl, 6-12 membered heterocyclyl; said heterocycloalkyl, heterocyclyl contains 1-3 heteroatoms selected from N, O, S; any carbon atom of said heterocycloalkyl, heterocyclyl can be oxidized; said heterocycloalkyl, heterocyclyl is unsubstituted or substituted with one or more R 3A substituents; Preferably, R3is selected from hydrogen, 5-6 membered heterocycloalkyl, 7-8 membered heterocycloalkyl, 9-10 membered heterocycloalkyl, 5-6 membered heterocyclyl, 7-8 membered heterocyclyl, 9-10 membered heterocyclyl; said heterocycloalkyl, heterocyclyl contains 1-3 heteroatoms selected from N, O, S; any carbon atom of said heterocycloalkyl, heterocyclyl can be oxo; said heterocycloalkyl, heterocyclyl is unsubstituted or substituted by one or more R 3A substituents; Preferably, R3 is selected from hydrogen, 5-membered heterocyclic alkyl, 6-membered heterocyclic alkyl, 7-membered heterocyclic alkyl, 8-membered heterocyclic alkyl, and 6-membered heterocyclic group; Preferably, R3 is selected from hydrogen, unsubstituted hydrogen, or hydrogen ions containing one or more R atoms. 3A Replacement Preferably, R3 is selected from hydrogen, unsubstituted hydrogen, or hydrogen ions containing one or more R atoms. 3A Replacement each R is independently selected from hydrogen, C 3A each R is independently selected from hydrogen, C 1-3 alkyl, C 4-6 alkyl, F, CI, Br, I, CN, NH2, COOH, OH, NO2, C 1-3 alkoxy, C 4-6 alkoxy, C 1-3 haloalkyl, C 4-6 haloalkyl, C 1-3 haloalkoxy, C 4-6 haloalkoxy; Preferably, each R 3A Each is independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, C4 alkyl, C5 alkyl, C6 alkyl, fluorine, chlorine, cyano, amino, carboxyl, hydroxyl, methoxy, ethoxy, propoxy, isopropoxy, C4 alkoxy, C5 alkoxy, C6 alkoxy, halomethyl, haloethyl, halopropyl, haloisopropyl, C4 haloalkyl, C5 haloalkyl, C6 haloalkyl, halomethoxy, haloethoxy, halopropoxy, haloisopropoxy, C4 haloalkoxy, C5 haloalkoxy, C6 haloalkoxy; More preferably, each R 3A each independently is selected from hydrogen, methyl, fluorine, chlorine, amino; Most preferably, R3 is selected from hydrogen, Most preferably, R3 is selected from hydrogen, 4. The compound according to claim 1, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, characterized in that: R5is selected from hydrogen, -C(O)NR 5B R 5C ; R 5B and R 5C are each independently selected from hydrogen, C 3-6 cycloalkyl, C 7- 8cycloalkyl, C 9-10 cycloalkyl; R is preferably selected from hydrogen, C 5B R is preferably selected from hydrogen, C 5C each independently selected from hydrogen, C 3-4 cycloalkyl, C 5-6 cycloalkyl; More preferably, R 5B and R 5C each independently is selected from the group consisting of hydrogen, cyclopropane, cyclobutane, cyclopentane, cyclohexane; Most preferably, R5 is selected from hydrogen, 5. The compound according to any one of claims 1-4, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, characterized in that: The compound of formula (I) has the structure shown in formula (II): The definitions of R1 and R3 are as described in any one of claims 1, 2 or 3; Alternatively, the compound of formula (I) has the structure shown in formula (III): wherein R1, R 5B or R 5C are as defined in any one of claims 1, 2 or 4; Alternatively, the compound of formula (I) has the structure shown in formula (Ⅳ-1), (Ⅳ-2), or (Ⅳ-3): R3 is defined as described in claim 1 or 3.

6. The compound according to any one of claims 1-5, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, or isotope derivative, characterized in that: The compound is selected from the following compounds:

7. A pharmaceutical composition comprising the compound of any one of claims 1-6, or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, and pharmaceutically acceptable auxiliary ingredient.

8. Use of any compound, prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, or pharmaceutical composition of any one of claims 1-6 in the preparation of a medicament for treating and / or preventing and / or improving diseases related to the biological activity of MNK1 and / or MNK2; Preferably, the diseases associated with the biological activity of MNK1 and / or MNK2 are selected from cancer and hyperproliferative diseases.

9. A method of treating and / or preventing and / or improving a disease associated with the biological activity of MNK1 and / or MNK2, comprising administering to a patient in need a therapeutically effective amount of any one of claims 1-6, or a prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, or pharmaceutical composition of claim 7. Preferably, the diseases associated with the biological activity of MNK1 and / or MNK2 are selected from cancer and hyperproliferative diseases.

10. The compound, or its prodrug, solvate, crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, metabolite, isotope derivative, or pharmaceutical composition of any one of claims 1-6, for the treatment and / or prevention and / or improvement of diseases related to the biological activity of MNK1 and / or MNK2; Preferably, the diseases associated with the biological activity of MNK1 and / or MNK2 are selected from cancer and hyperproliferative diseases.

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