WO2021018287A1

WO2021018287A1 - Spiroaromatic compound, preparation and application thereof

Info

Publication number: WO2021018287A1
Application number: PCT/CN2020/106214
Authority: WO
Inventors: 郑乾刚; 曾庆龙; 许明
Original assignee: 上海奕拓医药科技有限责任公司
Priority date: 2019-08-01
Filing date: 2020-07-31
Publication date: 2021-02-04
Also published as: CN114269746A; CN112300160A

Abstract

Provided is a compound with inhibitory activity on SHP2, a preparation method therefor, and a use thereof. Specifically, provided is a compound represented by formula (I), a pharmaceutically acceptable salt thereof, a solvate, an isotopically substituted compound, a polymorph, a prodrug, or a metabolite thereof. The definition of each group is as described in the specification. The compound has high inhibitory activity on SHP2 and therefore can be used to prevent or treat SHP2-related diseases.

Description

A spiroaromatic compound, its preparation and application

Technical field

The invention discloses a spiroaromatic ring compound, its pharmaceutically acceptable salt, or its solvate, isotope substitution, prodrug or metabolite. The present invention also provides a method for preparing the compound, a pharmaceutical composition containing the compound, and the use of the compound for preparing a medicine for treating diseases or disorders related to abnormal SHP2 activity.

Background technique

Protein tyrosine phosphatase SHP2 occupies an extremely important position in the process of cell signal transduction, and is a target for the development of treatment of major diseases such as diabetes, autoimmune diseases and cancer. SHP2 is mutated or highly expressed in many diseases, such as Noonan Syndrome, Leopard Syndrome, Juvenile Myelomonocytic Leukemia, Neuroblastoma, Melanoma, Acute Myeloid Leukemia, Breast cancer, esophageal cancer, lung cancer, colon cancer, head cancer, neuroblastoma, squamous cell carcinoma of the head and neck, gastric cancer, anaplastic large cell lymphoma and glioblastoma, etc. Molecular biology studies have shown that SHP2 is involved in multiple tumor cell signal transduction pathways, such as MAPK, JAK/STAT and PI3K/Akt. At the same time, SHP2 is also responsible for the signal transduction of the PD1-PDL1 immunosuppressive pathway. Therefore, inhibiting the activity of SHP2 can reverse the immunosuppression in the tumor microenvironment.

SHP2 is composed of two N-terminal Src homology 2 domains (N-SH2 and C-SH2) and a protein tyrosine phosphatase catalytic domain (PTP). In the self-inhibition state, N-SH2 binds to PTP to form a ring structure, thereby hindering the binding of PTP to the substrate, so that the enzyme catalytic activity is inhibited; when the tyrosine of the upstream receptor protein is phosphorylated, N- SH2 binds to it, and the PTP catalytic domain is released to exert phosphatase activity.

At present, the development of SHP2 inhibitors is mainly based on allosteric inhibitors in the non-catalytic region, such as some compounds disclosed in WO2015107493A1, WO2016203404A1, WO2016203406A1, WO2017216706A1, WO2017211303A1, CN201710062495, WO2018136265A1WO2018057884, etc. Recent studies have shown that SHP2, as a novel druggable target, has attracted more and more attention. Therefore, there is an urgent need in this field to develop SHP2 inhibitors with novel structure, good biological activity and high druggability.

Summary of the invention

The object of the present invention is to provide a compound of formula I or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof, and the compound or the pharmaceutical composition in the prevention and treatment of SHP2 Application in abnormally related diseases or conditions.

The first aspect of the present invention provides a compound represented by formula I, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof:

among them,

R _{1 is} selected from H, -halogen, -CN, -OH, -NO ₂ , HSO ₃ -, unsubstituted or halogenated C1-C6 alkylsulfonyl, unsubstituted or halogenated C1-C6 alkylcarboxy, Unsubstituted or halogenated C1-C6 alkylamino, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C1-6 alkoxy Carbonyl, unsubstituted or halogenated C1-6 alkylcarbonyl, unsubstituted or halogenated C2-C6 alkenylcarbonyl, unsubstituted or halogenated C1-C6 alkoxy-O-C1-C6 alkyl, unsubstituted Substituted or substituted 3-8 membered cycloalkyl, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl, unsubstituted or substituted 5-10 membered heteroaryl; The heterocyclic group or heteroaryl group contains 1-4 heteroatoms selected from the group consisting of N, O or S;

R _{2 is} selected from H, -halogen, amino, unsubstituted or substituted C1-C6 alkylamino, unsubstituted or substituted C1-C6 alkyl;

R _{3 is} selected from H, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted 5-10 membered aryl, substituted or unsubstituted 5 -10 membered heteroaryl; said heterocyclic group or heteroaryl group contains 1-4 heteroatoms selected from the group consisting of N, O or S;

X and Y are each independently selected from N or CR ₄ ;

R _{4 is} selected from H, halogen, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, (substituted or unsubstituted C1-C6 Alkyl)SO-, (substituted or unsubstituted C1-C6 alkyl)SO ₂ -;

Or R ₄ and adjacent

Together to form a substituted or unsubstituted 5-10 membered aromatic ring, a substituted or unsubstituted 5-10 membered heteroaromatic ring or a substituted or unsubstituted 5-10 membered heterocyclic ring; the ring may contain 0-4 options Heteroatoms from the following group: N, O or S; the condition is that X and Y cannot be N at the same time;

Represents a single bond or a double bond;

Any of the above "substituted" means that one or more hydrogen atoms on the group are substituted by a substituent selected from the following group: -D, halogen, -OH, -NO ₂ , -NH ₂ , -N (unsubstituted or (Halogenated C1-C6 alkyl) ₂ , -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C1-C8 alkoxy Group-C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C3-C8 cycloalkyl-C1-C8 alkyl, unsubstituted or halogenated C1-C6 alkane Carbonyl, unsubstituted or halogenated C1-C6 alkoxycarbonyl, hydroxamic acid group, unsubstituted or halogenated C1-C6 alkyl mercapto, -S(O) ₂ N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -S(O) ₂ unsubstituted or halogenated C1-C6 alkyl, -N (unsubstituted or halogenated C1-C6 alkyl) S(O) ₂ N (unsubstituted Or halogenated C1-C6 alkyl) ₂ , -S(O)N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -S(O) (unsubstituted or halogenated C1-C6 alkyl) ), -N (unsubstituted or halogenated C1-C6 alkyl) S(O)N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -N (unsubstituted or halogenated C1-C6 alkane Group) S(O) (unsubstituted or halogenated C1-C6 alkyl), unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or A halogenated 4-8 membered saturated heterocyclic ring or carbocyclic ring; wherein the heteroaryl group contains 1-4 heteroatoms selected from the group consisting of N, O or S, and the heterocyclic ring contains 1-4 One heteroatom selected from the group: N, O or S.

As a preferred embodiment, the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof is shown in the following formula II:

among them,

X and Y are each independently selected from N or C;

R _4a and R _4b are each independently selected from none, H, halogen, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, ( Substituted or unsubstituted C1-C6 alkyl) SO-, (substituted or unsubstituted C1-C6 alkyl) SO ₂ -;

The definition of other substituents is as described above.

As another preferred embodiment, the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof is shown in the following formula III :

among them,

X and Y are independently selected from N or C; X and Y cannot be N at the same time;

Ring A is a substituted or unsubstituted 5-10 membered aromatic ring, a substituted or unsubstituted 5-10 membered heteroaromatic ring or a substituted or unsubstituted 5-10 membered heterocyclic group; the ring may contain 0-4 A heteroatom selected from the following group: N, O or S;

The above-mentioned "substituted" means that one or more hydrogen atoms on ring A are substituted by a substituent selected from the following group: -D, halogen, -OH, -NO ₂ , -NH ₂ , -NH (unsubstituted or halogenated C1-C6 alkyl), -N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -CN, halogenated or unsubstituted C1-C6 alkyl, halogenated or unsubstituted C1-C6 alkoxy Group, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO-, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -;

The definition of other substituents is as described above.

As a further preferred embodiment, the ring A is selected from: substituted or unsubstituted three-membered (hetero) ring, substituted or unsubstituted four-membered (hetero) ring, substituted or unsubstituted five-membered (hetero) ring , Substituted or unsubstituted six-membered (hetero) ring, substituted or unsubstituted five-membered aromatic (hetero) ring, substituted or unsubstituted six-membered aromatic (hetero) ring, substituted or unsubstituted seven-membered aromatic (hetero) Ring, substituted or unsubstituted five-membered six-membered (hetero) ring, substituted or unsubstituted six-membered six-membered (hetero) ring; the heterocyclic ring may contain 1-4 heteroatoms selected from the following group : N, O or S; The ring A can be optionally substituted, and the substituent is selected from: halogen, -CN, -OH, -NH ₂ , (halogenated or unsubstituted C1-C6 alkyl) ₂ N-, halogenated or unsubstituted C1-C6 alkyl, halogenated or unsubstituted C1-C6 alkoxy, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO- , (Halogenated or unsubstituted C1-C6 alkyl) SO ₂ -.

As a further preferred manner, the ring A is selected from any one of the following groups:

The above-mentioned ring A can be optionally substituted, and the substituents are selected from halogen, -CN, -OH, -NH ₂ , (halogenated or unsubstituted C1-C6 alkyl) ₂ N-, halogenated or unsubstituted C1-C6 alkyl, halogenated or unsubstituted C1-C6 alkoxy, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO-, (halogenated or unsubstituted C1 -C6 alkyl)SO ₂ -.

As a further preferred mode, in the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotope substitutions, polymorphs, prodrugs or metabolites thereof, R _{1 is} selected from H, halogenated or unsubstituted C1-C6 alkylcarbonyl, halogenated or unsubstituted C2-C6 alkenylcarbonyl, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -, substituted or Unsubstituted benzenesulfonyl, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl, unsubstituted or substituted 5-10 membered heteroaryl group; where the benzenesulfonyl group, carbocyclic group, heterocyclic group, aryl group and heteroaryl group are substituted, the number of substituents is 1-4, selected from H, NH ₂ , Halogen, halogenated or unsubstituted C3-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl and (halogenated or unsubstituted C1-C6 alkyl) ₂ N-.

As a further preferred mode, the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, R _{1 is} selected from H, halogenated or unsubstituted C1-C6 alkylcarbonyl, halogenated or unsubstituted C2-C6 alkenylcarbonyl, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -, substituted Or an unsubstituted benzenesulfonyl group, and any of the following groups that may be optionally substituted:

Wherein W is selected from C, O, S or N; wherein, when the benzenesulfonyl group and the optionally substituted ring group are substituted, the number of substituents is 1-4, selected from H, NH ₂ , Halogen, halogenated or unsubstituted C1-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl, and (halogenated or unsubstituted C1-C6 alkyl) ₂ N-.

As a further preferred mode, in the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotope substitutions, polymorphs, prodrugs or metabolites thereof, R _{3 is} selected from H, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl group and unsubstituted or substituted 5- 10-membered heteroaryl group; when the carbocyclic group, heterocyclic group, aryl group and heteroaryl group are substituted, the number of substituents is 1-4, selected from H, NH ₂ , halogen, halogenated Or unsubstituted C3-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) ₂ N- and halogenated or unsubstituted C1-C6 alkane基-NH-.

As a further preferred mode, the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, R _{3 is} selected from H and any of the following cyclic groups which may be optionally substituted:

Wherein W is selected from O, S or N; wherein, when the cyclic group is substituted, the number of substituents is 1-4, and the substituents are selected from H, NH ₂ , halogen, unsubstituted or halogenated C1- C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) ₂ N- and halogenated or unsubstituted C1-C6 alkyl -NH-.

As a further preferred mode, the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, R _{2 is} selected from H, -halogen, unsubstituted or substituted C1-C6 alkylamino, unsubstituted or substituted C1-C6 alkyl.

As a further preferred mode, the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, is selected from the following structures:

The second aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention for:

(a) Preparation of drugs for the prevention or treatment of diseases or disorders related to abnormal SHP2 activity;

(b) Preparation of drugs for preventing or treating SHP2-mediated diseases or conditions;

(c) Preparation of inhibitor drugs that inhibit SHP2 activity;

(d) Non-therapeutic inhibition of SHP2 activity in vitro;

(e) Non-therapeutic inhibition of tumor cell proliferation in vitro; and/or

(f) Treatment of diseases or conditions related to abnormal SHP2.

In a preferred embodiment, the disease related to abnormal SHP2 activity is selected from the following group: cancer; preferably Noonan syndrome, leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma , Acute myeloid leukemia, breast cancer, esophageal cancer, lung cancer, colon cancer, head cancer, neuroblastoma, squamous cell carcinoma of the head and neck, gastric cancer, anaplastic large cell lymphoma or glioblastoma.

The third aspect of the present invention provides a pharmaceutical composition, the pharmaceutical composition comprising:

(i) An effective amount of the compound of formula I, or a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, solvate, isotope substitution, prodrug or metabolite thereof ;with

(ii) A pharmaceutically acceptable carrier.

The fourth aspect of the present invention provides a method for inhibiting the activity of SHP2, said method comprising the steps of: administering an inhibitory effective amount of the compound of formula I as described in the first aspect of the present invention or its pharmaceutically acceptable The accepted salt, or an effective amount of the pharmaceutical composition according to the third aspect of the present invention is administered to the subject of inhibition.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, I will not repeat them here.

Detailed ways

After long-term and in-depth research, the inventors prepared a new type of allosteric inhibitor compound represented by formula I, which can "lock" the self-inhibition with weak SHP2 activity by binding to the non-catalytic region of SHP2 State, so as to achieve the purpose of inhibiting its activity. The compound of the present invention exhibits good biological activity and druggability, and has a good prospect for drug development. It has an inhibitory effect on SHP2 at a very low concentration (which can be as low as 100nM/L). The activity is quite excellent, so it can be used to treat SHP2 related diseases or disorders, such as tumors. Based on the above findings, the inventor completed the present invention.

the term

Unless otherwise defined, all scientific and technological terms herein have the same meanings as commonly understood by those skilled in the art to which the subject of the claims belongs. Unless otherwise specified, all patents, patent applications, and publications cited in this article are incorporated herein by reference in their entirety.

It should be understood that the above brief description and the following detailed description are exemplary and only used for explanation, without any limitation on the subject of the present invention. In this application, unless specifically stated otherwise, the use of the singular number also includes the plural number. It must be noted that unless clearly stated otherwise in the context, the singular form used in this specification and claims includes the plural form of the things referred to. It should also be noted that the use of "or" and "or" means "and/or" unless stated otherwise. In addition, the term "including" and other forms, such as "including", "including" and "containing" are not restrictive, and they can be open, semi-closed and closed. In other words, the term also includes the meaning of "essentially composed of" or "consisting of".

The definitions of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise specified, conventional methods within the technical scope of the art, such as mass spectrometry, NMR, IR, and UV/VIS spectroscopy and pharmacological methods, are used. Unless specific definitions are provided, the terms used in the descriptions of analytical chemistry, synthetic organic chemistry, and pharmaceuticals and medicinal chemistry herein are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and treatment of patients. For example, the manufacturer's instructions for the use of the kit can be used, or the reaction and purification can be performed in a manner known in the art or the instructions of the present invention. Generally, the above-mentioned techniques and methods can be implemented according to the descriptions in a number of summary and more specific documents cited and discussed in this specification according to conventional methods well known in the art. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural parts and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH ₂ O- is equivalent to -OCH ₂ -.

The chapter headings used in this article are only for the purpose of organizing the article, and should not be construed as a limitation on the subject. All documents or document parts cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and theses, are incorporated herein by reference in their entirety.

Certain chemical groups defined herein are preceded by simplified symbols to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the simplified notation does not include carbons that may be present in the substituents of the group.

In addition to the foregoing, when used in the specification and claims of this application, unless otherwise specified, the following terms have the following meanings.

In this application, the term "halogen" means fluorine, chlorine, bromine or iodine.

"Hydroxy" refers to the -OH group.

"Hydroxyalkyl" refers to an alkyl group as defined below that is substituted with a hydroxyl group (-OH).

"Carbonyl" refers to the -C(=O)- group.

"Nitro" refers to -NO ₂ .

"Cyano" refers to -CN.

"Amino" refers to -NH ₂ .

"Substituted amino" refers to an amino group substituted with one or two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl groups as defined below, for example, monoalkylamino, dialkylamino, alkyl Amido, aralkylamino, heteroaralkylamino.

"Carboxy" refers to -COOH.

In this application, as a group or a part of other groups (for example, used in halogen-substituted alkyl groups and other groups), the term "alkyl" refers to a fully saturated linear or branched hydrocarbon chain group, It consists only of carbon atoms and hydrogen atoms, has, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms, and is connected to the rest of the molecule through a single bond, such as including but not limited to Methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl , N-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl and decyl, etc. For the purposes of the present invention, the term "alkyl" refers to an alkyl group containing 1 to 8 carbon atoms.

In this application, as a group or a part of other groups, the term "alkenyl" means consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 20 (preferably 2 to 10 One, more preferably 2 to 6) carbon atoms and a straight or branched hydrocarbon chain group connected to the rest of the molecule through a single bond, such as but not limited to vinyl, propenyl, allyl, but- 1-alkenyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl, etc.

In this application, as a group or a part of other groups, the term "cycloalkyl" or "carbocyclyl" means a stable non-aromatic monocyclic or polycyclic hydrocarbon group composed of only carbon atoms and hydrogen atoms, It may include a fused ring system, a bridged ring system or a spiro ring system, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, and it is saturated or unsaturated It can be connected to the rest of the molecule through a single bond via any suitable carbon atom. Unless specifically indicated otherwise in this specification, the carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Exemplary cyclic hydrocarbon groups or carbocyclic groups include C3-C8 cycloalkyl groups. Furthermore, examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl , 1H-indenyl, 2,3-indanyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro- 7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzo ring Octenyl, fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2 .2]octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantane Group, octahydro-4,7-methylene-1H-indenyl and octahydro-2,5-methylene- and cyclopentadienyl.

In this application, as a group or part of another group, the term "heterocyclic group" means a group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur Stable 3- to 20-membered non-aromatic cyclic group. Unless otherwise specified in this specification, the heterocyclic group may be a monocyclic, bicyclic, tricyclic or more ring system, which may include a fused ring system, a bridged ring system or a spiro ring system; in the heterocyclic group The nitrogen, carbon, or sulfur atoms of may be optionally oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group can be connected to the rest of the molecule via a carbon atom or a heteroatom and through a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purpose of the present invention, the heterocyclic group is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur The group is more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]non Alkyl-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptan-2-yl, aza Cyclobutanyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxopentyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, Imidazolidinyl, quinazinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indoline, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl , Phthalimide, etc.

In this application, as a group or a part of other groups, the term "aryl" means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms (preferably having 6 to 10 carbon atoms). For the purpose of the present invention, the aryl group can be a monocyclic, bicyclic, tricyclic or more cyclic ring system, and can also be fused with a cycloalkyl or heterocyclic group as defined above, provided that the aryl group passes through The atoms on the aromatic ring are connected to the rest of the molecule through a single bond. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthryl, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazine-3(4H)-one-7-yl and the like.

In this application, the term "arylalkyl" refers to the above-defined alkyl group substituted by the above-defined aryl group.

In this application, as a group or part of another group, the term "heteroaryl" means having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 nitrogen atoms in the ring. A 5- to 16-membered conjugated ring system group of heteroatoms of, oxygen and sulfur. Unless otherwise specified in this specification, heteroaryl groups can be monocyclic, bicyclic, tricyclic or more cyclic ring systems, and can also be fused with cycloalkyl or heterocyclic groups as defined above, provided that the hetero The aryl group is connected to the rest of the molecule via a single bond through an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group can be optionally oxidized; the nitrogen atom can be optionally quaternized. For the purpose of the present invention, the heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably contains 1 to 4 selected heteroatoms. A stable 5- to 10-membered aromatic group from heteroatoms of nitrogen, oxygen, and sulfur or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, Benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indazinyl, isoindolyl, indazolyl, isoindazolyl , Purinyl, Quinolinyl, Isoquinolinyl, Diazonaphthyl, Naphthyridinyl, Quinoxolinyl, Pteridinyl, Carbazolyl, Carboline, Phenanthridinyl, Phenanthrolinyl, Acridine Group, phenazine group, isothiazolyl, benzothiazolyl, benzothienyl, oxtriazolyl, cinnolinyl, quinazolinyl, phenylthio, indolizinyl, phenanthryl, Isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thienyl, naphthopyridyl, [1,2,4]triazolo[4 ,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1, 2,4]Triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine Wait.

In this application, the term "heteroarylalkyl" refers to the above-defined alkyl group substituted by the above-defined heteroaryl group.

In this application, "optionally" or "optionally" means that the event or condition described later may or may not occur, and the description includes both occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl group" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups. The "optionally" substituents described in the claims and specification of the present invention are selected from alkyl, alkenyl, alkynyl, halogen, halogenated alkyl, halogenated alkenyl, halogenated alkynyl, cyano, nitro , Optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclic alkyl.

"SHP2" refers to "SrcHomolgy-2 phosphatase", also known as SH-PTP2, SH-PT3, Syp, PTP1D, PTP2C, SAP-2 or PTPN11.

The terms "part", "structural part", "chemical part", "group", and "chemical group" as used herein refer to specific fragments or functional groups in a molecule. The chemical moiety is generally considered to be a chemical entity embedded or attached to a molecule.

Conventional techniques for preparing/separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives using, for example, chiral high performance liquid chromatography) ), for example, see Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; AMStalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3 :341-63,2010; Fumis et al.(eds.),VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED.,Longman Scientific and Technical Ltd.,Essex,1991,809-816;Heller,Acc.Chem .Res.1990,23,128.

The present invention also includes all suitable isotopic variants of the compounds of the present invention or pharmaceutically acceptable salts thereof. Isotopic variants of the compound of the present invention or a pharmaceutically acceptable salt thereof are defined as those in which at least one atom is replaced by an atom having the same atomic number but having an atomic mass different from the atomic mass often found in nature. The isotopes that can be incorporated into the compounds of the present invention and their pharmaceutically acceptable salts include, but are not limited to, isotopes of H, C, N, and O, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ³⁵ S, ¹⁸ F, ³⁶ Cl and ¹²⁵ I. Isotopic variants of the compounds of the present invention or pharmaceutically acceptable salts thereof can be prepared by conventional techniques using appropriate isotopic variants of appropriate reagents.

In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic acid or an organic acid that can retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include, but are not limited to, formate, acetate, and 2,2-dichloroacetate , Trifluoroacetate, propionate, caproate, caprylate, caprate, undecylenate, glycolate, gluconate, lactate, sebacate, hexanoate Acid salt, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate , Cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate , Alginate, ascorbate, salicylate, 4-aminosalicylate, naphthalene disulfonate, etc. These salts can be prepared by methods known in the art.

"Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include but are not limited to the following salts: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins , Such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, bicyclic Hexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperazine Pyridine, N-ethylpiperidine, polyamine resin, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the art.

In the present application, "pharmaceutical composition" refers to a preparation of a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (such as a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of the organism, which is conducive to the absorption of the active ingredient and thus the biological activity.

The term "pharmaceutically acceptable" as used herein refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of the present invention, and is relatively non-toxic, that is, the substance can be administered to an individual without causing undesirable biological activity. Reacts or interacts in an undesirable manner with any components included in the composition.

In this application, "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvants, carriers, excipients, glidants, enhancers that are approved by the relevant government administration as acceptable for human or livestock use. Sweeteners, diluents, preservatives, dyes/colorants, flavors, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

The "tumor" of the present invention includes but is not limited to Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, sarcoma, melanoma, articular chondroma, cholangiomas, leukemia, breast cancer, stomach Intestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, esophageal cancer, pancreatic cancer, lung squamous cell carcinoma, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervix Cancer, ovarian cancer, bowel cancer, nasopharyngeal cancer, brain cancer, bone cancer, kidney cancer, oral cancer/head cancer, neuroblastoma, squamous cell carcinoma of the head and neck, anaplastic large cell lymphoma, or glioblast Tumors and other diseases.

The terms "preventive", "preventing" and "preventing" as used herein include reducing the likelihood of a disease or condition from occurring or worsening.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) Preventing the occurrence of a disease or condition in a mammal, especially when such a mammal is susceptible to the disease or condition, but has not been diagnosed as having the disease or condition;

(ii) Inhibiting the disease or disease, that is, curbing its development;

(iii) Alleviate the disease or condition, that is, make the state of the disease or condition subside; or

(iv) Alleviate the symptoms caused by the disease or condition.

The term "effective amount", "therapeutically effective amount" or "pharmaceutical effective amount" as used herein refers to at least one agent or compound that is sufficient to relieve one or more symptoms of the disease or condition being treated after administration The amount. The result can be a reduction and/or alleviation of signs, symptoms or causes, or any other desired changes in the biological system. For example, the "effective amount" for treatment is the amount of the composition containing the compound disclosed herein that is required to provide significant disease relief clinically. Techniques such as dose escalation tests can be used to determine the effective amount suitable for any individual case.

The terms "administration", "administration", "administration" and the like as used herein refer to methods capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, transduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Those skilled in the art are familiar with the application techniques that can be used for the compounds and methods described herein, for example in Goodman and Gilman, The Pharmaceutical Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical (current edition), Mack Publishing Co., Those discussed in Easton, Pa. In a preferred embodiment, the compounds and compositions discussed herein are administered orally.

The terms "drug combination", "drug combination", "combination", "administration of other treatments", "administration of other therapeutic agents" and the like as used herein refer to drug treatments obtained by mixing or combining more than one active ingredient. It includes fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, combination or sequential administration of at least one compound described herein and at least one synergistic agent to a patient in the form of separate entities. These also apply to cocktail therapy, such as the administration of three or more active ingredients.

Those skilled in the art should also understand that in the methods described below, the functional group of the intermediate compound may need to be protected by an appropriate protecting group. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl (e.g. tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl) , Tetrahydropyranyl, benzyl, etc. Suitable protecting groups for amino, amidino and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable protecting groups for mercapto include -C(O)-R" (wherein R" is an alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable carboxyl protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups can be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protective groups is detailed in Greene, T.W. and P.G.M. Wuts, Protective Groups in Organi Synthesis, (1999), 4th Ed., Wiley. The protecting group can also be a polymer resin.

Formula I compound

The present invention provides a compound represented by formula I, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof:

among them,

X and Y are each independently selected from N or CR ₄ ;

Or R ₄ and adjacent

Together to form a substituted or unsubstituted 5-10 membered aromatic ring, substituted or unsubstituted 5-10 membered heteroaromatic ring, substituted or unsubstituted 5-10 membered heterocyclic ring, or substituted or unsubstituted 5-10 membered ring Carbocyclic ring; the ring may contain 0-4 heteroatoms selected from the group consisting of N, O or S; the condition is that X and Y cannot be N at the same time;

Represents a single bond or a double bond;

Any of the above "substituted" means that one or more hydrogen atoms on the group are substituted by a substituent selected from the following group: -D, halogen, -OH, -NO ₂ , -NH ₂ , -N (unsubstituted or (Halogenated C1-C6 alkyl) ₂ , -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C1-C8 alkoxy, unsubstituted or halogenated C1-C8 alkoxy Group-C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C3-C8 cycloalkyl-C1-C8 alkyl, unsubstituted or halogenated C1-C6 alkane Carbonyl group, unsubstituted or halogenated C1-C6 alkoxycarbonyl, hydroxamic acid group, unsubstituted or halogenated C1-C6 alkyl mercapto group, -S(O) ₂ N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -S(O) ₂ unsubstituted or halogenated C1-C6 alkyl, -N (unsubstituted or halogenated C1-C6 alkyl) S(O) ₂ N (unsubstituted Or halogenated C1-C6 alkyl) ₂ , -S(O)N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -S(O) (unsubstituted or halogenated C1-C6 alkyl) ), -N (unsubstituted or halogenated C1-C6 alkyl) S(O)N (unsubstituted or halogenated C1-C6 alkyl) ₂ , -N (unsubstituted or halogenated C1-C6 alkane Group) S(O) (unsubstituted or halogenated C1-C6 alkyl), unsubstituted or halogenated 5-8 membered aryl, unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or A halogenated 4-8 membered saturated heterocyclic ring or carbocyclic ring; wherein the heteroaryl group contains 1-4 heteroatoms selected from the group consisting of N, O or S, and the heterocyclic ring contains 1-4 One heteroatom selected from the group: N, O or S.

Preferably, in formula I, when R ₄ is adjacent to

When jointly forming a substituted or unsubstituted 5-10 membered aromatic ring, substituted or unsubstituted 5-10 membered heteroaromatic ring, substituted or unsubstituted 5-10 membered heterocyclic ring, the ring formed is any one of the implementations herein The ring A described in the scheme.

The present invention provides a compound of Formula II, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof:

among them,

X and Y are each independently selected from N or C;

The definition of other substituents is as described above.

The present invention provides a compound of Formula III, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof:

among them,

Ring A is a substituted or unsubstituted 5-10 membered aromatic ring, a substituted or unsubstituted 5-10 membered heteroaromatic ring, a substituted or unsubstituted 5-10 membered heterocyclic ring, or a substituted or unsubstituted 5-10 membered ring Carbocyclic ring; said ring may contain 0-4 heteroatoms selected from the group consisting of N, O or S;

The definition of other substituents is as described above.

In the compound represented by formula III provided by the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, preferably, the ring A is selected from Self-substituted or unsubstituted three-membered (hetero) ring, substituted or unsubstituted four-membered (hetero) ring, substituted or unsubstituted five-membered (hetero) ring, substituted or unsubstituted six-membered (hetero) ring, substituted Or unsubstituted five-membered aromatic (hetero) ring, substituted or unsubstituted six-membered aromatic (hetero) ring, substituted or unsubstituted seven-membered aromatic (hetero) ring, substituted or unsubstituted five-membered and six-membered (hetero) ) Ring, substituted or unsubstituted six-membered and six-membered (hetero) ring; the heterocyclic ring may contain 1-4 heteroatoms selected from the group consisting of N, O or S; the ring A may be Optional substitution, the substituent is selected from: halogen, -CN, -OH, -NH ₂ , (halogenated or unsubstituted C1-C6 alkyl) ₂ N-, halogenated or unsubstituted C1-C6 alkyl , Halogenated or unsubstituted C1-C6 alkoxy, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO-, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -.

In the compound represented by formula III provided by the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, preferably, the ring A is Choose from any one of the following groups:

The compound represented by formula III provided by the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, more preferably, the ring A Any one selected from the following group:

More preferably, in the compound represented by formula III of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, ring A is optionally substituted :

More preferably

Wherein, when substituted, the substituent may be 1-2 substituents selected from halogen, C1-C6 alkyl and halogenated C1-C6 alkyl.

The compounds represented by formulas I, II and III provided by the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, preferably, the R _{1 is} selected from H, halogenated or unsubstituted C1-C6 alkylcarbonyl, halogenated or unsubstituted C2-C6 alkenylcarbonyl, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -, Substituted or unsubstituted benzenesulfonyl group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 5-10 membered aryl group, unsubstituted or Substituted 5-10 membered heteroaryl group; wherein the benzenesulfonyl group, carbocyclic group, heterocyclic group, aryl group and heteroaryl group are substituted, the number of substituents is 1-4, and the substituents are selected From H, NH ₂ , halogen, halogenated or unsubstituted C1-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) ₂ N-.

The compounds represented by formulas I, II and III provided by the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, preferably, the R _{1 is} selected from H, halogenated or unsubstituted C1-C6 alkylcarbonyl, halogenated or unsubstituted C2-C6 alkenylcarbonyl, (halogenated or unsubstituted C1-C6 alkyl) SO ₂ -, A substituted or unsubstituted benzenesulfonyl group, and any of the following groups which may be optionally substituted:

Preferably, R ₁ in formula I, II and III is selected from: unsubstituted C1-C6 alkyl-SO ₂ -, unsubstituted C2-C6 alkenyl carbonyl, unsubstituted or substituted 3-8 membered heterocycle Group, unsubstituted or substituted 3-8 membered cycloalkyl, unsubstituted or substituted 5-10 membered aryl, unsubstituted or substituted 5-10 membered heteroaryl; wherein, the heteroaryl, cycloalkane When the group, aryl group and heteroaryl group are substituted, the substituents are 1-3 substituents selected from halogen, unsubstituted C1-C6 alkyl and halogenated C1-C6 alkyl. More preferably, the heterocyclic group, cycloalkyl group, aryl group and heteroaryl group in R ₁ may be selected from pyrazinyl, phenyl, azetidinyl, cyclobutanyl, oxetanyl And pyridyl, these groups can be optionally substituted with 1 or 2 C1-C6 alkyl groups.

Among the compounds represented by formulas I, II and III provided by the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, preferably, R ₂ It is selected from H, halogen, amino and unsubstituted C1-C6 alkyl; more preferably, R _{2 is} selected from H and amino.

The compounds represented by formulas I, II and III provided by the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, preferably, the R _{3 is} selected from H, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5-10 membered heteroaryl; wherein the ring group can be optionally substituted, the number of the substituent is 1-4; the substituent is selected from H, NH ₂ , halogen, halogenated or Unsubstituted C1-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) ₂ N- and halogenated or unsubstituted C1-C6 alkyl -NH-.

Preferably, the compounds represented by formulas I, II and III provided by the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, R _{3 is} selected from H,

Wherein W is selected from O, S or N, and the cyclic group can be optionally substituted, the number of substituents is 1-4, and the substituents are selected from H, NH ₂ , halogen, unsubstituted or halogenated C1 -C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) ₂ N- and halogenated or unsubstituted C1-C6 alkyl -NH-.

More preferably, the compounds represented by formulas I, II and III of the present invention, or pharmaceutically acceptable salts thereof, or solvates, isotopic substitutions, polymorphs, prodrugs or metabolites thereof, R _{3 is} selected from 5-10 membered heteroaryl groups optionally substituted by amino and halogen, preferably 5-10 membered nitrogen-containing heteroaryl groups, more preferably pyridyl optionally substituted by amino and halogen, more preferably any Select substituted by amino and halogen:

In a particularly preferred embodiment, the compound represented by formula III of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, R _{1 is} selected From unsubstituted C1-C6 alkyl-SO ₂ -, unsubstituted C2-C6 alkenyl carbonyl, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 3-8 membered cycloalkyl, Unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5-10 membered heteroaryl group; wherein, when the heteroaryl group, cycloalkyl group, aryl group and heteroaryl group are substituted, the substituent is 1-3 substituents selected from halogen, unsubstituted C1-C6 alkyl and halogenated C1-C6 alkyl; more preferably, the heterocyclic group, cycloalkyl group, aryl group and heteroaryl group in R ₁ Can be selected from pyrazinyl, phenyl, azetidinyl, cyclobutanyl, oxetanyl and pyridinyl, and these groups can be optionally substituted by 1 or 2 selected from halogen and C1- Substituents of C6 alkyl; more preferably, R _{1 is} selected from optionally substituted with 1 or 2 substituents selected from halogen and C1-C6 alkyl:

R _{2 is} selected from H, halogen, amino and unsubstituted C1-C6 alkyl, more preferably selected from H and amino; R _{3 is} selected from: 5-10 membered heteroaryl optionally substituted by amino and halogen, preferably A 5- to 10-membered nitrogen-containing heteroaryl group is more preferably a pyridyl group optionally substituted with 1 or 2 substituents selected from amino and halogen, and more preferably the following substituted with 2 substituents selected from amino and halogen Group:

Ring A is optionally substituted:

In a particularly preferred embodiment, the compound represented by formula II of the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, R _{1 is} selected From unsubstituted C1-C6 alkyl-SO ₂ -, unsubstituted C2-C6 alkenyl carbonyl, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 3-8 membered cycloalkyl, Unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5-10 membered heteroaryl group; wherein, when the heteroaryl group, cycloalkyl group, aryl group and heteroaryl group are substituted, the substituent is 1-3 substituents selected from halogen, unsubstituted C1-C6 alkyl and halogenated C1-C6 alkyl; more preferably, the heterocyclic group, cycloalkyl group, aryl group and heteroaryl group in R ₁ Can be selected from pyrazinyl, phenyl, azetidinyl, cyclobutanyl, oxetanyl and pyridinyl, these groups can be optionally substituted with 1 or 2 C1-C6 alkyl groups ; R _{2 is} selected from H, halogen, amino and unsubstituted C1-C6 alkyl; R _{3 is} selected from: 5-10 membered heteroaryl optionally substituted by amino and halogen, preferably 5-10 membered nitrogen-containing hetero Aryl is more preferably pyridyl optionally substituted with amino and halogen; X is CH; Y is N.

Here, the wavy line on the group indicates the position where the group is attached to the rest of the compounds of formula I, II and III.

It should be understood that in the present invention, the selection of W, X and Y should satisfy the bond valence theory, and when the bond valence theory is not satisfied, a hydrogen atom is used to make up. For example, depending on the ring, W can be -CH-, -CH ₂ -or -NH-, and X or Y can be CH.

The compound provided by the present invention, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug, or metabolite thereof is selected from the following structures:

Preparation of compound of formula I

The compound of formula I can be prepared by the following route: starting from intermediate A and substituting R ₁ X ₁ to obtain I-1, removing tert-butoxycarbonyl and tert-butanesulfinyl with acid to obtain intermediate I-2, and finally and intermediate Form I-3 undergoes a substitution reaction to obtain a compound of formula I.

Alternatively, the compound of formula I can be prepared by the following route: starting from Intermediate A, the tert-butoxycarbonyl group is removed by trimethylsilyl trifluoromethanesulfonate to obtain 1-4, and then substituted with I-3 to obtain I-5. The benzyloxycarbonyl group is removed with sodium hydroxide aqueous solution to obtain intermediate I-6, and then it is substituted with intermediate R ₁ X ₁ to obtain I-7, and finally the tert-butylsulfinyl group is removed with acid to obtain the compound of formula I.

Wherein, the R ₁ , R ₂ , R ₃ , X and Y are as described above; X ₁ and X ₂ are halogen.

Pharmacology and uses

Src Hommolgy-2 Phosphatase (SHP2) is a protein tyrosine phosphatase encoded by the PTPN11 gene, which promotes a variety of cell functions, including proliferation, differentiation, cell cycle maintenance and migration. SHP2 is involved in signaling via the Ras-mitogen-activated protein kinase, JAK-STAT, or phosphoinositide 3-kinase-AKT pathway. SHP2 mediates the activation of receptor tyrosine kinases such as ErbB1, ErbB2 and c-Met's Erk1 and Erk2MAP kinases.

SHP2 has two N-terminal Src homolgy2 domains (N-SH2 and C-SH2), a catalytic domain (PTP) and a C-terminal tail. The two SH2 domains control the subcellular localization and functional regulation of SHP2. The molecule exists in an inactive conformation and inhibits its own activity via a binding network involving residues from the N-SH2 and PTP domains. In response to growth factor stimulation, SHP2 binds to specific tyrosine-phosphorylation sites on docking proteins such as Gab1 and Gab2 via its SH2 domain. This caused a conformational change, which resulted in SHP2 activation.

Mutations in PTPN11 have been identified in a variety of human diseases, such as Noonan syndrome, Leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid Leukemia and cancer of the breast, lung, and colon. SHP2 is an important downstream signaling molecule for a variety of receptor tyrosine kinases, including platelet-derived growth factor (PDGF-R), fibroblast growth factor (FGF-R) and epidermal growth factor (EGF-R) receptors. SHP2 is also an important downstream signaling molecule that activates the mitogen-activated protein (MAP) kinase pathway, which can lead to cell transformation (a necessary condition for cancer development). Knockdown of SHP2 significantly inhibited the growth of lung cancer cell lines with SHP2 mutations or EML4/ALK translocation and EGFR-amplified breast cancer and esophageal cancer. SHP2 is also the downstream activation of oncogenes in gastric cancer, anaplastic large cell lymphoma and glioblastoma.

Noonan Syndrome (NS) and Leopard Spot Syndrome (LS)-PTPNll mutations cause LS (multiple pigmented nevus syndrome, abnormal electrocardiogram conduction, distance between the eyes, pulmonary valve stenosis, abnormal genitalia, growth retardation, Sensorineural deafness) and NS (including heart defects, craniofacial deformities, and congenital abnormalities of short stature). These two disorders are part of a family of autosomal dominant syndromes caused by germline mutations in components of the RAS/RAF/MEK/ERK mitogen-activated protein kinase pathway (required for normal cell growth and differentiation). Abnormal regulation of this pathway has a profound impact, especially on the development of the heart, leading to a variety of abnormalities, including valvuloseptal defects and/or hypertrophic cardiomyopathy (HCM). It has been determined that the disturbance of the MAPK signaling pathway is important for these disorders, and some candidate genes along this pathway have been identified in humans, including KRAS, NRAS, SOS1, RAF1, BRAF, MEK1, MEK2, SHOC2 and Mutations in CBL. The most frequently mutated gene in NS and LS is PTPN11. Germline mutations of PTPN11 (SHP2) are found in ~50% of NS cases and almost all LS patients with certain characteristics of NS. For NS, Y62D and Y63C in the protein are the most common mutations. These two mutations affect the catalytically inactive conformation of SHP2 without interfering with the binding of phosphatase to its phosphorylation signal partner.

Juvenile myelomonocytic leukemia (JMML)—Somatic mutations in PTPN11 (SHP2) occur in approximately 35% of JMML (a childhood myelodysplastic (MPD)) patients. These gain-of-function mutations are usually point mutations in the N-SH2 domain or the phosphatase domain, which prevent self-inhibition between the catalytic domain and the N-SH2 domain, resulting in SHP2 activity.

Acute Myeloid Leukemia-Already in ~10% of pediatric acute leukemias such as myelodysplastic syndrome (MDS), ~7% of B-cell acute lymphoblastic leukemia (B-ALL) and ~4% of acute myeloid leukemia PTPN11 mutation was identified in (AML).

Mutations in NS and leukemia cause changes in amino acids at the interface formed by the N-SH2 and PTP domains in the self-inhibiting SHP2 conformation, disrupting inhibitory intramolecular interactions, leading to excessive activity in the catalytic domain.

SHP2 acts as a positive regulator in receptor tyrosine kinase (RTK) signaling. Cancers containing RTK changes (EGFR ^amp , Her2 ^amp , FGFR ^amp , Met ^amp , translocation/activated RTK, namely ALK, BCR/ABL) include esophageal cancer, breast cancer, lung cancer, colon cancer, gastric cancer, glioma, Head and neck cancer.

Esophageal cancer (or esophageal cancer) is a malignant disease of the esophagus. There are many subtypes, mainly squamous cell carcinoma (<50%) and adenocarcinoma. There is a high rate of RTK expression in esophageal adenocarcinoma and squamous cell carcinoma. Therefore, the SHP2 inhibitor of the present invention can be used in innovative treatment strategies.

Breast cancer is an important type of cancer and the leading cause of death for women, where patients develop resistance to existing drugs. There are four main subtypes of breast cancer, including luminal A, luminal B, Her21ik, and three-negative/Basal-like. Triple negative breast cancer (TNBC) is an aggressive breast cancer lacking specific targeted therapy. Epidermal growth factor receptor I (EGFR) has emerged as a promising target in TNBC. Inhibition of Her2 and EGFR via SHP2 may be a promising treatment for breast cancer.

Lung cancer-NSCLC is currently an important cause of cancer-related mortality. It accounts for about 85% of lung cancers (mainly adenocarcinoma and squamous cell carcinoma). Although cytotoxic chemotherapy is still an important part of treatment, targeted therapy based on genetic changes in tumors such as EGFR and ALK is more likely to benefit from targeted therapy.

Colon cancer-Approximately 30% to 50% of colorectal tumors are known to have mutated (abnormal) KRAS, and BRAF mutations occur in 10 to 15% of colorectal cancers. For the subgroup of patients whose colorectal tumors have been shown to overexpress EGFR, these patients present a favorable clinical response to anti-EGFR therapy.

Gastric cancer is one of the most popular types of cancer. Abnormal expression of tyrosine kinases (as reflected by abnormal tyrosine phosphorylation in gastric cancer cells) is known in the art. Three receptor tyrosine kinases are often amplified in gastric cancer, namely c-met (HGF receptor), FGF receptor 2 and erbB2/neu. Therefore, the destruction of different signal pathways can promote the progression of different types of gastric cancer.

Neuroblastoma is a pediatric tumor of the developing sympathetic nervous system, accounting for about 8% of childhood cancers. Genomic changes in the anaplastic lymphoma kinase (ALK) gene have been proposed to promote the pathogenesis of neuroblastoma.

Squamous cell carcinoma of the head and neck (SCCHN)-High levels of EGFR expression are associated with poor prognosis and resistance to radiation therapy in many cancers, most commonly squamous cell carcinoma of the head and neck (SCCHN). Blocking of EGFR signal leads to inhibition of receptor stimulation, cell proliferation, invasion and metastasis decline. Therefore, in SCCHN, EGFR is the best target for new anti-cancer therapies.

The present invention relates to compounds capable of inhibiting the activity of SHP2. The present invention also provides a preparation method of the compound of the present invention and a pharmaceutical preparation containing the compound. Another aspect of the present invention relates to a method of treating a disease or condition mediated by SHP2-, which method comprises the step of administering a therapeutically effective amount of a compound of formula I according to the present invention to a patient in need thereof.

In some embodiments, the present invention relates to the method as described above, wherein the SHP2-mediated disease or disorder is selected from but not limited to the following cancers: JMML, AML, MDS, B-ALL, neuroblasts Tumor, esophageal cancer, breast cancer, lung cancer, colon cancer, stomach cancer, head and neck cancer.

The compounds of the present invention can also be used to treat other diseases or disorders related to the abnormal activity of SHP2. Therefore, as a preferred embodiment, the present invention relates to a method for treating diseases or conditions selected from the group consisting of: NS, LS, JMML, AML, MDS, B-ALL, neuroblastoma, esophageal cancer, breast cancer, lung cancer, colon cancer , Stomach cancer, head and neck cancer.

The SHP2 inhibitor of the present invention can be combined with other pharmacologically active compounds or with two or more other pharmacologically active compounds, especially in the treatment of cancer. For example, the compound of formula (I) according to the present invention or a pharmaceutically acceptable salt thereof can be administered simultaneously, sequentially or separately in combination with one or more substances selected from the group consisting of chemotherapeutic agents, such as mitotic inhibitors, such as Taxanes, vinca alkaloids, paclitaxel, docetaxel, vinblastine, vinblastine, vinorelbine or vinflunine, other anticancer agents such as cisplatin, 5-fluorouracil or 5-fluoro-2 -4(1H,3H)-pyrimidinedione (5FU), flutamide or gemcitabine.

Some combinations can provide significant advantages in therapy, including synergistic activity.

In some embodiments, the present invention relates to a method as described above, wherein the compound is administered parenterally.

In some embodiments, the present invention relates to a method as described above, wherein the compound is administered intramuscularly, intravenously, subcutaneously, orally, pulmonary, intrathecal, topically, or intranasally.

In some embodiments, the present invention relates to a method as described above, wherein the compound is administered systemically.

In some embodiments, the present invention relates to a method as described above, wherein the patient is a mammal.

In some embodiments, the present invention relates to a method as described above, wherein the patient is a primate.

In some embodiments, the present invention relates to a method as described above, wherein the patient is a human.

In some embodiments, the present invention relates to a method of treating SHP2-mediated diseases or conditions, the method comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of a chemotherapeutic agent and a therapeutically effective amount of a chemotherapeutic agent as described in the present invention. Combinations of compounds of formula I.

The main advantages of the present invention include:

1. A compound of formula I is provided.

2. A novel structure of SHP2 inhibitor and its preparation and application are provided, and the inhibitor has high inhibitory activity on SHP2.

3. A pharmaceutical composition for treating SHP2 related diseases or disorders is provided.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples usually follow the conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

The starting materials used in the following examples can be purchased from chemical vendors such as Aldrich, TCI, Alfa Aesar, Beide, Anaiji, etc., or can be synthesized by known methods.

In the following examples, ice bath refers to -5°C to 0°C, room temperature refers to 10°C to 30°C, and reflux temperature generally refers to the reflux temperature of the solvent under normal pressure. The reaction overnight generally means 8-15 hours. In the following examples, where the specific operating temperature is not limited, all are performed at room temperature.

In the following examples, the separation and purification of the intermediate and the final product is by normal phase or reverse phase chromatographic column separation or other suitable methods. The normal phase flash chromatography column uses ethyl acetate and n-hexane or methanol and dichloromethane as the mobile phase. Reversed-phase preparative high pressure liquid chromatography (HPLC) uses C18 column and UV 214nm and 254nm to detect, and its mobile phase is A (water and 0.1% formic acid), B (acetonitrile) or mobile phase A (water and 0.1% carbonic acid) Hydrogen ammonium), B (acetonitrile).

In each embodiment: LCMS instrument: Pump Agilent 1260 UV detector: Agilent 1260 DAD Mass Spectrometer API 3000;

Chromatography column: Waters sunfire C18, 4.6×50mm, 5um;

Mobile phase: A-H2O (0.1% HCOOH); B-acetonitrile NMR;

Instrument: Bruker Ascend 400M (1H NMR: 400 MHz; ¹³ C NMR: 100 MHz).

Synthesis of Intermediate A: 2-Phenyl 8-(tert-butyl)(S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5] Decane-2,8-Dicarboxylate

Step 1: Add 1-(tert-butyl)4-ethylpiperidine-1,4-dicarboxylate (20.0g, 77.7mmol), THF (200mL) to a dry 1L three-necked flask under the protection of nitrogen. , LDA (2.0M n-hexane solution, 46.6 mL, 93.2 mmol) was added dropwise at -78°C, and the reaction solution was stirred at -40°C for 2 hours. Use a needle to introduce the reaction solution into another three-necked flask containing chloroacetyl chloride (50.0g, 442.9mmol) in THF (200mL) at -60°C under the protection of nitrogen. The reaction solution is slowly raised to room temperature under stirring. , The resulting white suspension was stirred at room temperature for 2 hours. Saturated sodium bicarbonate solution and ethyl acetate were added, the organic phase was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (0 to 10% ethyl acetate/petroleum ether gradient) to give an impure yellow oil A-1 (34.0 g). There is no further purification directly into the next reaction.

Step 2: Add compound A-1 (34.0 g), DMF (300 mL) and sodium azide (5.56 g, 85.5 mmol) to a dry 500 mL single-neck flask in sequence. The reaction solution was stirred at 25°C for 3 hours. The reaction solution was diluted with water, and the aqueous phase was extracted 3 times with a mixture of petroleum ether: ethyl acetate (1:1). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oil A-2 (34.6 g, crude product). There is no further purification directly into the next reaction.

Step 3: Add compound A-2 (34.6 g, crude product), THF (300 mL), H ₂ O (30 mL) and triphenylphosphine (26.5 g, 101.0 mmol) into a dry 500 mL single-neck flask in sequence. The reaction solution was stirred at 60°C for 4 hours. After cooling to room temperature, the reaction solution was diluted with water and extracted twice with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/ethyl acetate=4:1) to obtain yellow solid A-3 (8.0 g, yield: 38.5%).

¹ H NMR (400MHz, CDCl ₃ ) δ6.54 (brs, 1H), 3.91 (s, 2H), 3.85-3.75 (m, 2H), 3.65-3.55 (m, 2H), 1.89-1.76 (m, 2H) ), 1.74–1.64 (m, 2H), 1.47 (s, 9H).

Step 4: Add compound A-3 (4.0 g, 14.9 mmol), THF (40 mL) and sodium borohydride (282 mg, 7.45 mmol) to a dry 100 mL single-neck flask in sequence. The reaction solution was stirred at 25°C for 1 hour. After the reaction is completed, the reaction liquid is filtered. The obtained filtrate (the THF solution of compound A-4) was directly used in the next reaction.

Step 5: Add the THF solution (40mL) of compound A-4 obtained in the previous step and borane dimethyl sulfide (22.4mL, 44.7mmol, 2M tetrahydrofuran solution) into a dry 100mL single-neck flask with stirring at -30℃. . The reaction solution was slowly raised to 25°C within 1 hour under stirring, and then stirred at 60°C for 6 hours. After the reaction was completed, methanol (10 mL) was added dropwise to the reaction solution for quenching, and the resulting mixture was concentrated under reduced pressure to remove volatiles. The obtained residue was dissolved in methanol (20 mL), and the obtained solution was stirred at 70°C for 4 hours. The solution was concentrated under reduced pressure, and the obtained residue was purified by silica gel chromatography (dichloromethane/methanol=10:1-1:1) to obtain white solid A-5 (1.35 g, yield: 35.3%).

¹ H NMR (400MHz, CDCl ₃ ) δ 3.96-3.90 (m, 1H), 3.70-3.56 (m, 2H), 3.30--3.10 (m, 3H), 3.00-2.83 (m, 3H), 1.80-1.70 (m,1H),1.60–1.36(m,12H);

LCMS: m/z 257.2 [M+H] ⁺ .

Step 6: Add compound A-5 (1.35g, 5.27mmol), THF (14mL), water (7mL), sodium bicarbonate (2.22g, 26.4mmol) and benzyl chloroformate ( 1.35g, 7.91mmol). The reaction solution was stirred at 20°C for 12 hours. After the reaction was completed, the reaction solution was diluted with water (60 mL), and extracted twice with ethyl acetate (60 mL). The combined organic phase was washed twice with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:1) to obtain A-6 (1.40 g, yield: 68.0%) as a colorless oil.

¹ H NMR (400MHz, CDCl ₃ ) δ 7.39-7.29 (m, 5H), 5.13 (s, 2H), 4.00-3.90 (m, 1H), 3.85-3.65 (m, 3H), 3.55--3.30 (m ,3H), 3.20–2.90(m,2H), 1.78–1.53(m,4H), 1.52-1.42(m,10H), 1.40–1.32(m,1H);

LCMS: m/z 413.0 [M+Na] ⁺ .

Step 7: Add compound A-6 (2.30 g, 5.89 mmol), DCM (50 mL) and Dess-Martin oxidant (3.75 g, 8.83 mmol) into a dry 100 mL single-neck flask in sequence. The reaction solution was stirred at 25°C for 1 hour. After the completion of the reaction detected by the TLC plate, saturated sodium bicarbonate solution (30 mL) and saturated sodium thiosulfate solution (30 mL) were added to the reaction solution, and the resulting mixture was stirred at 25° C. for 30 minutes. Extract twice with ethyl acetate (60 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:0 to 2:1) to obtain colorless oil A-7 (2.10 g, yield: 91.7%).

¹ H NMR (400MHz, CDCl ₃ ) δ7.42--7.30 (m, 5H), 5.18 (s, 2H), 4.00 - 3.80 (m, 4H), 3.77 - 3.68 (m, 2H), 3.12 - 2.96 (m ,2H), 1.80–1.70(m,2H), 1.53–1.40(m,11H).

Step 8: Add compound A-7 (1.60g, 4.12mmol), (R)-tert-butylsulfinamide (999mg, 8.24mmol), THF (30mL) and tetraethyl titanate in sequence in a dry 100mL single-mouth flask Esters (2.83 g, 12.4 mmol). The reaction solution was stirred at 20°C for 40 hours. After the reaction was completed, ethyl acetate (30 mL) and saturated sodium chloride solution (6 mL) were added to the reaction solution under stirring, and the resulting mixture was filtered. The filtrate was extracted 3 times with ethyl acetate (40 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:0 to 2:1) to obtain white solid A-8 (1.60 g, yield: 78.8%).

¹ H NMR (400MHz, CDCl ₃ ) δ7.41--7.28 (m, 5H), 5.20-5.10 (m, 2H), 4.77-4.67 (m, 1H), 4.49-4, 39 (m, 1H), 4.10 --3.90(m,2H),3.68–3.55(m,1H),3.53–3.46(m,1H),3.02–2.88(m,2H),1.88–1.77(m,1H),1.76–1.64(m, 2H), 1.63-1.43 (m, 11H), 1.26 (s, 9H).

Step 9: Add compound A-8 (500mg, 1.02mmol) in THF (10mL) solution and diisobutylaluminum hydride solution (0.89mL, 1.33mmol) to a dry 50mL three-necked flask under nitrogen protection at -78℃. 1.5M). The reaction solution was stirred at -78°C for 0.5 hour. After the reaction was completed, the reaction was quenched with potassium sodium tartrate (20 mL), and stirring was continued for 0.5 hour at 20°C. The resulting mixture was extracted 3 times with ethyl acetate (30 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:0-1:1) to obtain A-9 (300 mg, yield: 59.5%) as a colorless oil.

Step 10: Add compound A-9 (300 mg, 0.608 mmol), ethanol (3 mL) and 10% aqueous sodium hydroxide solution (3 mL) in a dry 25 mL single-neck flask in sequence. The reaction solution was stirred at 85°C for 8 hours. After cooling to room temperature, the reaction solution was adjusted to pH=7 with 0.5N dilute hydrochloric acid, and the resulting mixture was concentrated under reduced pressure to remove volatiles. A mixed solvent (10 mL) of DCM/MeOH (2:1) was added to the obtained residue, filtered, and the obtained filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=1:0-10:1) to obtain a colorless oil A (210 mg, yield: 95.9%).

Synthesis of intermediate B1: (R)-N-((S)-8-(5-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2, 8-diazaspiro[4.5]decane-4-yl)-2-methylpropyl-2-sulfinamide

Step 1: Add compound A-9 (330mg, 0.668mmol) and DCM (10mL) in a dry 25mL single-mouth flask, add TMSOTf (298mg, 1.34mmol) dropwise at 0℃, and stir the reaction solution at 20℃2 hour. LCMS detected that the reaction was complete. The reaction solution was quenched by adding methanol (1 mL) and saturated sodium bicarbonate solution (1 mL). The resulting mixture was diluted with water (20 mL) and extracted twice with dichloromethane (20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain B1-1 (263 mg, crude product) as a colorless oil.

LCMS: m/z 394.1 [M+H] ⁺ .

Step 2: Add compound B1-1 (263mg, 0.668mmol), acetonitrile (5.0mL), N,N-diisopropylethylamine (1.0mL) and 3-chloro-4-into a dry 25mL single-mouth flask in sequence ((5-Chloropyrazine-2-yl)thio)pyridin-2-amine (C2, 182 mg, 0.668 mmol). The reaction solution was stirred at 90°C for 10 hours. The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=10:1) to obtain a yellow solid B1-2 (400 mg, yield: 95.0%).

LCMS: m/z 630.0 [M+H] ⁺ .

Step 3: Add compound B1-2 (400 mg, 0.635 mmol), ethanol (9 mL) and 10% aqueous sodium hydroxide solution (3 mL) to a dry 25 mL single-necked flask in sequence. The reaction solution was stirred at 85°C for 8 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure to remove volatiles. The obtained residue was diluted with water (20 mL), and extracted 3 times with a mixed solvent of dichloromethane and methanol (10/1, 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid B1 (300 mg, crude product).

LCMS: m/z 496.1 [M+H] ⁺ .

Synthesis of intermediate B2: 5-chloro-8-iodo-[1,2,4]triazolo[4,3-c]pyrimidine

Step 1: Add 2,4-dichloro-5-iodopyrimidine (1.096g, 4mmol) and 20mL of absolute ethanol to a dry 100mL flask. Under the condition of nitrogen at 0°C, 80% hydrazine hydrate (601 mg, 12 mmol) was slowly added to the mixture to continue stirring and reacting for 1 hour. After the reaction was completed, the mixture was filtered and washed with absolute ethanol to obtain 2-chloro-4-hydrazino-5-iodopyrimidine B2-1 (850 mg, yield 78.7%).

¹ H NMR (400MHz, CDCl3) δ 8.29 (s, 1H), 6.67 (s, 1H), 4.08 (s, 2H);

LCMS: m/z 271.1 [M+H] ⁺ .

Step 2: Add 2-chloro-4-hydrazino-5-iodopyrimidine (810 mg, 3 mmol) and trimethyl orthoformate (10 mL) to a dry 100 mL flask in sequence. Under nitrogen, the mixture was heated to 85°C and stirred for 5 hours. After the completion of the reaction, the residue obtained was poured into saturated NaCl solution (50 mL), and extracted with ethyl acetate (3×30 mL), washed with saturated brine and mixed with the organic layer, dried over anhydrous sodium sulfate, filtered and reduced The residue obtained by pressure concentration was purified by silica gel chromatography (0 to 50% ethyl acetate/petroleum ether) to give 5-chloro-8-iodo-[1,2,4]triazolo[4 ,3-c]pyrimidine B2 (420 mg, yield: 50%).

LCMS: m/z 280.9 [M+H] ⁺ .

Synthesis of intermediate B3: 5-chloro-8-iodoimidazo[1,2-c]pyrimidine

Step 1: Add 2,4-dichloro-5-iodopyrimidine (1.37g, 5mmol) and 2,2-dimethoxyethylamine (8.4g, 10mmol) and absolute ethanol ( 50mL). Then, under nitrogen at 0°C, triethylamine (1.01 g, 10 mmol) was slowly added dropwise to the reaction mixture, and then the mixture was stirred and reacted at room temperature for 10 hours. After the reaction is completed, it is concentrated in vacuo, the obtained concentrate is added with 15 mL of water, and extracted with dichloromethane (3×50 mL), washed with saturated brine and mixed with the organic layer, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a white solid 2-chloro-N-(2,2-dimethoxyethyl)-5-iodopyrimidin-4-amine (B3-1, 1.46 g, yield: 85%).

LC-MS: m/z 344.2 [M+H] ⁺ .

Step 2: Add 2-chloro-N-(2,2-dimethoxyethyl)-5-iodopyrimidin-4-amine (1.03g, 3mmol) and 10mL concentrated sulfuric acid to a dry 100mL flask in sequence. Under nitrogen, the mixture was heated to 65°C and stirred for 2 hours. After the reaction is complete, the reaction solution is cooled to room temperature, the mixture is slowly poured into ice water, and then the pH is adjusted to about 6-7 with 4M NaOH solution, and the off-white solid 8-iodoimidazo[1,2-c]pyrimidine is obtained by filtration. 5-ol (B3-2, 0.407 g, yield 52%).

1H NMR(400MHz,DMSO-d6)δ11.81(s,1H),7.93(d,J=1.4Hz,1H),7.60(s,1H), 7.40(d,J=1.4Hz,1H);

LC-MS: m/z 262.2 [M+H] ⁺ .

Step 3: Add 8-iodoimidazo[1,2-c]pyrimidin-5-ol (0.522g, 2mmol) and phosphorus oxychloride (8mL) into a dry 50mL single-neck flask in sequence, slowly under the protection of nitrogen N,N-diisopropylethylamine (1 mL) was added dropwise, and then the mixture was heated to 120°C and stirred for 5 hours. After the completion of the reaction, the reaction solution was cooled to room temperature and concentrated in vacuo, and then quenched by adding saturated sodium bicarbonate solution, extracted with ethyl acetate (3×40 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue obtained The product was purified by silica gel chromatography (0 to 30% ethyl acetate: petroleum ether gradient) to give a pale yellow solid 5-chloro-8-iodoimidazo[1,2-c]pyrimidine (B3, 0.360g, yield: 55%).

¹ H NMR(400MHz,DMSO-d6)δ8.24(s,1H), 8.20(d,J=1.4Hz,1H), 7.81(d,J=1.4Hz,1H);

LC-MS: m/z 280.1 [M+H] ⁺ .

Synthesis of intermediate B4: 5-chloro-7-methyl-8-iodoimidazo[1,2-c]pyrimidine

Step 1: Add ammonia (25-28% solution, 10 mL) to 2,4-dichloro-6-methylpyrimidine (8.0 g, 49.08 mmol) in EtOH (60 mL) solution under ice water bath. The reaction was stirred at room temperature for 16 h, and ammonia water (25-28% solution, 5 mL) was added. The reaction was stirred at room temperature for 8 hours, and ammonia water (25-28% solution, 5 mL) was added. Stir at room temperature for 16h. After the reaction solution was concentrated to dryness under reduced pressure, the residue obtained was purified by silica gel chromatography (0 to 50% gradient ethyl acetate: petroleum ether) to obtain a pale yellow solid 2-chloro-6-methylpyrimidin-4-amine ( B4-1, 2.4g, yield: 34%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.25 (s, 2H), 6.22 (s, 1H), 2.18 (s, 3H).

Step 2: Add NIS (3.29 g, 14.63 mmol) to a DMF (15 mL) solution of 2-chloro-6-methylpyrimidin-4-amine (B4-1, 1.4 g, 9.75 mmol). The reaction was stirred at 30°C for 16h. The reaction solution was poured into water, filtered, the filter cake was washed with saturated sodium sulfite solution, and washed with water to obtain a pale yellow solid 2-chloro-5-iodo-6-methylpyrimidin-4-amine (B4-2, 350mg, yield: 13% ).

¹ H NMR(400MHz,DMSO-d ₆ )δ7.25(s,2H), 6.22(s,1H), 2.18(s,3H);

LC-MS: m/z 269.9 [M+H] ⁺ .

Step 3: Dissolve 2-chloro-5-iodo-6-methylpyrimidin-4-amine (B4-2, 350 mg, 1.30 mmol) in an aqueous solution of chloroacetaldehyde (40 wt.% in H ₂ O, 5 mL). Heat to 100°C and stir for 2h. The reaction liquid was cooled to room temperature, filtered, and the filter cake was washed with water to obtain a pale yellow solid 8-iodo-7-methylimidazo[1,2-c]pyrimidin-5-ol (B4-3, 330mg, yield: 67%) .

¹ H NMR (400MHz, DMSO-d ₆ ): δ 12.48 (br, 1H), 8.05 (s, 1H), 7.59 (s, 1H), 2.46 (s, 3H).

Step 4: Add DIPEA to 8-iodo-7-methylimidazo[1,2-c]pyrimidin-5-ol (B4-3, 320mg, 1.16mmol) dissolved in phosphorus oxychloride (10mL) (226mg, 1.75mmol). Heat to 100°C for 6h. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and EtOAc (30 mL) was added. Wash with saturated aqueous sodium bicarbonate (3×30 mL), saturated brine (3×30 mL), dry with anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness under reduced pressure. The residue obtained is passed through silica gel chromatography (0 to 10%). Gradient methanol: dichloromethane) was purified to obtain 5-chloro-8-iodo-7-methylimidazo[1,2-c]pyrimidine (B4, 135 mg, yield: 40%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 8.16 (d, J = 1.2 Hz, 1H), 7.74 (d, J = 1.2 Hz, 1H), 2.66 (s, 3H).

Synthesis of Intermediate B5: 7-chloro-8-iodo-5-(methylsulfoxy)imidazo[1,2-c]pyrimidine

Step 1: Add NIS (12.3g, 54.66mmol) to a solution of 4-amino-6-chloro-2-methylthiopyrimidine (8.0g, 45.55mmol) in dichloromethane (150mL) under ice water bath. The reaction was stirred at 30°C for 5 hours, and the reaction was completed. The reaction solution was concentrated under reduced pressure to dryness, then methanol (50mL) was added, poured into saturated sodium sulfite aqueous solution (150mL), filtered, and the filter cake was washed with water to obtain a pale yellow solid 4-amino-6-chloro-5-iodo-2-methylthio Pyrimidine (B5-1, 12.8 g, yield: 93%).

LC-MS: m/z 301.9[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δ 7.98 (br, 1H), 6.92 (br, 1H), 2.41 (s, 3H).

Step 2: Add 4-amino-6-chloro-5-iodo-2-methylthiopyrimidine (2.0 g, 6.63 mmol) to a 40 wt.% aqueous solution of chloroacetaldehyde (20 mL). Heat to 100°C for 3 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by silica gel chromatography (0 to 10% gradient methanol: dichloromethane) and washed with ethyl acetate to obtain a yellow solid, 7-chloro-8-iodo-5-(methylthio)imidazo [1,2-c] Pyrimidine (B5-2, 1.9 g, yield: 88%).

LCMS: m/z 325.8[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δ 8.06 (d, J = 1.2 Hz, 1H), 7.73 (d, J = 1.2 Hz, 1H), 2.76 (s, 3H).

Step 3: Add 7-chloro-8-iodo-5-(methylthio)imidazo[1,2-c]pyrimidine (B5-2, 1.20g, 3.69mmol) in dichloromethane (20mL) under ice water bath M-CPBA (1.27g, 7.37mmol) was added to the solution. The reaction was stirred at 25°C for 16 hours. The reaction was quenched with ice water, washed with saturated sodium bicarbonate aqueous solution (3×20 mL), saturated brine (3×20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. Silica gel chromatography (0 to 10%) Gradient methanol: dichloromethane) and eluted with ethyl acetate to give a yellow solid 7-chloro-8-iodo-5-(methylsulfinyl)imidazo[1,2-c]pyrimidine (B5, 550mg, yield Rate: 44%).

LCMS: m/z 341.8[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δ 8.64 (d, J = 1.2 Hz, 1H), 7.85 (d, J = 1.2 Hz, 1H), 3.12 (s, 3H).

Synthesis of Intermediate C1: Sodium 2-amino-3chloropyrimidine-4sulfur

Step 1: Add 3-chloro-4-iodopyridin-2-amine (2.5g, 9.82mmol, 1.0eq), 4,5-bisdiphenylphosphine-9, to a dry 100mL round bottom three-necked flask in sequence 9-dimethylxanthene (341mg, 0.59mmol, 0.06eq), palladium acetate (110mg, 0.49mmol, 0.05eq), DIPEA (3.25mL, 19.6mmol, 2.0q), methyl 3-mercaptopropionate ( 1.19mL, 10.8mmol, 1.1eq) and 1,4-dioxane (32.5mL). Under stirring, replace with nitrogen three times, then heat to 100°C and react for 3 hours. After the completion of the reaction, the reaction solution was cooled to room temperature, diluted with ethyl acetate (50mL) and filtered under reduced pressure. The filter cake was washed with ethyl acetate (25mL), the obtained filtrate was concentrated in vacuo, and the residue obtained was chromatographed on silica gel. Purification by method (0 to 30% ethyl acetate: petroleum ether) to obtain a yellow solid C1-1 (2.0 g, yield: 78%).

¹ H NMR(400MHz,CDCl ₃ )δ7.89(d,J=5.4Hz,1H), 6.53(d,J=5.5Hz,1H), 4.87(s,2H), 3.74(s,3H), 3.24 (t, J=7.5 Hz, 2H), 2.75 (t, J=7.5 Hz, 2H).

Step 2: In a dry 100mL round bottom three-necked flask, dissolve C1-1 (2g, 8.11mmol, 1.0eq) in tetrahydrofuran (28mL), under nitrogen protection, add ethanol to the reaction solution dropwise at room temperature Sodium (2.9g, 8.51mmol, 1.05eq, 20%wt), then stirred for one hour. After the completion of the reaction, it was diluted with dichloromethane (60 mL) and sonicated for 5 minutes, filtered under reduced pressure, and the filter cake was dried in vacuum to obtain a yellow solid C1 (1.4 g, yield: 89%).

Synthesis of intermediate C2: 3-chloro-4-((5-chloropyrazin-2-yl)thio)pyridin-2-amine

Step 1: Add 2,5-dichloropyrazine (3g, 20.1mmol, 1.0eq), potassium carbonate (2.78g, 20.1mmol, 1.0eq), DMF (25mL) and 3 to a dry 100mL round-bottom flask. -Methyl mercaptopropionate (2.54g, 21.1mmol, 1.05eq), then stirred at 25 degrees Celsius for 18 hours. After the reaction, it was diluted with ethyl acetate (100 mL), washed twice with water (30 mL), then dried over anhydrous sodium sulfate, and filtered. The obtained filtrate was concentrated in vacuo, and the obtained residue was purified by silica gel chromatography (0 to 2.8% ethyl acetate: petroleum ether gradient) to obtain a yellow solid C2-1 (3.68 g, yield: 74%).

¹ H NMR(400MHz, CDCl ₃ )δ8.38(d,J=1.5Hz,1H), 8.22(d,J=1.5Hz,1H), 3.71(s,3H), 3.41(t,J=7.0Hz , 2H), 2.76 (t, J=7.0 Hz, 2H).

Step 2: In a dry 100mL round bottom three-necked flask, dissolve C2-1 (3.68g, 15.8mmol, 1.0eq) in tetrahydrofuran (50mL), under nitrogen protection, add dropwise to the reaction solution at room temperature Sodium ethoxide (5.65g, 16.6mmol, 1.05eq, 20%wt), then stirred for one hour. After the reaction was completed, half of the solvent was concentrated under reduced pressure, ether (200 mL) was added to the remaining reaction solution, precipitation was precipitated, and the filter cake was vacuum-dried to obtain a yellow solid C2-2 (2.5 g, yield : 84.2%).

Step 3: Under nitrogen protection, sequentially add 3-chloro-4-iodopyridin-2-amine (2g, 7.86mmol, 1.0eq) and 4,5-bis-two to a dry 100mL round bottom three-necked flask. Phenylphosphine-9,9-dimethylxanthene (363mg, 0.63mmol, 0.08eq), Pd ₂ (dba) ₃ (287mg, 0.31mmol, 0.04eq), DIPEA (2.6mL, 15.7mmol, 2.0eq) ), dioxane (48mL) and C2-2 (1.39g, 8.25mmol, 1.05eq), and then heated to 105 degrees Celsius for 14 hours. After the reaction was completed, it was cooled to room temperature, diluted with ethyl acetate, filtered under reduced pressure, and the filter cake was washed with ethyl acetate. The residue obtained by concentrating the filtrate under reduced pressure was purified by silica gel chromatography (ethyl acetate/petroleum ether with a gradient of 0 to 40%) to obtain a yellow solid C2 (1.5 g, yield: 66%).

¹ H NMR(400MHz,CDCl ₃ )δ8.45(d,J=1.4Hz,1H), 8.29(d,J=1.4Hz,1H), 7.83(d,J=5.3Hz,1H), 6.51(d , J=5.3Hz, 1H), 4.95(s, 2H).

Example 1: Synthesis of compound 1

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(methylsulfonyl)-2,8-diazepine Spiro[4.5]decane-4-amine

Step 1: Add A (60mg, 0.167mmol), dichloromethane (5mL), triethylamine (51mg, 0.501mmol) and methanesulfonic anhydride (58mg, 0.334mmol) to a 25mL round bottom flask in sequence. Stir at 20°C for 1 hour. The reaction solution was concentrated under reduced pressure, diluted with water (10 mL), and extracted 3 times with ethyl acetate (10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain colorless oil 1-1 (40 mg, yield: 54.8%).

LCMS: m/z 438.1 [M+H] ⁺ .

Step 2: Add compound 1-1 (40mg, 0.091mmol), methanol (1mL) and dioxane hydrochloride (1mL, 4.0M) to a 25mL round bottom flask in sequence, and the reaction solution was stirred at 20°C for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain yellow solid 1-2 (28 mg, hydrochloride, yield: 100%).

LCMS: m/z 234.1 [M+H] ⁺ .

Step 3: Add compound 1-2 (28mg, 0.091mmol), acetonitrile (3.0mL), N,N-diisopropylethylamine (1.0mL) and 3-chloro-4-into a dry 25mL single-neck flask in sequence ((5-Chloropyrazine-2-yl)thio)pyridin-2-amine (32 mg, 0.118 mmol). The reaction solution was stirred at 90°C for 7 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by thin-layer chromatography on a silica gel plate (dichloromethane/methanol=10:1) to obtain 30 mg of impure product. Then, a white solid 1 (17 mg, formate, yield: 27.9%) was prepared by HPLC.

1H NMR(400MHz,DMSO-d6)δ8.48(d,J=1.6Hz,1H), 8.29(d,J=1.2Hz,1H), 8.19(s,1H), 7.65(d,J=5.6Hz ,1H),6.35(s,2H),5.81(d,J=5.6Hz,1H),4.20–4.06(m,2H),3.55–3.47(m,1H),3.45–3.38(m,1H), 3.37–3.25(m,2H), 3.20–3.13(m,2H), 3.05–2.99(m,1H), 2.93(s,3H), 1.72–1.60(m,2H), 1.53–1.43(m,2H) );

LC-MS: m/z 470.0[M+H]+.

Example 2: Synthesis of Compound 2

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(pyrazin-2-yl)-2,8- Diazaspiro[4.5]decane-4-amine

Step 1: Add compound A-8 (200mg, 0.407mmol), tetrahydrofuran/water (98:2,5mL) and sodium borohydride (46mg, 1.22mmol) to a 25mL round-bottom flask at -50°C. The solution was heated to 20°C under stirring for 3 hours. The TLC dot plate reaction is complete. The reaction solution was diluted with water (20 mL), and extracted with ethyl acetate (20 mL) three times. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain A-9 (200 mg, yield: 99.5%) as a colorless oil.

Step 2: Add compound A-9 (200mg, 0.405mmol), ethanol (4mL) and 10% sodium hydroxide aqueous solution (2mL) in a dry 25mL single-necked flask in sequence. The reaction solution was stirred at 85°C for 8 hours. After cooling to room temperature, the reaction solution was adjusted to pH=7 with 0.5N dilute hydrochloric acid, and the resulting mixture was concentrated under reduced pressure to remove volatiles. A mixed solvent (10 mL) of DCM/MeOH (2:1) was added to the obtained residue, filtered, and the obtained filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain a colorless oil A (110 mg, yield: 75.3%).

LC-MS: m/z 460.2 [M+H] ⁺ .

Step 3: Add A (110mg, 0.306mmol), acetonitrile (3mL), 2-chloropyrazine (140mg, 1.22mmol) and N,N-diisopropylethylamine (1mL) to a 25mL round bottom flask in sequence The reaction solution was stirred at 90°C for 10 hours. The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain a yellow oil 2-1 (50 mg, yield: 37.3%).

¹ H NMR (400MHz, CDCl ₃ ) δ 8.06–8.02 (m, 1H), 7.88 (d, J = 1.6 Hz, 1H), 7.84 (d, J = 2.4 Hz, 1H), 4.20-3.93 (m, 2H), 3.90--3.88 (m, 1H), 3.80 - 3.60 (m, 2H), 3.55 - 3.30 (m, 3H), 3.04 - 2.84 (m, 2H), 1.90 - 1.75 (m, 1H), 1.72-- 1.58 (m, 1H), 1.55-1.40 (m, 11H), 1.21 (s, 9H).

Step 4: Add compound 2-1 (50 mg, 0.114 mmol), methanol (1 mL), and dioxane hydrochloride (1 mL, 4.0 M) to a 25 mL round bottom flask in sequence, and the reaction solution was stirred at 20° C. for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a yellow solid 2-2 (35 mg, hydrochloride, yield: 100%).

Step 5: Add compound 2-2 (35mg, 0.114mmol), acetonitrile (3.0mL), N,N-diisopropylethylamine (1.0mL) and 3-chloro-4-into a dry 25mL single-neck flask in sequence ((5-Chloropyrazine-2-yl)thio)pyridin-2-amine (40 mg, 0.148 mmol). The reaction solution was stirred at 90°C for 10 hours. The TLC dot board shows that new dots are generated. The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=10:1) to obtain 40 mg of impure product. Then, a white solid 2 (22 mg, formate, yield: 40.7%) was obtained by HPLC preparation.

¹ H NMR(400MHz,DMSO-d6)δ8.49(d,J=1.2Hz,1H), 8.30(d,J=1.6Hz,1H), 8.24(s,1H), 8.03(dd,J=2.8 ,1.6Hz,1H),7.98(d,J=1.2Hz,1H),7.78(d,J=2.4Hz,1H),7.66(d,J=5.6Hz,1H),6.35(s,2H), 5.82(d,J=5.6Hz,1H), 4.28–4.17(m,2H), 3.80–3.72(m,1H), 3.71–3.64(m,1H), 3.50–3.43(m,1H),3.42– 3.24(m,4H), 1.80–1.65(m,2H), 1.58–1.46(m,2H);

LC-MS: m/z 470.0 [M+H] ⁺ .

Example 3: Synthesis of Compound 3.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-phenyl-2,8-diazaspiro[4.5 ]Decane-4-amine

Step 1: Add compound A (120mg, 0.334mmol), iodobenzene (136mg, 0.668mmol), dioxane (12mL), Pd ₂ (dba) ₃ (30mg, 0.033mmol) to a 25mL round bottom flask in sequence , 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (39mg, 0.067mmol), and cesium carbonate (326mg, 1.00mmol), the reaction solution was stirred at 100°C under nitrogen protection for 5 hours. The reaction solution was diluted with water (30 mL), and extracted twice with ethyl acetate (30 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:1) to obtain a yellow solid 3-1 (90 mg, yield: 62.1%).

LC-MS: m/z 436.1 [M+H] ⁺ .

Step 2: Add compound 3-1 (45mg, 0.103mmol), methanol (2mL) and dioxane hydrochloride (1mL, 4.0M) to a 25mL round bottom flask in sequence, and the reaction solution was stirred at 20°C for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a yellow solid 3-2 (31 mg, hydrochloride, yield: 100%).

LC-MS: m/z 232.1 [M+H] ⁺ .

Step 3: Add compound 3-2 (31mg, 0.103mmol), acetonitrile (3.0mL), N,N-diisopropylethylamine (1.0mL) and 3-chloro-4-into a dry 25mL single-neck flask in sequence ((5-Chloropyrazine-2-yl)thio)pyridin-2-amine (C2, 37 mg, 0.134 mmol). The reaction solution was stirred at 90°C for 7 hours. The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=5:1) to obtain 30 mg of impure product. Then, a white solid 3 (23 mg, formate, yield: 43.4%) was prepared by HPLC.

¹ H NMR(400MHz,DMSO-d6)δ8.50(s,1H), 8.30(d,J=0.8Hz,1H), 8.23(s,1H), 7.66(d,J=5.2Hz,1H), 7.16(t,J=8.0Hz,2H),6.56–6.43(m,3H),6.36(s,2H),5.81(d,J=5.2Hz,1H), 4.30–4.16(m,2H),3.59 --3.52(m,1H),3.51–3.46(m,1H), 3.35–3.24(m,3H), 3.23–3.17(m,1H), 3.11–3.03(m,1H), 1.79–1.65(m, 2H),1.54–1.44(m,2H);

LC-MS: m/z 468.0 [M+H] ⁺ .

Example Four: Synthesis of Compound 4.

(S)-1-(4-Amino-8-(5-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2,8-diaza spiro [4.5]Decan-2-yl)prop-2-en-1-one

Step 1: Add compound B (90mg, 0.181mmol), dichloromethane (2mL), and acrylic anhydride (23mg, 0.181mmol) to a round bottom flask at -30℃, and stir the reaction solution at -30℃ for 0.5 hours . After the completion of the reaction, the reaction solution was concentrated under reduced pressure at 0°C. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=10:1) to obtain a yellow solid 4-1 (43 mg, yield: 43.0%).

LC-MS: m/z 550.1 [M+H] ⁺ .

Step 2: Add compound 4-1 (43mg, 0.078mmol), methanol (1mL) and dioxane hydrochloride (0.2mL, 4.0M) to a round bottom flask in sequence, and the reaction solution was stirred at 20°C for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was prepared and purified by HPLC to obtain 4 as a white solid (21 mg, formate, yield: 55.3%).

¹ H NMR(400MHz,DMSO-d6)δ8.48(d,J=8.4Hz,1H), 8.35–8.25(m,1H), 8.23– 8.16(m,1H), 6.71–6.49(m,1H) ,6.34(d,J=8.8Hz,2H), 6.20–6.10(m,1H), 5.86–5.79(m,1H), 5.73–5.64(m,1H), 4.30–4.05(m,2H), 3.90 –3.50(m,4H), 3.40–3.10(m,4H), 1.76–1.58(m,2H), 1.52–1.36(m,2H);

LC-MS: m/z 446.1 [M+H] ⁺ .

Example 5: Synthesis of Compound 5.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(azetidin-3-yl)-2 ,8-diazaspiro[4.5]decane-4-amine

Step 1: Add compound B (120mg, 0.242mmol), 1-tert-butoxycarbonyl (Boc)-3-azetidinone (41mg, 0.242mmol), dichloromethane ( 10mL) and sodium triacetoxyborohydride (77mg, 0.363mmol), and then stirred at 25°C for 2 hours. After the reaction was completed, saturated sodium bicarbonate solution (10 mL) was added, and extraction was performed twice with dichloromethane (15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (dichloromethane/methanol=1:0-10:1) to obtain a yellow solid 5-1 (90 mg, yield: 57.0%).

LCMS: m/z 651.1 [M+H] ⁺ .

Step 2: Add compound 5-1 (20 mg, 0.031 mmol), methanol (1 mL), and dioxane hydrochloride (0.5 mL, 4.0 M) to a round bottom flask in sequence, and the reaction solution was stirred at 20° C. for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was prepared and purified by HPLC to obtain 5 as a white solid (9 mg, formate, yield: 60.0%).

¹ H NMR(400MHz,DMSO-d6)δ8.51(d,J=1.2Hz,1H), 8.31(s,1H), 8.19(s,1H), 7.66(d,J=5.2Hz,1H), 6.37(s,2H),5.80(d,J=5.6Hz,1H), 4.40–4.20(m,2H), 4.00–3.80(m,4H), 3.65–3.55(m,1H), 3.25–3.00( m,6H), 2.83–2.75(m,1H), 2.69–2.61(m,1H), 1.75–1.53(m,4H);

LCMS: m/z 447.2 [M+H] ⁺ .

Example 6: Synthesis of Compound 6.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(1-methylazacyclobutane-3- Yl)-2,8-diazaspiro[4.5]decane-4-amine

Step 1: Add compound 5-1 (60mg, 0.092mmol), dichloromethane (3mL) and TMSOTf (41mg, 0.184mmol) to a dry 10mL single-necked flask at 0℃, and then stir at 20℃ for reaction 2 hour. LCMS detected that the reaction was complete. The reaction solution was quenched by adding methanol (0.5 mL) and saturated sodium bicarbonate solution (0.5 mL). The resulting mixture was diluted with water (15 mL) and extracted 3 times with dichloromethane (15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid 6-1 (50 mg, crude product).

LCMS: m/z 551.2 [M+H] ⁺ .

Step 2: Add compound 6-1 (40mg, 0.073mmol), dichloromethane (6mL), formaldehyde (6mg, 0.073mmol, 37% aqueous solution) and water (0.1mL) to a round bottom flask at 0°C in sequence The solution and sodium triacetoxyborohydride (31mg, 0.146mmol), the reaction solution was stirred at 0°C for 0.5 hours. After the reaction was completed, saturated sodium bicarbonate solution (5 mL) was added, and extraction was performed twice with dichloromethane (6 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid 6-2 (18 mg, crude product).

LCMS: m/z 565.1 [M+H] ⁺ .

Step 3: Add compound 6-2 (18mg, 0.032mmol), methanol (1mL) and dioxane hydrochloride (0.5mL, 4.0M) to the round bottom flask in sequence, and the reaction solution was stirred at 20°C for 0.5 hours. After the completion of the reaction, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by HPLC preparation to obtain an off-white solid 6 (1 mg, formate, yield: 6.3%).

¹ H NMR(400MHz,DMSO-d6)δ8.47(d,J=1.2Hz,1H), 8.32–8.26(m,2H), 7.66(d,J=5.6Hz,1H), 6.35(s,2H ), 5.81(d,J=5.6Hz,1H), 4.34–4.12(m,2H),3.41–3.34(m,2H),3.22–3.06(m,4H),3.05–2.85(m,4H), 2.70–2.60(m,1H), 2.30–2.20(m,3H), 1.70–1.45(m,4H);

LCMS: m/z 461.1 [M+H] ⁺ .

Example 7: Synthesis of Compound 7.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-cyclobutyl-2,8-diazaspiro[ 4.5] Decane-4-amine

Referring to the synthesis method of compound 5, cyclobutanone was used instead of 1-tert-butoxycarbonyl-3-azetidinone, and compound 7 was obtained by performing a two-step reaction.

¹ H NMR(400MHz,DMSO-d6)δ8.53(d,J=1.2Hz,1H),8.32(s,1H),8.13(s,1H),7.66(d,J=5.6Hz,1H), 6.37(s,2H),5.81(d,J=5.2Hz,1H),4.50–3.50(m,6H), 3.25–3.00(m,3H), 2.45–2.05(m,4H),2.00–1.50( m, 6H), 1.18 (t, J = 7.2 Hz, 1H);

LCMS: m/z 446.2 [M+H] ⁺ .

Example 8: Synthesis of Compound 8.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(oxetan-3-yl)-2 ,8-diazaspiro[4.5]decane-4-amine

Referring to the synthesis method of compound 5, 3-oxetanone was used instead of 1-tert-butoxycarbonyl-3-azetidinone, and compound 8 was obtained by performing a two-step reaction.

¹ H NMR(400MHz,DMSO-d6)δ8.49(d,J=0.8Hz,1H), 8.28-8.15(m,2H), 7.65(d,J=5.2Hz,1H), 6.36(s,2H ), 5.80(d,J=5.2Hz,1H), 4.62–4.53(m,2H), 4.51–4.42(m,2H), 4.36–4.16(m,2H), 3.72–3.62(m,2H), 3.21--3.03(m,3H), 3.01--2.89(m,1H), 2.71--2.63(m,1H), 2.60--2.54(m,1H), 2.41--2.34(m,1H), 1.72--1.50(m ,4H);

LCMS: m/z 448.2 [M+H] ⁺ .

Example 9: Synthesis of Compound 9.

(S)-8-(8-((2-Amino-3-chloropyridin-4-yl)thio)imidazole[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8 -Diazaspiro[4.5]decane-4-amine

Step 1: Add compound 3-2 (35mg, 0.115mmol), acetonitrile (3mL), N,N-diisopropylethylamine (0.5mL) and 5-chloro-8-iodine to a dry 10mL single-necked flask. Imidazole [1,2-c] pyrimidine (B3, 32 mg, 0.115 mmol) was then stirred at 90°C for 7 hours. After the reaction solution was cooled to 20°C, Boc ₂ O (118 mg, 0.542 mmol) was added to the reaction solution, and then the reaction was stirred at 50°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the obtained residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1:1) to obtain a yellow solid 9-1 (40 mg, yield: 60.6%). CMS: m/z 575.1 [M+H] ⁺ .

Step 2: Add 2-amino-3-chloropyridine-4-sodium sulfide (14mg, 0.077mmol), compound 9-1 (40mg, 0.070mmol), dioxane (1mL), and Pd to the round bottom flask in sequence ₂ (dba) ₃ (7mg, 0.008mmol), 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (9mg, 0.016mmol) and N,N-diisopropylethylamine ( 40mg, 0.308mmol), the reaction solution was pumped with nitrogen three times, and then stirred at 100°C for 6 hours. After the reaction was completed, the reaction solution was diluted with water (5 mL) and extracted twice with ethyl acetate (5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography (ethyl acetate) to obtain a yellow solid 9-2 (15 mg, yield: 31.9%). LCMS: m/z 607.2 [M+H] ⁺ .

Step 3: Add compound 9-2 (15 mg, 0.025 mmol), dichloromethane (2 mL) and trifluoroacetic acid (0.4 mL) to a dry single-neck flask in sequence. The reaction solution was stirred at 20°C for 1 hour. The reaction solution was concentrated under reduced pressure. The resulting residue reaction solution was adjusted to pH=8 with saturated sodium bicarbonate solution, and extracted with a mixed solvent (5 mL) of DCM/MeOH (10:1) for three times. The combined organic phase was concentrated under reduced pressure to obtain an off-white solid 9 (7.3 mg, yield: 57.5%).

¹ H NMR(400MHz,DMSO-d6)δ8.03(s,1H),7.84(s,1H),7.60-7.52(m,2H),7.19-7.12(m,2H),6.60-6.50(m, 3H),6.33(brs,2H),5.79(d,J=5.6Hz,1H),3.97–3.85(m,2H),3.56–3.45(m,2H),3.43–3.35(m,2H), 3.27 –3.21(m,1H), 3.20–3.13(m,1H), 3.03–2.96(m,1H), 2.00–1.87(m,2H), 1.60–1.50(m,2H); LCMS: m/z 507.1 [M+H] ⁺ .

Example 10: Synthesis of compound 10.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(1-methyl-1H-pyrazole-3- Yl)-2,8-diazaspiro[4.5]dec-4-amine

Step 1: Add (S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (A1, 45mg, 0.125mmol) dissolved in 5mL of anhydrous 1,4-dioxane, followed by adding 3-iodo-1-methyl-1H-pyrazole (4.114mg, 0.556mmol), anhydrous potassium tert-butoxide ( 70 mg, 0.624 mmol), Brettphos Pd Gen. 3 (23 mg, 0.0254 mmol), protected by argon, heated to 100°C, and reacted for 5 hours. The reaction solution was poured into ice water, extracted with ethyl acetate 3 times, the organic phase was washed once with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and passed through a column to obtain (S)-4-(((S )-Tert-Butylsulfinyl)amino)-2-(1-methyl-1H-pyrazol-3-yl)-2,8-diazaspiro[4.5]decane-8-tert-butyl carboxy Acid (10-1, 34mg, yield: 61.84%). LCMS: m/z 437.3 [M+H] ⁺ .

Step 2: Add (S)-4-(((S)-tert-butylsulfinyl)amino)-2-(1-methyl-1H-pyrazol-3-yl)-2,8-diazepine Heterosspiro[4.5]decane-8-tert-butyl carboxylic acid (10-1, 34mg, 0.0773mmol) was dissolved in 5mL dichloromethane/methanol (1/1), under argon protection, the ice water was cooled to 0 After adding 4M HCl/1,4-dioxane (0.5 mL, 2.0 mmol), stirring at room temperature for 1 hour, the reaction was complete. The reaction solution was directly spin-dried to obtain the crude product (S)-2-(1-methyl-1H-pyrazol-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (10- 2, 0.0773mmol), directly cast the step. LCMS: m/z 236.3 [M+H] ⁺ .

Step 3: The crude product (S)-2-(1-methyl-1H-pyrazol-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (10-2, 0.0773mmol ) Dissolved in 5mL anhydrous acetonitrile, add DIPEA (0.5mL, 2.926mmol) and 3-chloro-4–((5-chloropyrazin-2-yl)thio)pyridin-2-amine (C2, 28mg, 0.1025mmol). Protected by argon, heated to 90°C and refluxed for 7 hours to complete the reaction. Cool to room temperature, spin dry, and climb the big plate to get the pure spots to climb the big plate again to obtain compound 10, (S)-8-(5—((2-amino-3-chloropyridin-4-yl)thio)pyridine Azin-2-yl)-2-(1-methyl-1H-pyrazol-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (15mg, yield: 41.18%) .

LCMS: m/z 472.5[M+H] ⁺

¹ H NMR(400MHz, MeOD) δ8.338-8.282(m,2H), 7.597(d,J=5.6Hz,1H), 7.299(d,J=2.4Hz,1H), 5.934(d,J=5.6 Hz,1H),5.521(d,J=2.4Hz,1H),4.380-4.304(m,2H),3.702(s,3H),3.621(m,1H),3.546(d,J=9.6Hz,1H ), 3.264-3.234 (m, 2H), 3,143-3.105 (m, 1H), 1.844-1.747 (m, 2H), 1.664-1.573 (m, 2H), 1.286 (s, 1H).

Example 11: Synthesis of compound 11.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(pyridin-2-yl)-2,8-bis Azaspiro[4.5]dec-4-amine

Step 1: Add (S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (A1, 100mg, 0.278mmol) dissolved in 5mL of anhydrous 1,4-dioxane, followed by adding 2-iodopyridine (114mg, 0.556mmol), anhydrous cesium carbonate (272mg, 0.835mmol), 4,5-bis-dioxane Phenylphosphine-9,9-dimethylxanthene (32mg, 0.0553mmol) and Pd ₂ (dba) ₃ (26mg, 0.0284mmol), protected by argon, heated to 100°C and reacted for 5 hours. The reaction solution was poured into ice water, extracted with ethyl acetate 3 times, the organic phase was washed once with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and passed through a column to obtain (S)-4-(((R )-Tert-butylsulfinyl)amino)-2-(pyridin-2-yl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (11-1, 33mg, Yield: 27.19%). LCMS: m/z 437.3[M+H] ⁺

Step 2: Add (S)-4-(((R)-tert-butylsulfinyl)amino)-2-(pyridin-2-yl)-2,8-diazaspiro[4.5]decane- Tert-Butyl 8-carboxylate (11-1, 33mg, 0.07558mmol) was dissolved in 5mL dichloromethane/methanol (1/1), under the protection of argon, the temperature of ice water was cooled to 0 degrees, and 4M HCl/1 was added. 4-Dioxane (0.3 mL, 1.2 mmol) was stirred at room temperature for 1 hour, and the reaction was complete. The reaction solution was spin-dried directly to obtain crude (S)-2-(pyridin-2-yl)-2,8-diazaspiro[4.5]dec-4-amine (11-2, 0.07558mmol), which was directly cast step. LCMS: m/z 304.2 [M+H] ⁺ .

Step 3: Dissolve the crude (S)-2-(pyridin-2-yl)-2,8-diazaspiro[4.5]dec-4-amine (11-2, 0.07558mmol) in 5mL anhydrous acetonitrile , DIPEA (0.5 mL, 2.926 mmol) and 3-chloro-4-((5-chloropyrazin-2-yl)thio)pyridin-2-amine (7, 27 mg, 0.09886 mmol) were added. Protected by argon, heated to 90° and refluxed for 7 hours, and the reaction was completed. Cool to room temperature, spin dry, and purify by column to obtain compound (S)-8-(5-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2- (Pyridin-2-yl)-2,8-diazaspiro[4.5]dec-4-amine (11,9.2 mg, yield: 25.95%).

LCMS: m/z 469.1 [M+H] ⁺ ;

¹ H NMR (400MHz, MeOD) δ 8.346 (d, J = 24.4 Hz, 2H), 8.085 (d, J = 5.2 Hz, 1H), 7.638-7.591 (m, 2H), 6.723-6.646 (m, 2H) ), 5.938 (d, J = 5.6Hz, 1H), 4.420-4.299 (m, 2H), 4.001-3.956 (m, 1H), 3.800-3.629 (m, 4H), 3.460-3.417 (m, 1H), 1.887-1.749 (m, 4H), 1.287 (s, 1H).

Example 12: Synthesis of compound 12.

(S)-8-(5-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-(pyridin-3-yl)-2,8-bis Azaspiro[4.5]dec-4-amine

Step 1: Add (S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (A1, 50mg, 0.139mmol) was dissolved in 5mL of anhydrous 1,4-dioxane, followed by the addition of 3-iodopyridine (4,57mg, 0.278mmol), anhydrous cesium carbonate (136mg, 0.417mmol), 4,5- Bisdiphenylphosphine-9,9-dimethylxanthene (17mg, 0.0294mmol) and Pd ₂ (dba) ₃ (13mg, 0.0298mmol), protected by argon, heated to 100 degrees, reacted for 5 hours, react complete. The reaction solution was poured into ice water, extracted with ethyl acetate 3 times, the organic phase was washed once with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and passed through a column to obtain (S)-4-(((R )-Tert-butylsulfinyl)amino)-2-(pyridin-3-yl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (12-1, 39mg, Yield: 64.26%). LCMS: m/z 437.3 [M+H] ⁺ .

Step 2: Add (S)-4-(((R)-tert-butylsulfinyl)amino)-2-(pyridin-3-yl)-2,8-diazaspiro[4.5]decane- Tert-butyl 8-carboxylate (12-1, 39mg, 0.08923mmol) was dissolved in 5mL of dichloromethane/methanol (1/1), under the protection of argon, the temperature of ice water was cooled to 0 degrees, and 4M HCl/1 was added. 4-Dioxane, then stirred at room temperature for 1 hour, the reaction was complete. The reaction solution was directly spin-dried to obtain the crude product (S)-2-(pyridin-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (12-2, 0.08923mmol), which was cast directly step. LCMS: m/z 233.5 [M+H] ⁺ .

Step 3: Dissolve the crude (S)-2-(pyridin-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (12-2, 0.08923mmol) in 5mL anhydrous acetonitrile , DIPEA (0.5 mL, 2.926 mmol) and 3-chloro-4-((5-chloropyrazin-2-yl)thio)pyridin-2-amine (7, 32 mg, 0.1172 mmol) were added. Protected by argon, heated to 90° and refluxed for 7 hours, and the reaction was completed. Cool to room temperature, spin dry, beaten with ethyl acetate, filter, and prepare a silica gel plate for purification to obtain (S)-8-(5-((2-amino-3-chloropyridin-4-yl)thio)pyrazine- 2-yl)-2-(pyridin-3-yl)-2,8-diazaspiro[4.5]dec-4-amine (compound 12, 16.4 mg, yield: 39.15%).

LCMS: m/z 469.1 [M+H] ⁺ ;

¹ H NMR (400MHz, MeOD) δ 8.382-8.318 (m, 2H), 8.036-7.960 (m, 2H), 7.601 (d, J = 5.6 Hz, 1H), 7.444-7.411 (m, 1H), 7.310 -7.284 (m, 1H), 5.965 (d, J = 5.6 Hz, 1H), 4.398-4.305 (m, 2H), 3.935-3.907 (m, 1H), 3.849-3.829 (m, 1H), 3.686-3.617 (m, 2H), 3.567-3.533 (m, 1H), 1.879-1.806 (m, 4H), 1.387-1,360 (m, 2H.

Example 13: Synthesis of Compound 13.

(S)-8-(6-Amino-5-(((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-phenyl-2,8-di Azaspiro[4.5]decyl 4-amine

Step 1: Dissolve 3-bromo-6-chloropyrazine-2-amine (208mg, 1.0mmol) and 2-amino-3-chloropyridine-4-thiol (168mg, 1.05mmol) in dioxane ( 10mL), copper iodide (95mg, 0.5mmol) and 1,10-phenanthroline (108mg, 0.6mmol) and DIPEA (260mg, 2mmol) were added. The temperature was raised to 120 degrees and reacted overnight under nitrogen. Dilute with ethyl acetate (80 mL), wash with saturated brine (100 mL), dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify by column chromatography (petroleum ether: ethyl acetate = 1:2) to obtain 3-(( 2-Amino-3-chloropyridin-4-yl)sulfanyl)-6-chloropyrazine-2-amine (13-1, 238 mg, yield: 82.6%), as a pale yellow solid. LCMS: m/z 287.9 [M+H] ⁺ .

Step 2: Combine 3-((2-amino-3-chloropyridin-4-yl)thio)-6-chloropyrazine-2-amine (13-1, 45mg, 0.15mmol) and 2-phenyl- 2,8-Diazaspiro[4.5]dec-4-amine hydrochloride (50 mg, 0.16 mmol) was dissolved in acetonitrile (10 mL), and DIPEA (180 mg, 1.47 mmol) was added. The temperature is increased to 110 degrees and the reaction is overnight. Dilute with ethyl acetate (80mL), wash with saturated brine (100mL), dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure, silica gel column chromatography (dichloromethane: methanol = 10:1) and Purp-HPLC to obtain 8 -(6-Amino-5-(((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2-phenyl-2,8-diazaspiro[4.5 ] Decane 4-amine (13, 10 mg, yield: 12.8%).

LCMS: m/z 483.5[M+H] ⁺ ;

¹ H NMR(400MHz, MeOD-d ₄ )δ7.59(t,J=2.6Hz,2H), 7.16(t,J=7.8Hz,2H), 6.63-6.57(m,3H), 5.93(d, J = 5.6Hz, 1H), 4.28-4.23 (m, 2H), 3.65-3.61 (m, 1H), 3.56-3.54 (m, 1H), 3.28-3.24 (m, 4H), 3.13-3.10 (m, 1H), 1.81-1.77 (m, 2H), 1.63-1.58 (m, 2H).

Example 14: Synthesis of compound 14.

Step 1: Combine 5-chloro-8-iodo-7-methylimidazo[1,2-c]pyrimidine (B4, 34mg, 0.12mmol) and 2-phenyl-2,8-diazaspiro[4.5 ] Deca-4-amine hydrochloride (3-2, 35 mg, 0.12 mmol) was dissolved in acetonitrile (10 mL), and DIPEA (0.2 mL, 1.98 mmol) was added. The temperature was raised to 65°C and reacted for 2h. Dilute with ethyl acetate (80 mL), wash with saturated brine (100 mL), dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify by column chromatography (dichloromethane: methanol = 30:1) to obtain a pale yellow solid 8- (8-Iodo-7-methylimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-amine (14-1 , 50mg, yield: 89.3%). LCMS: m/z 488.9 [M+H] ⁺ .

Step 2: Add 8-(8-iodo-7-methylimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4 -Amine (50mg, 0.10mmol) and sodium 2-amino-3-chloropyridine-4-sulfide (37mg, 0.2mmol) were suspended in dioxane (10mL), DIPEA (0.2mL, 1.21mmol) was added, Xantphos ( 29 mg, 0.05 mmol) and Pd ₂ (dba) ₃ (45 mg, 0.05 mmol), replaced with argon three times. The temperature was raised to 120°C and reacted for 4 hours. Diluted with ethyl acetate (80 mL), filtered, concentrated under reduced pressure, purified by column chromatography (dichloromethane: methanol = 8:1) and purified by prep-HPLC to obtain 8-(8-(((2-amino-3- (Chloropyridin-4-yl)thio)-7-methylimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec4- Amine (14, 4 mg, yield: 5.9%).

LCMS: m/z 521.2[M+H] ⁺ ;

¹ H NMR(400MHz,DMSO-d ₆ )δ8.35(s,3H),7.76(s,1H),7.55(d,J=5.2Hz,1H),7.49(s,1H),7.16(t, J = 7.6Hz, 2H), 6.57-6.52 (m, 3H), 6.31 (s, 2H), 5.71 (d, J = 5.2Hz, 1H), 3.90-3.88 (m, 2H), 3.32-3.00 (m , 7H), 2.47 (s, 3H), 1.95-1.92 (m, 2H), 1.58-1.55 (m, 2H).

Example 15: Synthesis of Compound 15.

(S)-8-(8-(((2-Amino-3-chloropyridin-4-yl)thio)-7-chloroimidazo[1,2-c]pyrimidin-5-yl)-2- Phenyl-2,8-diazaspiro[4.5]decyl 4-amine

Step 1: To 7-chloro-8-iodo-5-(methylsulfinyl)imidazo[1,2-c]pyrimidine (B5, 222mg, 0.65mmol) and (S)-2-phenyl-2 Cs ₂ CO ₃ (2.1 g, 6.45 mmol) was added to the suspension of 8-diazaspiro[4.5]decane 4-amine (3-2, 220 mg, 0.72 mmol) in acetonitrile (30 ml). The temperature was raised to 30 degrees and reacted for 1 hour. The reaction solution was diluted with water (50ml), extracted with ethyl acetate (2×30ml), the organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain a white solid (S)-8-(7- Chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec4-amine (15-1, 140mg, yield 42.4%).

LCMS: m/z 509.0 [M+H] ⁺ .

Step 2: To (S)-8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5] Triethylamine (60 mg, 0.56 mmol) and (Boc) ₂ O (120 mg, 0.56 mmol) were added to a solution of decyl 4-amine (15-1, 140 mg, 0.28 mmol) in dichloromethane (20 mL), and reacted at room temperature overnight. The reaction solution was concentrated and purified by column chromatography to obtain pale yellow solid tert-butyl (S)-(8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2- Phenyl-2,8-diazaspiro[4.5]dec4-yl)carbamate (15-2, 150 mg, yield 89.8%).

LCMS: m/z 609.0 [M+H] ⁺ .

Step 3: To tert-butyl (S)-(8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazepine Spiro[4.5]dec4-yl)carbamate (15-2,130mg, 0.21mmol) in dioxane (120mL) was added with 2-amino-3-chloropyridine-4-sodium thiosulfate ( C1, 115mg, 0.64mmol), cuprous iodide (40mg, 0.21mmol) and o-phenanthroline (20mg, 0.11mmol), replaced with argon three times, heated to 110°C, and reacted overnight. The reaction solution was concentrated, and column chromatography was carried out to obtain a pale yellow solid tert-butyl (S)-(8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroindole Indole [1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (15-3, 120mg, yield 87.6%).

LCMS: m/z 641.1 [M+H] ⁺ .

Step 4: To tert-butyl (S)-(8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroindole[1,2-c]pyrimidine -5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (15-3, 20mg, 0.03mmol) in dichloromethane (4mL) Trifluoroacetic acid (2mL) was added and reacted for 2 hours at room temperature. The reaction solution was concentrated, diluted with dichloromethane (50mL), washed with saturated sodium bicarbonate, separated, the organic phase was washed with brine, dried, filtered, and concentrated to Dry, purified by preparative chromatography to obtain a white solid (S)-8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroimidazo[1,2-c] Pyrimidine-5-yl)-2-phenyl-2,8-diazaspiro[4.5]decane 4-amine formate (15,5 mg, yield 28.0%).

LCMS: m/z 541.1[M+H] ⁺ ;

¹ H NMR (400MHz, MeOD-d ₄ ) δ8.55 (brs, 1H), 7.84 (s, 1H), 7.54-7.51 (m, 2H), 7.23-7.20 (m, 2H), 6.71-6.65 (m ,3H),5.91(d,J=5.2Hz,1H),4.59-4.57(m,1H),4.17-4.09(m,2H),3.75-3.25(m,6H),2.06-1.97(m,2H) ), 1.88-1.77 (m, 2H).

Example 16: Synthesis of Compound 16.

(S)-8-(7-amino-8-((2-amino-3-chloropyridinyl-4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2- Phenyl-2,8-diazaspiro[4.5]decyl 4-amine

Step 1: To 7-chloro-8-iodo-5-(methylsulfinyl)imidazo[1,2-c]pyrimidine (B5, 222mg, 0.65mmol) and (S)-2-phenyl-2 Add Cs ₂ CO ₃ (2.1g, 6.45mmol) to the suspension of ,8-diazaspiro[4.5]decane 4-amine (3-2, 220mg, 0.72mmol) in acetonitrile (30ml) and raise the temperature to 30°C , Reaction for 1 hour. The reaction solution was diluted with water (50ml), extracted with ethyl acetate (2×30ml), the organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain a white solid (S)-8-(7- Chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec4-amine (16-1, 140mg, yield 42.4%).

LCMS: m/z 509.0 [M+H] ⁺ .

Step 2: To (S)-8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5] Triethylamine (60 mg, 0.56 mmol) and (Boc) ₂ O (120 mg, 0.56 mmol) were added to the solution of decyl 4-amine (16-1, 140 mg, 0.28 mmol) in dichloromethane (20 mL), and reacted at room temperature overnight. The reaction solution was concentrated and purified by column chromatography to obtain pale yellow solid tert-butyl (S)-(8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2- Phenyl-2,8-diazaspiro[4.5]dec4-yl)carbamate (16-2, 150 mg, yield 89.8%).

LCMS: m/z 609.0 [M+H] ⁺ .

Step 3: To tert-butyl (S)-(8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazepine Spiro[4.5]dec4-yl)carbamate (16-2, 130mg, 0.21mmol) in dioxane (120mL) was added with 2-amino-3-chloropyridine-4-sodium thiosulfate ( C1, 115mg, 0.64mmol), cuprous iodide (40mg, 0.21mmol) and o-phenanthroline (20mg, 0.11mmol), replaced with argon three times, heated to 110°C, and reacted overnight. The reaction solution was concentrated, and column chromatography was performed to obtain a pale yellow solid tert-butyl (S)-(8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroindole Indole[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (16-3, 120mg, yield 87.6%).

LCMS: m/z 641.1 [M+H] ⁺ .

Step 4: Under the protection of argon, add tert-butyl(S)-(8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroindole [1, 2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (16-3, 30mg, 0.05mmol) in DMF Sodium azide (12mg, 0.19mmol) and potassium carbonate (26mg, 0.19mmol) were added to the mixture, replaced with argon three times, heated to 100°C, and reacted overnight. The reaction solution was diluted with ethyl acetate (50mL), followed by water ( 3×50mL) and saturated brine (2×50mL) washed, dried, filtered, and concentrated to obtain a pale yellow solid tert-butyl (S)-(8-(7-amino-8-((2-amino-3-chloropyridine) -4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (16-4, 20mg).

LCMS: m/z 622.2 [M+H] ⁺ .

Step 5: To tert-butyl (S)-(8-(7-amino-8-((2-amino-3-chloropyridin-4-yl)thio)imidazo[1,2-c]pyrimidine- 5-yl)-2-phenyl-2,8-diazaspiro[4.5]dec-4-yl)carbamate (16-4, 30mg, 30%, 0.014mmol) in dichloromethane (10mL ) Trifluoroacetic acid (1 mL, 1.35 mmol) was added to the solution and reacted at room temperature for 1 h. The reaction solution was concentrated, diluted with dichloromethane (50mL), washed with saturated sodium bicarbonate and saturated brine successively, dried, filtered, concentrated, and purified by Prep-HPLC to obtain a white solid (S)-8-(7-amino-8) -((2-Amino-3-chloropyridinyl-4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2-phenyl-2,8-diazaspiro[ 4.5] Trifluoroacetate of decyl 4-amine (16, 2.5 mg, yield 30.0%).

LCMS: m/z 522.2[M+H] ⁺ ;

¹ H NMR(400MHz,DMSO-d ₆ +D ₂ O): δ7.72(d,J=2.8Hz,1H), 7.68(d,J=5.6Hz,1H), 7.65(d,J=2.4Hz ,1H),7.23(t,J=7.6Hz,2H),6.69(t,J=7.6Hz,1H),6.63(d,J=7.6Hz,2H),6.19(d,J=5.6Hz,1H ),3.92-3.86(m,2H),3.82-3.74(m,2H),3.47(s,3H),3.45-3.38(m,2H),1.93-1.88(m,2H),1.82-1.77(m ,1H),1.74-1.69(m,1H).

Example 17: Synthesis of Compound 17.

(S)-4-(4-Amino-8-((2-amino-3-chloropyridin-4-yl)sulfanyl)imidazo[1,2-c]pyrimidin-5-yl)-2,8 -Diazaspiro[4.5]-2-yl)benzonitrile

Step 1: To tert-butyl(S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylate ( A1, 100mg, 0.28mmol) in 1,4-dioxane (20mL) solution was sequentially added 4-iodobenzonitrile (128mg, 0.56mmol), Cs ₂ CO ₃ (272mg, 0.42mmol) and Xantphos (32mg , 0.03mmol). Pd ₂ (dba) ₃ (30 mg, 0.01 mmol) was added after 3 times of vacuum nitrogen replacement. After the addition, the vacuum nitrogen is replaced 3 times. Heat to 110°C for 12h. The reaction solution was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to dry to obtain a brown oil, which was purified by silica gel column chromatography to obtain a brown solid tert-butyl (S)-4-(((R)-tert-butylsulfinyl)amino group )-2-(4-cyanophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (17-1, 103 mg, yield: 80.4%). LCMS: m/z 461.6[M+H] ⁺ ; ¹ H NMR(400MHz, CDCl ₃ ), δppm 7.50(d,J=2.8Hz,2H), 7.47(d,J=2.4Hz,2H), 4.00～ 3.97(m,2H),3.79～3.71(m,4H), 3.50(d,J＝9.2Hz,1H), 3.39～3.35(m,2H),3.33(d,J＝5.6Hz,2H), 3.26 (d, J=10.0 Hz, 2H), 1.46 (s, 9H), 1.24 (s, 9H).

Step 2: Add tert-butyl(S)-4-(((R)-tert-butylsulfinyl)amino)-2-(4-cyanophenyl)-2,8-diazepine at 0℃ Spiro[4.5]decane-8-t-butyl carboxylate (17-1, 103mg, 0.22mmol) in dichloromethane (5mL) was added dropwise HCl/1,4-dioxane (4.0M, 0.6mL ). Raise to room temperature and react for 1 hour. The reaction solution was concentrated and dried under reduced pressure to obtain a brown-yellow solid (S)-4-(4-amino-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile hydrochloride (17-2, crude product) , 98mg). LCMS: m/z 257.5 [M+H] ⁺ .

Step 3: To (S)-4-(4-amino-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile (17-2, 98mg, 0.22mmol) in isopropanol ( (12mL) DIPEA (1143mg, 8.85mmol) and 5-chloro-8-iodoimidazo[1,2-c]pyrimidine (138mg, 0.49mmol) were added to the solution. The temperature was raised to 70°C for 4 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=50/1) to obtain a brown-yellow solid (S)-4-(4-amino-8-(8-iodoimidazo[1,2) -c]pyrimidin-5-yl)-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile (17-3, 98 mg, two-step yield: 87.8%). LCMS: m/z 500.4[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d6), δppm 8.05 (s, 1H), 7.87 (s, 1H), 7.66 (s, 1H), 7.56 (d, J = 8.8Hz, 2H), 6.66 (d, J = 8.8Hz, 2H), 3.70 ~ 3.68 (m, 3H), 3.55 (d, J = 10.4 Hz, 1H), 3.48 ~ 3.45 (m, 2H), 3.38(s,1H), 3.24～3.22(m,4H), 1.96～1.86(m,2H), 1.62～1.56(m,2H).

Step 4: To (S)-4-(4-amino-8-(8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2,8-diazaspiro[4.5]dec- 2-yl)benzonitrile (17-3, 95mg, 0.19mmol) in 1,4-dioxane (15mL) solution was added DIPEA (123mg, 0.28mmol), 2-amino-3-chloropyridine- 4-thiol sodium salt (52mg, 0.28mmol) and Xantphos (33mg, 0.057mmol). Pd ₂ (dba) ₃ (27 mg, 0.029 mmol) was added after 3 times of vacuum nitrogen replacement. Vacuum nitrogen replacement 3 times again. The temperature was raised to 100°C and reacted for 4 hours. The reaction solution was diluted with ethyl acetate (20mL), filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by Prep-HPLC to obtain a white solid ((S)-4-(4-amino-8-(8-(((2-amino) -3-Chloropyridinyl-4-yl)thio)imidazolyl[1,2-c]pyrimidin-5-yl)-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile Formate (compound 17, 22mg, yield: 20.0%). LCMS: m/z 532.6[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d6), δ ppm 8.19 (s, 1H), 8.03 ( s, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.58 ~ 7.54 (m, 4H), 6.65 (d, J = 8.8 Hz, 2H), 6.33 (s, 2H), 5.80 (d, J ＝5.2Hz,1H),3.90～3.87(m,2H), 3.65～3.55(m,4H), 3.34～3.32(m,2H), 3.30(s,1H), 3.16～3.08(m,2H), 1.96～1.88(m,2H), 1.59～1.56(m,2H).

Example 18: Synthesis of Compound 18.

(S)-8-(8-((2-Amino-3-chloropyridin-4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2-(4-fluorobenzene Yl)-2,8-diazaspiro[4.5]decane 4-amine

Step 1: To tert-butyl(S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylate ( A1, 75mg, 0.19mmol) in 1,4-dioxane (15mL) solution was sequentially added Cs ₂ CO ₃ (191mg, 0.59mmol), 1-fluoro-4-iodobenzene (87mg, 0.39mmol) and Xantphos (24mg, 0.041mmol). Pd ₂ (dba) ₃ (19 mg, 0.021 mmol) was added after 3 times of vacuum nitrogen replacement. Vacuum nitrogen replacement 3 times again. The temperature was raised to 110°C and reacted for 15 hours. The reaction solution was diluted with ethyl acetate (20mL), filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=80/1) to obtain a brown solid tert-butyl (S)-4-( ((R)-tert-Butylsulfinyl)amino)-2-(4-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate tert-butyl ester (18-1, 92mg, yield: 97.2%). LCMS: m/z 454 [M+H] ⁺ .

Step 2: To tert-butyl (S)-4-(((R)-tert-butylsulfinyl)amino)-2-(4-fluorophenyl)-2,8-diazaspiro[4.5] To a solution of tert-butyl decane-8-carboxylate (18-1, 92 mg, 0.20 mmol) in dichloromethane (5 mL) was added dropwise HCl/1,4-dioxane (4.0M, 0.6 mL). React at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure to dryness to obtain a brown solid (S)-2-(4-fluorophenyl)-2,8-diazaspiro[4.5]dec-4-amine hydrochloride (18-2, crude product) , 73mg). LCMS: m/z 250 [M+H] ⁺ .

Step 3: To (S)-2-(4-fluorophenyl)-2,8-diazaspiro[4.5]dec-4-amine (18-2, 73mg, 0.20mmol) in isopropanol (12mL ) DIPEA (454mg, 3.51mmol) and 5-chloro-8-iodoimidazo[1,2-c]pyrimidine (65mg, 0.23mmol) were added to the solution. The temperature was raised to 70°C and reacted for 4 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by Prep-TLC (dichloromethane/methanol=15/1) to obtain a brown-yellow solid (S)-2-(4-fluorophenyl)-8-(8-iodoimidazo[1 ,2-c]pyrimidin-5-yl)-2,8-diazaspiro[4.5]decane 4-amine (18-3, 60 mg, two-step yield: 60.1%). LCMS: m/z 493 [M+H] ⁺ .

Step 4: To (S)-2-(4-fluorophenyl)-8-(8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2,8-diazaspiro[4.5 ] Decane 4-amine (18-3, 30mg, 0.061mmol) in 1,4-dioxane (15mL) solution was added DIPEA (39mg, 0.30mmol), 2-amino-3-chloropyridine-4- Sodium mercaptan salt (18mg, 0.099mmol) and Xantphos (12mg, 0.020mmol). Pd ₂ (dba) ₃ (9 mg, 0.010 mmol) was added after 3 times of vacuum nitrogen replacement. Vacuum nitrogen replacement 3 times again. The temperature was raised to 100°C and reacted for 4 hours. The reaction solution was diluted with ethyl acetate (20 mL), filtered, and the filtrate was concentrated under reduced pressure to dryness, and purified by Prep-HPLC to obtain a white solid (S)-8-(8-(((2-amino-3-chloropyridin-4-yl) Thio) imidazo[1,2-c]pyrimidin-5-yl)-2-(4-fluorophenyl)-2,8-diazaspiro[4.5]dec4-amine (Compound 18, 2.19mg , Yield: 6.84%). LCMS: m/z 525[M+H] ⁺ ; ¹ H NMR (400MHz, MeOD-d4): δ ppm 8.05 (s, 1H), 7.84 (d, J = 1.6 Hz, 1H ), 7.56 (d, J = 1.6 Hz, 1H), 7.50 (d, J = 5.6 Hz, 1H), 6.95 ~ 6.90 (m, 2H), 6.57 ~ 6.53 (m, 2H), 5.89 (d, J = 5.6Hz, 1H), 4.05～3.99(m,2H), 3.66～3.57(m,2H), 3.43～3.32(m,4H), 3.14～3.11(m,1H), 2.07～1.99(m,2H) , 1.76 ~ 1.67 (m, 2H); ¹⁹ F NMR (376 MHz, MeOD-d4): δ ppm-132.72.

Example 19: Synthesis of Compound 19.

(S)-8-(8-((2-Amino-3-chloropyridin-4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2-(2-chloropyridine -4-yl)-2,8-diazaspiro[4.5]decyl 4-amine

Step 1: To tert-butyl(S)-4-(((R)-tert-butylsulfinyl)amino)-2,8-diazaspiro[4.5]decane-8-carboxylate ( A1, 70mg, 0.19mmol) in 1,4-dioxane (15mL) solution was sequentially added Cs ₂ CO ₃ (191mg, 0.59mmol), 2-chloro-4-iodopyridine (94mg, 0.39mmol) and Xantphos (24mg, 0.041mmol). Pd ₂ (dba) ₃ (19 mg, 0.021 mmol) was added after 3 times of vacuum nitrogen replacement. Vacuum nitrogen replacement 3 times again. The temperature was raised to 110°C and reacted for 15 hours. The reaction solution was diluted with ethyl acetate (20 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=80/1) to obtain a brown solid tert-butyl (S)-4-(( (R)-tert-Butylsulfinyl)amino)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (19- 1,75mg, yield: 81.8%). LCMS: m/z 471.5[M+H] ⁺ ; ¹ H NMR (400MHz, CDCl ₃ ): δppm 8.00(d,J=6.0Hz,1H), 8.40(d,J=6.4Hz,1H), 6.33～ 6.31(m,1H), 4.00～3.97(m,2H), 3.80～3.74(m,1H), 3.72～3.68(m,1H), 3.46～3.45(m,1H), 3.38～3.33(m,2H) ), 3.24 (d, J = 10.0 Hz, 1H), 2.96 to 2.89 (m, 2H), 1.81 to 1.67 (m, 3H), 1.47 (s, 9H), 1.22 (s, 9H).

Step 2: To tert-butyl (S)-4-(((R)-tert-butylsulfinyl)amino)-2-(2-chloropyridin-4-yl)-2,8-diazepine [4.5] To a solution of tert-butyl decane-8-carboxylate (19-1, 75 mg, 0.16 mmol) in dichloromethane (5 mL) was added HCl/1,4-dioxane (4.0M, 0.6 mL). React at room temperature for 1.5 hours. The reaction solution was concentrated and dried under reduced pressure to obtain a brown-yellow solid (S)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-amine hydrochloride (19-2 , Crude product, 77mg).

LCMS: m/z 267.4[M+H] ⁺

Step 3: To (S)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-amine (19-2, 75mg, 0.16mmol) in isopropyl DIPEA (420mg, 3.25mmol) and 5-chloro-8-iodoimidazo[1,2-c]pyrimidine (60mg, 0.21mmol) were added to the alcohol (12mL) solution. The temperature was raised to 70°C and reacted for 4 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by Prep-TLC (dichloromethane/methanol=15/1) to obtain a brown solid (S)-2-(2-chloropyridin-4-yl)-8-(8-iodine) Imidazo[1,2-c]pyrimidin-5-yl)-2,8-diazaspiro[4.5]dec-4-amine (19-3, 46mg, two-step yield: 56.7%). LCMS: m/z 510.3[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ): δppm 8.05(s, 1H), 7.91(d, J=5.6Hz, 1H), 7.87(s, 1H ),7.66(s,1H),6.53(s,2H),3.70～3.38(m,8H),3.27～3.18(m,3H),1.92～1.88(m,2H),1.61～1.54(m,2H) ).

Step 4: To (S)-2-(2-chloropyridin-4-yl)-8-(8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2,8-diazepine Spiro[4.5]dec-4-amine (46mg, 0.09mmol) in 1,4-dioxane (15mL) solution was added DIPEA (59mg, 0.46mmol), 2-amino-3-chloropyridine-4- Sodium mercaptan salt (26mg, 0.14mmol) and Xantphos (17mg, 0.029mmol). Pd ₂ (dba) ₃ (13 mg, 0.015 mmol) was added after 3 times of vacuum nitrogen replacement. Vacuum nitrogen replacement 3 times again. The temperature was raised to 100°C and reacted for 4 hours. The reaction solution was diluted with ethyl acetate (20 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure, and purified by Prep-HPLC to obtain a white solid (S)-8-(8-(((2-amino-3-chloropyridine-4- (Yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]deca-4-carboxamide Acid salt (compound 19, 19.83 mg, yield: 37.4%). LCMS: m/z 542.6[M+H] ⁺ ; ¹ H NMR (400MHz, MeOD-d ₄ ): δppm8.42(s, 1H), 8.06(s,1H),7.92(d,J=6.4Hz,1H),7.84(d,J=5.6Hz,1H),7.57(d,J=1.6Hz,1H),7.50(d,J=5.2 Hz,1H),6.63(s,1H),6.59(d,J=5.6Hz,1H),5.89～5.86(m,1H),4.05～3.99(m,2H),3.85～3.81(m,1H) , 3.64～3.53(m,3H), 3.48～3.36(m,3H), 2.10～2.00(m,2H), 1.81～1.74(m,2H).

Example 20: Synthesis of Compound 20.

(S)-8-(7-amino-8-((2-amino-3-chloropyridinyl-4-yl)thio)imidazo[1,2-c]pyrimidin-5-yl)-2- (2-Chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec4-amine

Step 1: Add 6-amino-2-chloro-5-iodopyrimidine-4-carboxylic acid methyl ester (20-1, 1.0 g, 3.19 mmol) to chloroacetaldehyde (40 wt.% aqueous solution, 12 mL). Heat to 80°C to react for 2 hours. After the reaction solution was cooled to room temperature, it was extracted with ethyl acetate (3×80 mL). The extracts were combined, washed with water (3×80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. Ethyl ester = 3/1) was purified to give a pale yellow solid 5-chloro-8-iodoimidazo[1,2-c]pyrimidine-7-carboxylic acid methyl ester (20-2,537 mg, yield: 50%). ¹ H NMR (400MHz, CDCl ₃ ): δ ppm 7.98 (d, J = 1.6 Hz, 1H), 7.93 (d, J = 1.6 Hz, 1H), 4.04 (s, 3H).

Step 2: To (R)-N-((S)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-yl)-2-methyl Propane-2-sulfinamide (130mg, 0.35mmol) in isopropanol (20mL) was added DIPEA (135mg, 1.05mmol) and 5-chloro-8-iodoimidazo[1,2-c]pyrimidine- Methyl 7-carboxylate (20-2, 132 mg, 0.39 mmol). After purging with vacuum nitrogen 3 times, the temperature was raised to 70°C and reacted for 3 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=25/1) to obtain a pale yellow solid methyl 5-((S)-4-(((R)-tert-butylsulfinyl) )Amino)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine-7 -Carboxylate (20-3,200 mg, yield: 84.7%). LCMS: m/z 672.1 [M+H] ⁺ .

Step 3: To methyl 5-((S)-4-(((R)-tert-butylsulfinyl)amino)-2-(2-chloropyridin-4-yl)-2,8-diazepine Heterosspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine-7-carboxylate (20-3,200mg, 0.30mmol) of 1,4-dioxane (15mL) DIPEA (114mg, 0.89mmol) and 2-amino-3-chloropyridine-4-thiol sodium salt (108mg, 0.60mmol) were added to the solution in sequence. After 3 times of vacuum nitrogen replacement, Xantphos (34 mg, 0.06 mmol) and Pd ₂ (dba) ₃ (55 mg, 0.06 mmol) were added. Vacuum nitrogen replacement 3 times again. The temperature was raised to 110°C to react for 4 hours. The reaction solution was diluted with ethyl acetate (50 mL), filtered, the filtrate was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=15/1) to obtain a pale yellow solid methyl 8-((2-amino- 3-chloropyridin-4-yl)sulfanyl)-5-((S)-4-(((R)-tert-butylsulfinyl)amino)-2-(2-chloropyridinyl-4-yl )-2,8-diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine-7-carboxylate (20-4,160mg, yield: 76.3% ). LCMS: m/z 704.1 [M+H] ⁺ .

Step 4: To methyl 8-((2-amino-3-chloropyridin-4-yl)sulfanyl)-5-((S)-4-(((R)-tert-butylsulfinyl)amino )-2-(2-Chloropyridyl-4-yl)-2,8-diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine-7- To a solution of carboxylate (20-4, 160mg, 0.23mmol) in dichloromethane (20mL) was added di-tert-butyl dicarbonate (250mg, 1.14mmol), triethylamine (70mg, 0.68mmol) and DMAP (56mg, 0.46mmol). React overnight at room temperature. The reaction solution was concentrated under reduced pressure to dryness, and purified by silica gel column chromatography (dichloromethane/methanol=30/1) to obtain a brown solid methyl 8-((2-tert-butoxycarbonylamino-3-chloropyridine-4- Yl)thio)-5-((S)-4-(((R)-di-tert-butylsulfinyl)amino)-2-(2-chloropyridyl-4-yl)-2,8- Diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine-7-carboxylate (20-5,100 mg, yield: 48.7%).

LCMS: m/z 905.7 [M+H] ⁺ .

Step 5: To methyl 8-((2-di-tert-butoxycarbonylamino-3-chloropyridin-4-yl)sulfanyl)-5-((S)-4-(((R)-tert-butyl Sulfinyl)amino)-2-(2-chloropyridyl-4-yl)-2,8-diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c ] Pyrimidine-7-carboxylate (20-5, 100 mg, 0.11 mmol) and sodium hydroxide (18 mg, 0.45 mmol) in methanol (15 mL) was added with water (3 mL). The temperature was raised to 50°C to react for 1 hour, neutralized with 2M dilute hydrochloric acid, and concentrated under reduced pressure. Purified by silica gel column chromatography (dichloromethane/methanol=8/1) to obtain a yellow solid 8-((2-di-tert-butoxycarbonylamino-3-chloropyridin-4-yl)sulfanyl)-5-( (S)-4-(((R)-tert-butylsulfinyl)amino)-2-(2-chloropyridyl-4-yl)-2,8-diazaspiro[4.5]dec-8 -Yl)-8-iodoimidazo[1,2-c]pyrimidine-7-carboxylic acid (20-6, 90 mg, yield: 91.8%). LCMS: m/z 890.3 [M+H] ⁺ .

Step 6: To 8-((2-di-tert-butoxycarbonylamino-3-chloropyridin-4-yl)sulfanyl)-5-((S)-4-(((R)-tert-butylsulfinyl) Acyl)amino)-2-(2-chloropyridyl-4-yl)-2,8-diazaspiro[4.5]dec-8-yl)-8-iodoimidazo[1,2-c]pyrimidine To a solution of -7-carboxylic acid (20-6, 90 mg, 0.10 mmol) in tert-butanol (20 mL) was added triethylamine (102 mg, 1.00 mmol) and DPPA (83 mg, 0.30 mmol). After purging with vacuum nitrogen 3 times, the temperature was raised to 100°C and reacted for 3 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=25/1) to obtain a pale yellow solid tert-butyl (8-(((2-di-tert-butoxycarbonylamino-3-chloropyridine- 4-yl)thio)-5-((S)-4-(((R)-tert-butylsulfinyl)amino)-2-(2-chloropyridin-4-yl)-2,8- Diazaspiro[4.5]dec-8-yl)imidazo[1,2-c]pyrimidin-7-yl)carbamate (20-7,70 mg, purity 50.6%, yield: 40.6%). LCMS: m/z 862.7 [M+H] ⁺ .

Step 7: To tert-butyl (8-(((2-di-tert-butoxycarbonylamino-3-chloropyridin-4-yl)sulfanyl)-5-((S)-4-(((R)- Tert-Butylsulfinyl)amino)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-8-yl)imidazo[1,2-c]pyrimidine -7-yl) carbamate (20-6, 70mg, purity 50.6%, 0.04mmol) in dichloromethane (20 mL) was added HCl/dioxane (3M, 4mL). Reaction at room temperature for 1.5 hours The reaction solution was adjusted to pH 8 with sodium bicarbonate solution, extracted with dichloromethane/methanol (10/1) (3x100mL), the organic phase was concentrated and purified by silica gel column chromatography (dichloromethane/methanol=7/1) And Prep-HPLC to obtain white solid (S)-8-(7-amino-8-((2-amino-3-chloropyridin-4-yl)sulfanyl)imidazo[1,2-c]pyrimidine-5 -Yl)-2-(2-chloropyridyl-4-yl)-2,8-diazaspiro[4.5]dec4-amine (Compound 20, 1 mg, yield: 4.5%). LCMS: m/ z 557.1[M+H] ⁺ .

¹ H NMR (400MHz, MeOD-d ₄ ): δppm 7.88 (d, J = 6.0 Hz, 1H), 7.52 (d, J = 5.2 Hz, 1H), 7.48 (d, J = 1.6 Hz, 1H), 7.32 ～7.20(m,1H), 7.19(d,J=1.6Hz,1H), 6.57～6.54(m,2H), 5.98(d,J=5.6Hz,1H), 3.98～3.93(m,2H), 3.73～3.69(m,1H), 3.64～3.61(m,1H), 3.48～3.35(m,2H), 3.23～3.12(m,2H), 2.04～1.98(m,2H), 1.71～1.63(m ,2H).

Example 21: Synthesis of Compound 21.

(S)-8-(8-(((2-Amino-3-chloropyridin-4-yl)thio)-7-chloroimidazo[1,2-c]pyrimidin-5-yl)-2- (2-Chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec4-amine

Step 1: To N-((S)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-yl)-2-methylpropane-2- DIPEA (145mg, 1.12mmol) and 7-chloro-8-iodo-5-(methylsulfinyl) imidazo[1, sulfinamide (83mg, 0.22mmol) in dichloromethane (20mL) solution were added successively 2-c] A solution of pyrimidine (154 mg, 0.45 mmol) in dichloromethane (3 mL). After 3 times of vacuum nitrogen replacement, the reaction was carried out at room temperature for 2 hours. The reaction solution was concentrated to dryness under reduced pressure, and purified by silica gel column chromatography (dichloromethane/methanol=30/1) to obtain light yellow solid N-((S)-8-(7-chloro-8-iodoimidazo[1,2 -c]pyrimidin-5-yl)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-yl)-2-methylpropane-2-ylidene Sulfonamide (40 mg, yield: 27.6%). LCMS: m/z 648.0 [M+H] ⁺ .

Step 2: To N-((S)-8-(7-chloro-8-iodoimidazo[1,2-c]pyrimidin-5-yl)-2-(2-chloropyridin-4-yl)- 2,8-Diazaspiro[4.5]dec-4-yl)-2-methylpropane-2-sulfenamide (40mg, 0.06mmol) in 1,4-dioxane (15mL) solution DIPEA (0.2 mL, 1.98 mmol), 2-amino-3-chloropyridine-4-thiol sodium salt (18 mg, 0.10 mmol) was added. After 3 times of vacuum nitrogen replacement, Xantphos (8 mg, 0.01 mmol) and Pd ₂ (dba) ₃ (16 mg, 0.02 mmol) were added. Vacuum nitrogen replacement 3 times again. The temperature was raised to 110°C for 16 hours. The reaction solution was diluted with ethyl acetate (50 mL), filtered, and the filtrate was concentrated under reduced pressure to dryness, and purified by silica gel column chromatography (dichloromethane/methanol=10/1) to obtain a pale yellow solid N-((S)-8-( 8-(((2-Amino-3-chloropyridin-4-yl)thio)-7-chloroimidazole[1,2-c]pyrimidin-5-yl)-2-(2-chloropyridine-4- Yl)-2,8-diazaspiro[4.5]dec-4-yl)-2-methylpropane-2-sulfinamide (22mg, yield: 52.4%). LCMS: m/z 682.1[M +H] ⁺ .

Step 3: To N-((S)-8-(8-(((2-amino-3-chloropyridin-4-yl)thio)-7-chloroimidazole[1,2-c]pyrimidine-5 -Yl)-2-(2-chloropyridin-4-yl)-2,8-diazaspiro[4.5]dec-4-yl)-2-methylpropane-2-sulfenamide (22mg, 0.03 HCl/dioxane (1mL) was added to the dichloromethane (4mL) solution of 1 mmol). Reacted at room temperature for 1 hour. The pH of the reaction solution was adjusted to 8 with sodium bicarbonate solution, and then dichloromethane/methanol (10/1) After extraction (3×50mL), the organic phase was concentrated and purified by silica gel column chromatography (dichloromethane/methanol=7/1) and Prep-HPLC to obtain a white solid (S)-8-(8-(((2-amino -3-Chloropyridin-4-yl)thio)-7-chloroimidazo[1,2-c]pyrimidin-5-yl)-2-(2-chloropyridin-4-yl)-2,8 -Diazaspiro[4.5]dec-4-amine to give the formate (2.0mg, yield: 9.9%). LCMS: m/z 578.0[M+H] ⁺ . ¹ H NMR (400MHz, MeOD-d ₄ ): δppm 8.43 (br, 1H), 7.92 (d, J = 6.0 Hz, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.54 (d, J = 1.6 Hz, 1H), 7.53 (d ,J=5.6Hz,1H),6.62(s,1H),8.59～8.58(m,1H),5.92(d,J＝6.0Hz,1H),4.15～4.09(m,2H),3.83～3.79( m,1H), 3.65～3.46(m,6H), 2.09～1.98(m,2H), 1.80～1.73(m,2H).

Example 22: Synthesis of Compound 22-29.

Using the same method as in Example 21, the following compounds 22-29 were prepared with corresponding reactants:

The biological function of the compound disclosed in the present invention has been proved in the test of enzyme activity and cell level. For example, in the SHP2 enzyme activity inhibition test, the compound disclosed in the present invention can achieve strong inhibitory activity (IC50 up to <10 nM). At the cellular level, the compound disclosed in the present invention can not only inhibit the phosphorylation level of downstream pathway kinase ERK, but also exhibit a good activity of inhibiting the proliferation of cancer cells. Compared with SHP099 (6-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)pyrazine-2-amine), the compound of the present invention is no matter in the enzyme Both on the scientific level and on the cellular level have demonstrated superior activity.

Test case 1: SHP2 enzyme activity test

The compound powder is dissolved in DMSO to make a mother liquor. In the experiment, the compound stock solution was diluted with DMSO in 3-fold gradient, and the same compound was set to 10 different test concentrations. Take 1 μL of the compound at each concentration point into the well of the detection plate (Corning, Costar 3915), and set 2 parallel replicates for each concentration point. Using 6,8-difluoro-4-methyl-7-hydroxycoumarin phosphate (DiFMUP) as a substrate, SHP2 E72A catalyzes its hydrolysis to produce 6,8-difluoro-4-methyl-7-hydroxy Bean (DiFMU), through the PE Enspire multi-function reader, use 358nm as the excitation wavelength to detect the fluorescence value at 455nm to determine the enzyme activity of SHP2.

The composition of the SHP2 buffer used in the reaction is 60mmol/L Hepes, PH7.2, 75mmol/L NaCl, 75mmol/L KCl, 1mmol/L EDTA, 5mmol/L DTT. The screening system consists of: SHP2 buffer, enzyme SHP2 E76A protein, substrate DiFMUP and test compound.

IC ₅₀ test method:

In a 96-well screening plate, 50ng SHP2 E76A protein was reacted with the test compound in SHP2 buffer for 20 minutes, and then incubated with 10uM DiFMUP for 20 minutes at room temperature. The PE Enspire multi-function reader was used to excite light at 358nm and read the light at 455nm. strength. The fluorescence value measured by the compound treatment group is compared with the value of the DMSO control well to calculate the inhibition rate of the sample on the enzyme activity. The IC 50 value of the compound was calculated by using the Prism software of Graphpad Company to fit the inhibition rate to the concentration of the inhibitor nonlinearly. The curve of enzyme activity varying with compound concentration was fitted by the equation Y=Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope)). The IC ₅₀ value of each compound was obtained. Table 1 below shows the IC ₅₀ values of some compounds of the present invention. Among them, the letter A represents that the IC _{50 is} less than 50 nM; the letter B represents that the IC ₅₀ is 50 nM to 500 nM.

Table 1

化合物编号Compound number	酶活性(nM)Enzyme activity (nM)	化合物编号Compound number	酶活性(nM)Enzyme activity (nM)
SHP099SHP099	263263	化合物15Compound 15	AA
化合物1Compound 1	AA	化合物16Compound 16	AA
化合物2Compound 2	AA	化合物17Compound 17	AA
化合物3Compound 3	AA	化合物18Compound 18	AA
化合物4Compound 4	AA	化合物19Compound 19	AA
化合物5Compound 5	AA	化合物20Compound 20	AA
化合物6Compound 6	AA	化合物21Compound 21	AA
化合物7Compound 7	BB	化合物22Compound 22	AA
化合物8Compound 8	AA	化合物23Compound 23	AA
化合物9Compound 9	AA	化合物24Compound 24	AA
化合物10Compound 10	AA	化合物25Compound 25	AA
化合物11Compound 11	BB	化合物26Compound 26	AA
化合物12Compound 12	BB	化合物27Compound 27	AA
化合物13Compound 13	AA	化合物28Compound 28	AA
化合物14Compound 14	AA	化合物29Compound 29	AA

Test Example 2: MV4-11 cell proliferation inhibition experiment

by

The luminescence cell viability detection kit quantitatively measures intracellular ATP to detect the number of viable cells in culture.

The first step: Seed MV4-11 cells in a 96-well plate, seed the cells into a 96-well plate at a density of 2500 cells per well, with a volume of 100 μL per well. Place it in a 37°C, 5% carbon dioxide incubator overnight.

Step 2: Compound treatment of cells. The test compound was diluted 3 times, and a total of 8 concentration gradients were set; a certain volume of DMSO or test compound was added to each well, and each concentration was set to 2 replicates, and the final concentration of DMSO was controlled at 0.5%. Place in a 37°C 5% carbon dioxide incubator for 72h.

Step 3: Use

Luminescent Cell Viability Assay Kit (Promega, G7570) detects the cell viability of the control and treatment groups. Add 50ul CellTiter-Glo to each well, mix well, and incubate at room temperature for 10 minutes. Use EnSpire (Perkin Elmer) to read the signal. The inhibition percentage (%) is calculated by the following formula:

Inhibition percentage (%)=(1-compound treatment group signal value/DMSO treatment group signal value)*100.

The results are shown in Table 2. Among them, the letter A represents that the IC _{50 is} less than 100 nM; the letter B represents that the IC ₅₀ is 100 nM to 500 nM.

Table 2: Compounds inhibiting the proliferation of MV411

Test Example 3: KYSE520 cell proliferation inhibition experiment

CellTiter-Glo ^R luminescence cell viability detection kit is used to quantitatively determine intracellular ATP to detect the number of viable cells in culture.

The first step: Seed KYSE520 cells in a 96-well plate, seed the cells in a 96-well plate at a density of 1500 cells per well, with a volume of 100 μL per well. Place it in a 37°C, 5% carbon dioxide incubator overnight.

Step 2: Compound treatment of cells. The test compound was diluted 3 times, and a total of 8 concentration gradients were set; a certain volume of DMSO or test compound was added to each well, and each concentration was set to 2 replicates, and the final concentration of DMSO was controlled at 0.5%. Place in a 37°C 5% carbon dioxide incubator for 72 hours.

The third step: use CellTiter-Glo ^R Luminescent Cell Viability Assay Kit (Promega, G7570) to detect the cell viability of the control group and the treatment group. Add 50ul CellTiter-Glo to each well, mix well, and incubate at room temperature for 10 minutes. Use EnSpire (Perkin Elmer) to read the signal. The percentage inhibition (%) is calculated by the following formula: percentage inhibition (%)=(1-compound treatment group signal value/DMSO treatment group signal value)*100. The results are shown in Table 3 below.

Among them, the letter A represents that the IC _{50 is} less than 1000 nM; the letter B represents that the IC ₅₀ is between 1000 nM and 2000 nM;

table 3

化合物编号Compound number	KYSE520细胞增殖(nM)KYSE520 cell proliferation (nM)
SHP099SHP099	68696869
化合物7Compound 7	BB

化合物8Compound 8	AA
化合物10Compound 10	AA
化合物16Compound 16	AA
化合物17Compound 17	BB
化合物18Compound 18	AA
化合物19Compound 19	AA
化合物20Compound 20	AA
化合物21Compound 21	AA

Test Example 4: Compound Pharmacokinetic Experiment

The following methods were used to determine the pharmacokinetic parameters of the compounds of this application. The healthy male adult rats/mice used in the study were given a single intragastric administration of 5-100 mg/Kg for each group of animals. Fasting is from 10 hours before administration to 4 hours after administration. Blood was collected at different time points after administration, and the plasma content of the compound was determined (LC-MS/MS). Use professional software to analyze (winnonlin) plasma concentration-time relationship and calculate the pharmacokinetic parameters of the compound. The results show that the compound of the present invention has excellent pharmacokinetic properties.

All documents mentioned in the present invention are cited as references in this application, as if each document is individually cited as a reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A compound represented by Formula I, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof:

among them,

R 1 is selected from H, -halogen, -CN, -OH, -NO 2 , HSO 3 -, unsubstituted or substituted C1-C6 alkylsulfonyl, unsubstituted or substituted C1-C6 alkylcarboxy, unsubstituted Or substituted C1-C6 alkylamino, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, unsubstituted or substituted C1-6 alkoxycarbonyl, unsubstituted or substituted C1-6 alkyl carbonyl group, unsubstituted or substituted C2-C6 alkenyl carbonyl group, unsubstituted or substituted C1-C6 alkoxy-O-C1-C6 alkyl group, unsubstituted or substituted 3-8 membered ring Alkyl group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5-10 membered heteroaryl group; the heterocyclic group or heteroaryl group The group contains 1-4 heteroatoms selected from the group consisting of N, O or S;

R 2 is selected from H, -halogen, amino, unsubstituted or substituted C1-C6 alkylamino, unsubstituted or substituted C1-C6 alkyl;

R 3 is selected from H, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group, substituted or unsubstituted 5-10 membered aryl, substituted or unsubstituted 5 -10 membered heteroaryl; said heterocyclic group or heteroaryl group contains 1-4 heteroatoms selected from the group consisting of N, O or S;

X and Y are each independently selected from N or CR 4 ;

R 4 is selected from H, halogen, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, (substituted or unsubstituted C1-C6 Alkyl)SO-, (substituted or unsubstituted C1-C6 alkyl)SO 2 -;

Or R 4 and adjacent
Together to form a substituted or unsubstituted 5-10 membered aromatic ring, substituted or unsubstituted 5-10 membered heteroaromatic ring, substituted or unsubstituted 5-10 membered heterocyclic group or substituted or unsubstituted 5-10 membered ring Carbocyclyl; the ring contains 0-4 heteroatoms selected from the group consisting of N, O or S; the condition is that X and Y cannot be N at the same time;

Represents a single bond or a double bond;

Any of the above "substituted" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: -D, halogen, -OH, -NO 2 , -NH 2 , -NH (unsubstituted or Halogenated C1-C6 alkyl), -N (unsubstituted or halogenated C1-C6 alkyl) 2 , -CN, unsubstituted or halogenated C1-C8 alkyl, unsubstituted or halogenated C1- C8 alkoxy, unsubstituted or halogenated C1-C8 alkoxy-C1-C8 alkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C3-C8 cycloalkyl- C1-C8 alkyl, unsubstituted or halogenated C1-C6 alkylcarbonyl, unsubstituted or halogenated C1-C6 alkoxycarbonyl, hydroxamic acid group, unsubstituted or halogenated C1-C6 alkyl Mercapto, -S(O) 2 N (unsubstituted or halogenated C1-C6 alkyl) 2 , -S(O) 2 unsubstituted or halogenated C1-C6 alkyl, -N (unsubstituted or halogenated C1-C6 alkyl) S(O) 2 N (unsubstituted or halogenated C1-C6 alkyl) 2 , -S(O)N (unsubstituted or halogenated C1-C6 alkyl) 2 ,- S(O) (unsubstituted or halogenated C1-C6 alkyl), -NH (unsubstituted or halogenated C1-C6 alkyl) S(O)N (unsubstituted or halogenated C1-C6 alkyl) ) 2 , -N (unsubstituted or halogenated C1-C6 alkyl) 2 S(O) (unsubstituted or halogenated C1-C6 alkyl), unsubstituted or halogenated 5-8 membered aryl, Unsubstituted or halogenated 5-8 membered heteroaryl, unsubstituted or halogenated 4-8 membered saturated heterocyclic ring, unsubstituted or halogenated 4-8 membered saturated carbocyclic ring; wherein, the heteroaryl group Or the saturated heterocyclic ring contains 1-4 heteroatoms selected from the group consisting of N, O or S.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof is represented by the following formula II:

among them,

X and Y are each independently selected from N or C;

R 4a and R 4b are each independently selected from none, H, halogen, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, ( Substituted or unsubstituted C1-C6 alkyl) SO-, (substituted or unsubstituted C1-C6 alkyl) SO 2 -;

Other substituents are as described in claim 1.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof is represented by the following formula III:

among them,

X and Y are independently selected from N or C; X and Y cannot be N at the same time;

The ring A is a substituted or unsubstituted 5-10 membered aromatic ring, a substituted or unsubstituted 5-10 membered heteroaromatic ring, a substituted or unsubstituted 5-10 membered heterocyclic group, or a substituted or unsubstituted 5-10 membered carbocyclic group; the heteroaromatic ring or heterocyclic group contains 0-4 heteroatoms selected from the group consisting of N, O or S;

The above-mentioned "substituted" means that one or more hydrogen atoms on ring A are substituted by a substituent selected from the following group: -D, halogen, -OH, -NO 2 , -NH 2 , -NH (unsubstituted or halogenated C1-C6 alkyl), -N (unsubstituted or halogenated C1-C6 alkyl) 2 , -CN, halogenated or unsubstituted C1-C6 alkyl, halogenated or unsubstituted C1-C6 alkoxy Group, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO-, (halogenated or unsubstituted C1-C6 alkyl) SO 2 -.

Other substituents are as described in claim 1.
The compound according to claim 3, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug, or metabolite thereof, wherein the ring A is selected from substituted Or unsubstituted three-membered (hetero) ring, substituted or unsubstituted four-membered (hetero) ring, substituted or unsubstituted five-membered (hetero) ring, substituted or unsubstituted six-membered (hetero) ring, substituted or unsubstituted Substituted five-membered aromatic (hetero) ring, substituted or unsubstituted six-membered aromatic (hetero) ring, substituted or unsubstituted seven-membered aromatic (hetero) ring, substituted or unsubstituted five-membered and six-membered (hetero) ring , Substituted or unsubstituted six-membered and six-membered (hetero) ring; the heterocyclic ring contains 1-4 heteroatoms selected from the group consisting of N, O or S; when the ring A is substituted, the substituent Selected from: halogen, -CN, -OH, -NH 2 , (halogenated or unsubstituted C1-C6 alkyl) 2 N-, halogenated or unsubstituted C1-C6 alkyl, halogenated or unsubstituted C1-C6 alkoxy, C1-C6 alkylamino, (halogenated or unsubstituted C1-C6 alkyl) SO-, (halogenated or unsubstituted C1-C6 alkyl) SO 2 -.
The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, wherein R 1 Selected from H, halogenated or unsubstituted C1-C6 alkylcarbonyl, halogenated or unsubstituted C2-C6 alkenylcarbonyl, (halogenated or unsubstituted C1-C6 alkyl) SO 2 -, substituted or unsubstituted Substituted benzenesulfonyl group, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5 -10 membered heteroaryl group; wherein, when the benzenesulfonyl group, carbocyclic group, heterocyclic group, aryl group and heteroaryl group are substituted, the number of substituents is 1-4, selected from H, NH 2 , Halogen, halogenated or unsubstituted C1-C8 cycloalkyl, unsubstituted or halogenated C1-C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) 2 N-.
The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof, wherein R 3 Selected from H, unsubstituted or substituted 3-8 membered carbocyclic group, unsubstituted or substituted 3-8 membered heterocyclic group, unsubstituted or substituted 5-10 membered aryl group, unsubstituted or substituted 5-10 Membered heteroaryl group; wherein, when the carbocyclic group, heterocyclic group, aryl group and heteroaryl group are substituted, the number of substituents is 1-4, and the substituents are selected from H, NH 2 , halogen, and Substituted or halogenated C1-C6 alkyl, (halogenated or unsubstituted C1-C6 alkyl) 2 N- and halogenated or unsubstituted C1-C6 alkyl -NH-.
The compound of claim 3, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug, or metabolite thereof, wherein in the formula III,

R 1 is selected from 5-10 membered heteroaryl groups substituted with optionally halogen substituents, preferably 5-10 membered nitrogen-containing heteroaryl groups, more preferably pyridyl;

R 2 is selected from H and amino;

R 3 is selected from 5-10 membered heteroaryl groups optionally substituted by 1 or 2 substituents selected from NH 2 and halogen, preferably 5-10 membered nitrogen-containing heteroaryl groups, more preferably pyridyl;

Ring A is:

Among them, X is N and Y is C.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug, or metabolite thereof, characterized in that the compound is selected from the following:
The use of the compound of formula I according to any one of claims 1-8, characterized in that it is used for:

(a) Preparation of drugs for the prevention or treatment of diseases or disorders related to abnormal SHP2 activity;

(b) Preparation of drugs for the prevention or treatment of SHP2-mediated diseases or conditions

(c) Preparation of inhibitor drugs that inhibit SHP2 activity;

(d) Non-therapeutic inhibition of SHP2 activity in vitro;

(e) Non-therapeutic inhibition of tumor cell proliferation in vitro; and/or

(f) Treatment of diseases or conditions related to abnormal SHP2.
The use according to claim 9, wherein the disease is cancer, including but not limited to Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid Leukemia, breast cancer, esophageal cancer, lung cancer, colon cancer, head cancer, neuroblastoma, squamous cell carcinoma of the head and neck, gastric cancer, anaplastic large cell lymphoma or glioblastoma.
A pharmaceutical composition, characterized in that the pharmaceutical composition comprises:

(i) An effective amount of the compound of formula I according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, or a solvate, isotope substitution, polymorph, prodrug or metabolite thereof ;with

(ii) A pharmaceutically acceptable carrier.
A method for inhibiting the activity of SHP2, which is characterized by comprising the steps of: administering an inhibitory effective amount of the compound of formula I according to any one of claims 1 to 8 or an acceptable salt thereof, Or administer an effective amount of the pharmaceutical composition according to claim 11 to a subject to be inhibited.