2-aminopyrimidine compounds and their application and pharmaceutical composition

Abstract

The present invention relates to a 2-aminopyrimidine compound and its application and pharmaceutical composition. The compound can inhibit the activities of CDK4 and CDK6, especially can highly selectively inhibit the activity of CDK4; can treat or prevent transitional proliferative diseases such as cancer mediated by CDK4 and CDK6, especially treat or prevent transitional proliferative diseases such as cancer mediated by CDK4.

Description

2-Aminopyrimidine compounds and their applications and pharmaceutical compositions Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a 2-aminopyrimidine compound and its application and pharmaceutical composition. Background Art

[0002] Protein kinases (PKs) are enzymes that catalyze protein phosphorylation. They are widely present in the human body and play important physiological roles. Protein kinases include mitogen-activated protein kinases, cell cycle-dependent kinases, and tyrosine protein kinases. Cyclin-dependent kinases (CDKs) belong to the serine / threonine kinase family and are involved in physiological processes such as cell proliferation and transcription. CDKs can be divided into two major categories based on their functions: one class, including CDK1, CDK2, CDK4, and CDK6, participates in cell cycle regulation; the other class, including CDK7, CDK8, CDK9, CDK12, and CDK13, participates in transcriptional regulation. Abnormal regulation or mutation of protein kinases can often lead to hyperproliferative diseases, cardiovascular and cerebrovascular diseases, diabetes, inflammatory diseases, and immune system disorders. Therefore, developing compounds that can inhibit CDK activity is of great significance.

[0003] Summary of the Invention

[0004] Based on this, the present invention provides a 2-aminopyrimidine compound capable of inhibiting the activity of CDK4 and CDK6, and its application and pharmaceutical composition.

[0005] The technical solution of the present invention to solve the above technical problems is as follows.

[0006] A 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0007] in:

[0008] R1 is independently selected from halogen, cyano, trihalomethyl, nitro, carboxyl or acyl, and n1 is selected from 1 or 2;

[0009] R2, R4, and R5 are independently selected from H, halogen, C1-C4 alkyl, and C1-C4 alkoxy, n2 is selected from 1 or 2, n4 is selected from an integer of 1 to 4, and n5 is selected from an integer of 1 to 3;

[0010] R3 is selected from H, C1-C4 alkyl, C1-C4 alkoxy, (dialkylamino)methyl, acyl, hydroxy-substituted alkyl;

[0011] X represents O or NR6;

[0012] R6 is selected from alkylsulfonyl;

[0013] Y represents N or CR7;

[0014] R7 is selected from halogen, cyano, trihalomethyl, nitro, carboxyl, and acyl.

[0015] In some embodiments, the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the general formula (I) is the structure of formula (I-1):

[0016] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R1 is selected from halogen and cyano.

[0017] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R1 is selected from Cl and cyano.

[0018] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R3 is selected from C1-C4 alkyl, (dialkylamino)methyl, acyl, or hydroxy-substituted alkyl.

[0019] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R3 is selected from methyl, (dimethylamino)methyl, acetyl, and 2-hydroxyisopropyl.

[0020] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R2 is H.

[0021] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R4 is H.

[0022] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R5 is H.

[0023] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, X represents O or NR6; and R6 is methylsulfonyl.

[0024] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, Y represents N or CR7; and R7 is selected from halogen and cyano.

[0025] In some embodiments, in the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, Y represents N or CR7; and R7 is selected from F and cyano.

[0026] In some embodiments, the 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the general formula (I) is selected from any one of formulas (II-1) to (II-9):

[0027] The present invention provides use of the above-mentioned 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of protein kinase inhibitors, wherein the protein kinase inhibitors include CDK4 inhibitors and CDK6 inhibitors.

[0028] In some embodiments, the use of a 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a protein kinase inhibitor is a CDK4 inhibitor.

[0029] The present invention provides the use of the above-mentioned 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof in the preparation of drugs for treating or preventing protein kinase-mediated diseases, wherein the protein kinases include CDK4 and CDK6.

[0030] In some embodiments, the use of a 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a drug for treating or preventing a protein kinase-mediated disease, wherein the protein kinase is CDK4.

[0031] In some embodiments, the use of a 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a medicament for treating or preventing a protein kinase-mediated disease, wherein the protein kinase-mediated disease is cancer, and the cancer includes at least one of breast cancer, non-small cell lung cancer, colon cancer, prostate cancer, thyroid cancer, malignant melanoma, neuroblastoma, and mammary secretory carcinoma.

[0032] The present invention provides a pharmaceutical composition comprising:

[0033] Active ingredients and pharmaceutically acceptable carriers, wherein the active ingredients include the above-mentioned 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0034] Compared with the prior art, the 2-aminopyrimidine compounds of the present invention have the following beneficial effects:

[0035] The 2-aminopyrimidine compounds of the present invention are a series of novel compounds that can inhibit the activities of CDK4 and CDK6, especially with high selectivity, and can effectively treat or prevent cancer and other hyperproliferative diseases mediated by CDK4 and CDK6, especially cancer and other hyperproliferative diseases mediated by CDK4. DETAILED DESCRIPTION

[0036] The technical solutions of the present invention are further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. It should be understood that the purpose of providing these embodiments is to provide a more thorough and comprehensive understanding of the disclosure of the present invention.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0038] Definition of terms

[0039] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0040] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0041] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as formic acid, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0042] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0043] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0044] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0045] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0046] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0047] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond and straight dashed key

[0048] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is considered as a substituent to which it is connected), if a wavy line is used between the atom on the double bond and its substituent in the compound, When connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of two isomers of formula (A-1) and formula (A-2); the following formula (B) represents that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of two isomers of formula (B-1) and formula (B-2). The following formula (C) represents that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of two isomers of formula (C-1) and formula (C-2).

[0049] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and readily interconvert into each other. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0050] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0051] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0052] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0053] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. The type and number of substituents can be any chemically feasible basis. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.

[0054] When any variable (e.g., R1) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 instances of R1, the group may optionally be substituted with up to two instances of R1, with each instance of R1 being an independent choice. Furthermore, combinations of substituents and / or their variants are permitted only if such combinations result in stable compounds.

[0055] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0056] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0057] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the substituent is listed without specifying which atom it is connected to the substituted group, the substituent can be bonded through any atom of the substituent. For example, a pyridyl substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0058] When the listed linking groups do not specify their linking direction, their linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0059] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0060] Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, a "3-7 membered ring" refers to a "ring" having 3-7 atoms arranged around it.

[0061] Unless otherwise specified, the term “C 1-4 "Alkyl" is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. It can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-4 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, tert-butyl, and sec-butyl).

[0062] Unless otherwise specified, the term “C 1-4 "Alkoxy" means an alkyl group containing 1 to 4 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 1-3 Alkoxy. C 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0063] Unless otherwise specified, the term "alkylsulfonyl" refers to a -SO2R group where R is alkyl as defined above, for example, methylsulfonyl, ethylsulfonyl, and the like.

[0064]

[0046] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0065] Unless otherwise specified, the terms "5-membered heteroaryl ring" and "5-membered heteroaryl" are used interchangeably in the present invention. The term "5-membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). The 5-membered heteroaryl group may be attached to the rest of the molecule via a heteroatom or carbon atom. Examples of the 5-membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2 (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.).

[0066] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0067] Unless otherwise specified, “C 3-7 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 7 carbon atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. 3-7 Cycloalkyl groups include C 3-6 、C 4-6 、C 4-5 、C 5-7 or C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-7 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

[0068] Unless otherwise specified, the term "3-7 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 7 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). This includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings. In addition, with respect to the term "3-7 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The term "3-7 membered heterocycloalkyl" includes 5-7 membered, 3 membered, 4 membered, 5 membered, 6 membered and 7 membered heterocycloalkyls, etc. Examples of 3-7 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, etc.

[0069] Unless otherwise specified, the term "3-7 membered nitrogen-containing heterocycloalkyl" refers to a 3-7 membered heterocycloalkyl group containing at least one nitrogen atom.

[0070] Alicyclic refers to a saturated or partially unsaturated all-carbon ring system. Wherein "partially unsaturated" refers to a ring portion that includes at least one double bond or triple bond. "Partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein. Non-limiting examples include cyclopropyl rings, cyclobutyl rings, cyclopentyl rings, cyclopentenyl rings, cyclohexyl rings, cyclohexenyl rings, cyclohexadienyl rings, cycloheptyl rings, cycloheptatrienyl rings, cyclopentanone rings, cyclopentane-1,3-dione rings, and the like.

[0071] "Aryl" and "aromatic ring" are used interchangeably to refer to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, which may be fused to a cycloalkyl ring, a heterocycloalkyl ring, a cycloalkenyl ring, a heterocycloalkenyl ring, or a heteroaryl ring. 6-10 The term "aryl" refers to a monocyclic or bicyclic aromatic group having 6 to 10 carbon atoms. Non-limiting examples of aryl include phenyl, naphthyl, and the like.

[0072] "Heteroaryl" and "heteroaryl ring" are used interchangeably and refer to a group of a monocyclic, bicyclic or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having ring carbon atoms and ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In the present invention, heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl rings, heterocycloalkyl rings, cycloalkenyl rings, heterocycloalkenyl rings or aromatic rings. The heteroaryl ring may be optionally substituted. "5 to 10 membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl group having 5 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include thienyl, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and tetrazinyl. "8- to 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 8 to 10 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, The term "heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings.

[0073] The term "protecting group" includes, but is not limited to, an "amino protecting group," a "hydroxy protecting group," or a "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions at the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0074] Unless otherwise defined, the "substituents independently selected from..." described in the present invention means that when more than one hydrogen on a group is replaced by a substituent, the substituents may be the same or different, and the substituents selected are independently of each other.

[0075] Generally, the compound of the present invention or its pharmaceutically acceptable salt, or its stereoisomer can be formed into a suitable dosage form for administration with one or more pharmaceutical carriers. These dosage forms are suitable for oral, rectal, topical, oral and other parenteral administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules and syrups. The compound of the present invention contained in these preparations can be a solid powder or granules; a solution or suspension in an aqueous or non-aqueous liquid; an oil-in-water or water-in-oil emulsion, etc. The above dosage forms can be made from the active compound and one or more carriers or excipients through common pharmaceutical methods. The above carriers need to be compatible with the active compound or other excipients. For solid preparations, commonly used non-toxic carriers include but are not limited to mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, etc. Carriers for liquid preparations include water, physiological saline, aqueous glucose solution, ethylene glycol and polyethylene glycol, etc. The active compound can form a solution or suspension with the above carriers.

[0076] The compositions of the present invention are formulated, dosed and administered in a manner consistent with standard medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0077] "Therapeutically effective amount" refers to the amount of the compound of the present invention that will elicit a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0078] The therapeutically effective amount of the compound of the present invention or its pharmaceutically acceptable salt, or its stereoisomer contained in the pharmaceutical composition or pharmaceutical use composition of the present invention is preferably 0.1 mg-5 g / kg (body weight).

[0079] "Treatment" refers to alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0080] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0081] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0082] The solvents used in the present invention can be obtained commercially. The present invention uses the following abbreviations: Pd / C represents palladium on carbon; H2 represents hydrogen; N2 represents nitrogen; mL represents milliliter; NaNO2 represents sodium nitrite; HCl represents; SnCl2.2H2O represents dihydrated tin dichloride; H2O represents water; AcOH represents acetic acid; B2Pin2 represents bispinacol borate; Pd(dppf)Cl2 represents 1,1'-bisdiphenylphosphinoferrocene palladium dichloride; KOAc represents potassium acetate; EtOAc represents ethyl acetate; Pd(PPh3)4 represents tetrakis(triphenylphosphine)palladium; K2CO3 represents potassium carbonate; MeCN represents acetonitrile; DIEA represents N,N-diisopropylethylamine; MeOH represents methanol; NaBH3CN represents sodium cyanoborohydride; THF represents tetrahydrofuran; Boc2O represents di-tert-butyl carbonic anhydride; DMAP represents 4-dimethylaminopyridine; HCl / EtOAc represents ethyl acetate hydrochloride; MnO2 represents manganese dioxide; Cbz-Cl represents benzyl chloroformate; MsCl represents methanesulfonyl chloride; Pd(Amphos)2Cl2 represents dichlorodi-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II); SO2Cl2 represents dichlorosulfone; and NaHCO3 represents sodium bicarbonate.

[0083] Compounds are named according to the conventional nomenclature in the art or using The software named the commercially available compounds using the supplier's catalog name.

[0084] One embodiment of the present invention provides a 2-aminopyrimidine compound having a general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0085] in:

[0086] R1 is independently selected from halogen, cyano (-CN), trihalomethyl, nitro, carboxyl or acyl, and n1 is selected from 1 or 2;

[0087] R2, R4, and R5 are independently selected from H, halogen, C1-C4 alkyl, and C1-C4 alkoxy, n2 is selected from 1 or 2, n4 is selected from an integer of 1 to 4, and n5 is selected from an integer of 1 to 3;

[0088] R3 is selected from H, C1-C4 alkyl, C1-C4 alkoxy, (dialkylamino)methyl, acyl, hydroxy-substituted alkyl;

[0089] X represents O or NR6;

[0090] R6 is selected from alkylsulfonyl;

[0091] Y represents N or CR7;

[0092] R7 is selected from halogen, cyano, trihalomethyl, nitro, carboxyl, and acyl.

[0093] It can be understood that, as shown in the structure of formula (IA), in the general formula (I), the connection relationship between Ar1 and Ar2 can be any site of the 4th, 5th, and 6th positions of Ar1 connected to any site of the 1st, 2nd, and 6th positions of Ar2; R1 can appear at any site of the 4th, 5th, and 6th positions of Ar1 except the site connected to Ar2, and R2 can appear at any site of the 1st, 2nd, and 6th positions of Ar2 except the site connected to Ar1.

[0094] It is further understood that halogen includes but is not limited to F, Cl, Br; trihalomethyl includes but is not limited to trifluoromethyl, trichloromethyl, tribromomethyl; acyl includes but is not limited to formyl, acetyl, propionyl; C1-C4 alkyl includes but is not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl; C1-C4 alkoxy includes but is not limited to methoxy, ethoxy, propoxy; (dialkylamino)methyl includes but is not limited to (dimethylamino)methyl, (diethylamino)methyl; alkyl substituted with hydroxyl includes but is not limited to Not limited to hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 2-hydroxyisopropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 5-hydroxypentyl, 2-hydroxypentyl, 3-hydroxypentyl, 4-hydroxypentyl, 6-hydroxyhexyl, 2-hydroxyhexyl, 3-hydroxyhexyl, 4-hydroxyhexyl, 1-methyl-2-hydroxyethyl, 1,1-dimethyl-2-hydroxyethyl, 1,2-dihydroxyethyl, 2,2-dihydroxyethyl; alkylsulfonyl includes but is not limited to methylsulfonyl and ethylsulfonyl.

[0095] It can also be understood that n1 is selected from 1 or 2, which means that it may contain one or two R1s; n2 is selected from 1 or 2, which means that it may contain one or two R2s; n4 is selected from an integer from 1 to 4, and n4 is selected from 1, 2, 3 or 4, which means that it may contain 1, 2, 3 or 4 R4s; n5 is selected from an integer from 1 to 3, and n3 is selected from 1, 2 or 3, which means that it may contain 1, 2 or 3 R5s.

[0096] Currently, approved CDK4 / 6 inhibitors include Pfizer's Palbociclib, Novartis' LEE011, and Eli Lilly's Abemaciclib, used in combination with endocrine therapy for hormone receptor-positive, human epidermal growth factor receptor 2-negative advanced or metastatic breast cancer. However, treatment with CDK4 / 6 inhibitors has been shown to cause adverse reactions in clinical practice, such as gastrointestinal and hematologic toxicities.

[0097] Studies have found that CDK4 is a single oncogenic driver in many breast cancers.

[0098] Studies have shown that 2-aminopyrimidine compounds with the general formula (I) can inhibit the activities of CDK4 and CDK6, especially with high selectivity, and can effectively treat or prevent cancer and other hyperproliferative diseases mediated by CDK4 and CDK6, especially cancer and other hyperproliferative diseases mediated by CDK4.

[0099] The 2-aminopyrimidine compounds of the general formula (I) have a highly selective inhibitory effect on CDK4 activity and can effectively reduce the side effects caused by CDK4 / 6 inhibitors.

[0100] In some examples, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the general formula (I) is the structure of formula (I-1):

[0101] Furthermore, when n1, n2, n4, and n5 are all 1, the general formula (I) is the structure of formula (I-2):

[0102] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R1 is selected from halogen and cyano.

[0103] Furthermore, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R1 is selected from Cl and cyano.

[0104] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R3 is selected from C1-C4 alkyl, (dialkylamino)methyl, acyl, or hydroxy-substituted alkyl.

[0105] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R3 is selected from methyl, (dimethylamino)methyl Acetyl, 2-hydroxyisopropyl

[0106] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R2, R4, and R5 are independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0107] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R2 is H.

[0108] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R4 is H.

[0109] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, R5 is H.

[0110] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, X represents O or NR6; and R6 is selected from methylsulfonyl and ethylsulfonyl.

[0111] Further, R6 is methylsulfonyl; in this case NR6 is

[0112] In some examples, in the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, Y represents N or CR7; and R7 is selected from halogen and cyano.

[0113] Furthermore, R7 is selected from F and cyano.

[0114] In some examples, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the general formula (I) is selected from any one of formulas (II-1) to (II-9):

[0115] Accordingly, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is selected from the following compounds:

[0116] (3S,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol,

[0117] 1-(5'-(5-chloro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7'-fluorospiro[cyclopentane-1,3'-indol]-2'-yl)ethan-1-one,

[0118] (3S,4R)-4-((5-chloro-4-(2'-((dimethylamino)methyl)-7'-fluorospiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol,

[0119] (3S,4R)-4-((5-chloro-4-(7'-fluoro-2'-(2-hydroxypropan-2-ylspiro[cyclopentane-1,3'-indol]-5'-yl))pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol,

[0120] 4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indole]-5'-yl)-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carbonitrile,

[0121] 5'-(5-cyano-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-2'-methylspiro[cyclopentane-1,3'-indole]-7'-carbonitrile,

[0122] (3R,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol,

[0123] 4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indole]-5'-yl)-2-(((3R,4R)-3-hydroxy-1-(methylsulfonyl)piperidin-4-yl)amino)pyrimidine-5-carbonitrile,

[0124] (3S,4R)-4-((5-chloro-4-(2'-methylspiro[cyclopentane-1,3'-pyrrolo[2,3-b]pyridine]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol.

[0125] In some examples, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is selected from the following compounds:

[0126] (3S,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol,

[0127] 1-(5'-(5-chloro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7'-fluorospiro[cyclopentane-1,3'-indol]-2'-yl)ethan-1-one、

[0128] (3S,4R)-4-((5-chloro-4-(2'-((dimethylamino)methyl)-7'-fluorospiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol、

[0129] (3S,4R)-4-((5-chloro-4-(7'-fluoro-2'-(2-hydroxypropan-2-yl)spiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol、

[0130] 4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carbonitrile、

[0131] 5'-(5-cyano-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-2'-methylspiro[cyclopentane-1,3'-indole]-7'-carbonitrile、

[0132] (3R,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol、

[0133] 4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)-2-(((3R,4R)-3-hydroxy-1-(methylsulfonyl)piperidin-4-yl)amino)pyrimidine-5-carbonitrile,

[0134] (3S,4R)-4-((5-chloro-4-(2'-methylspiro[cyclopentane-1,3'-pyrrolo[2,3-b]pyridin]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol.

[0135] In some specific examples, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the general formula (I) is selected from any one of formulas (II-1), (II-2), (II-5), and (II-7):

[0136] In some examples, the 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the pharmaceutically acceptable salt is an alkyl acid salt.

[0137] Furthermore, the pharmaceutically acceptable salt is formate.

[0138] One embodiment of the present invention provides the use of the above-mentioned 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a protein kinase inhibitor, wherein the protein kinase inhibitor includes a CDK inhibitor.

[0139] In some examples, 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof are used in the preparation of protein kinase inhibitors, including CDK4 inhibitors and CDK6 inhibitors.

[0140] In some examples, a 2-aminopyrimidine compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof is used in the preparation of a protein kinase inhibitor, wherein the protein kinase inhibitor is a CDK4 inhibitor.

[0141] One embodiment of the present invention provides the use of the above-mentioned 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof in the preparation of drugs for treating or preventing protein kinase-mediated diseases, wherein protein kinase inhibitors include CDK inhibitors.

[0142] In some examples, 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof are used in the preparation of drugs for treating or preventing protein kinase-mediated diseases, wherein the protein kinases include CDK4 and CDK6.

[0143] In some examples, in the use of a 2-aminopyrimidine compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof for preparing a medicament for treating or preventing a protein kinase-mediated disease, the protein kinase is CDK4.

[0144] In some examples, 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof are used in the preparation of drugs for treating or preventing protein kinase-mediated diseases, wherein the protein kinase-mediated disease is cancer, and the cancer includes at least one of breast cancer, non-small cell lung cancer, colon cancer, prostate cancer, thyroid cancer, malignant melanoma, neuroblastoma and mammary secretory carcinoma.

[0145] In some examples, 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof are used in the preparation of drugs for treating or preventing protein kinase-mediated diseases, wherein the drugs for treating or preventing protein kinase-mediated diseases are drugs for inhibiting breast cancer cell proliferation.

[0146] Furthermore, the breast cancer cells are MCF7 cells or T47D cells.

[0147] One embodiment of the present invention provides a pharmaceutical composition comprising:

[0148] Active ingredients and pharmaceutically acceptable carriers, the active ingredients include the above-mentioned 2-aminopyrimidine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0149] It will be appreciated that pharmaceutically acceptable carriers include, but are not limited to, sustained-release agents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, adsorption carriers, surfactants, and lubricants.

[0150] Specific embodiments

[0151] The following examples are given based on the 2-aminopyrimidine compounds, applications thereof, and pharmaceutical compositions of the present invention. It should be understood that the 2-aminopyrimidine compounds, applications thereof, and pharmaceutical compositions of the present invention are not limited to the following examples.

[0152] Example 1

[0153] (3S,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (II-1)

[0154] The synthetic route is as follows:

[0155] Step 1:

[0156] NaNO2 (1.90 g, 27.5 mmol, 1.05 eq) was added dropwise to a solution of compound 1-1 (5.00 g, 26.3 mmol, 1.00 eq) in HCl (25.0 mL) at -20°C and stirred for 30 minutes. The mixture was then added dropwise to a solution of SnCl2.2H2O (19.0 g, 84.2 mmol, 3.20 eq) in HCl (25.0 mL) at -20°C. The resulting mixture was stirred at 0°C for 1.5 hours. The reaction system was diluted with 10.0 mL of H2O and extracted three times with 15.0 mL (5.00 mL*3) of ethyl acetate. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 1-2. LCMS: MS (ESI) m / z = 206.8 [M+H]+. 1 H NMR: CDCl3, 400MHz δ = 7.11 (dd, J = 1.2, 8.6Hz, 1H), 7.04 (dd, J = 2.0, 11.0Hz, 1H), 6.94 (t, J = 8.8Hz, 1H), 5.34 (br s, 1H), 3.48 (br s, 2H).

[0157] Step 2:

[0158] At room temperature, compound 1-2 (2.00 g, 9.75 mmol, 1.00 eq) and compound 1-3 (1.20 g, 10.7 mmol, 1.10 eq) were added to a 10.0 mL solution of AcOH and stirred at 120°C for 5 hours. 10.0 mL of H2O was added to the reaction system for dilution, and the mixture was extracted three times with 45.0 mL (15.0 mL*3) of ethyl acetate. The mixture was then washed three times with 45.0 mL (15.0 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 1-4. LCMS: MS (ESI) m / z = 283.7 [M+H]+. 1 H NMR: CDCl3, 400MHz δ = 7.15 (d, J = 1.6 Hz, 1H), 7.12 (dd, J = 1.6, 9.0 Hz, 1H), 2.23 (s, 3H), 1.99-1.90 (m, 6H), 1.76-1.70 (m, 2H).

[0159] Step 3:

[0160] At room temperature, compound 1-4 (500 mg, 1.77 mmol, 1.00 eq), B2Pin2 (585 mg, 2.30 mmol, 1.30 eq), Pd(dppf)Cl2 (144 mg, 177 umol, 0.10 eq), and KOAc (521 mg, 5.32 mmol, 3.00 eq) were added to a dioxane (5.00 mL) solution, replaced with gas three times under N2 atmosphere, and then stirred at 90 ° C for 12 hours; 10.0 mL of H2O was added to the reaction system for dilution, extracted three times with EtOAc 15.0 mL (5.00 mL * 3), and then washed three times with saturated sodium chloride solution 15.0 mL (5.00 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product compound 1-5 (500 mg) as a white solid. LCMS: MS (ESI) m / z=329.8[M+H]+. 1 H NMR: CDCl3, 400MHz δ = 7.41 (s, 1H), 7.38 (d, J = 10.4Hz, 1H), 2.26 (s, 3H), 2.05 (br d,J=3.2Hz,2H),1.96-1.88(m,5H),1.82-1.75(m,2H),1.28(s,13H),1.17(s,6H).

[0161] Step 4:

[0162] At room temperature, compound 1-5 (400 mg, 1.22 mmol, 1.00 eq), compound 1-6 (222 mg, 1.22 mmol, 1.00 eq), Pd(PPh3)4 (140 mg, 121 umol, 0.100 eq), and K2CO3 (503 mg, 3.65 mmol, 3.00 eq) were added to a mixed solution of dioxane (3.00 mL) and water (1.00 mL). The mixture was replaced three times under a N2 atmosphere and then stirred at 90°C for 12 hours. 10.0 mL of H2O was added to the reaction system for dilution, and the mixture was extracted three times with 15.0 mL (5.00 mL*3) of ethyl acetate. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Compound 1-7 was purified by preparative HPLC (FA conditions) to obtain compound 1-7. LCMS: MS (ESI) m / z=349.8[M+H]+. 1H NMR: CDCl3, 400MHz δ = 8.58 (s, 1H), 7.61-7.55 (m, 2H), 2.31 (s, 3H), 2.06-1.96 (m, 6H), 1.86-1.78 (m, 2H).

[0163] Step 5:

[0164] At room temperature, compound 1-7 (400 mg, 1.14 mmol, 1.00 eq) and compound 1-8 (267 mg, 2.28 mmol, 2.00 eq) were dissolved in MeCN (4.00 mL). DIEA (147 mg, 1.14 mmol, 198 uL, 1.00 eq) was added and stirred at 80°C for 12 hours. The reaction system was diluted with 10.0 mL of H2O and extracted three times with 15.0 mL (5.00 mL*3) of ethyl acetate. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Compound II-1 was purified by preparative HPLC (FA conditions: Phenomenexluna C18 150*25.0 mm*10.0 um; mobile phase: [water (FA)-ACN]; B%: 36.0%-66.0%, 15.0 min). LCMS: MS (ESI) m / z = 432.1 [M+H]+. 1 H NMR: DMSO-d6, 400MHz δ = 8.41 (s, 1H), 7.64 (brdd, J = 1.6, 5.1Hz, 1H), 7.58-7. 46(m,2H),4.94-4.89(m,1H),3.94-3.70(m,4H),3.56-3.43(m,1H),3.04(br t,J=10.4Hz,1H),2.32(s,3H),2.06-1.91(m,7H),1.73(br d,J=4.0Hz,2H),1.55-1.45(m,1H).

[0165] Example 2

[0166] 1-(5'-(5-chloro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-7'-fluorospiro[cyclopentane-1,3'-indol]-2'-yl)ethan-1-one (II-2)

[0167] The synthetic route is as follows:

[0168] Step 1:

[0169] Compound 1-2 (5.00 g, 24.3 mmol, 1.00 eq) and compound 2-1 (4.15 g, 24.3 mmol, 1.00 eq) were added to a solution of AcOH (10.0 mL) at room temperature and stirred at 120°C for 5 hours. The reaction system was diluted with 300 mL of H2O and extracted three times with 300 mL of EtOAc (100 mL x 3). The mixture was then washed three times with 300 mL of saturated sodium chloride solution (100 mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to obtain compound 2-2. LCMS: MS (ESI) m / z = 342.0 [M+H]+. 1 H NMR: CDCl3, 400MHz δ = 7.34-7.30 (m, 1H), 7.28 (s, 1H), 4.46 (q, J = 7.2Hz, 2H), 2.57 -2.45(m,2H),2.27-2.01(m,5H),1.79(td,J=6.1,12.2Hz,3H),1.49-1.42(m,3H).

[0170] Step 2:

[0171] Compound 2-2 (1.00 g, 2.94 mmol, 1.00 eq) was added to a mixture of AcOH (6.00 mL) and MeOH (8.00 mL) at room temperature, followed by the addition of NaBH3CN (554 mg, 8.82 mmol, 3.00 eq). The mixture was stirred at 25°C for 5 hours. The reaction system was diluted with 30.0 mL of H2O and extracted three times with 30.0 mL (10.0 mL*3) of EtOAc. The mixture was then washed three times with 30.0 mL (10.0 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, Compound 2-3. LCMS: MS (ESI) m / z = 343.7 [M+H]+.

[0172] Step 3:

[0173] To a solution of compound 2-3 (1.00 g, 2.92 mmol, 1.00 eq) in THF (1.00 mL) at room temperature, Boc2O (6.38 g, 29.2 mmol, 6.71 mL, 10.0 eq) and DMAP (35.7 mg, 292 μmol, 0.100 eq) were added, and the mixture was stirred at 25°C for 4 hours. The reaction system was diluted with 30.0 mL of H2O and extracted three times with 30.0 mL (10.0 mL*3) of EtOAc. The mixture was then washed three times with 30.0 mL (10.0 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, compound 2-4. LCMS: MS (ESI) m / z = 341.8 [M+H]+. 1 H NMR: CDCl3, 400MHz δ = 7.07-7.00 (m, 1H), 6.88 (d, J = 1.8Hz, 1H), 4.53 (s, 1H), 4.14-4. 06(m,2H),1.82-1.76(m,4H),1.73-1.54(m,4H),1.44(s,10H),1.18(d,J=7.1Hz,3H).

[0174] Step 4:

[0175] To a solution of compound 2-4 (400 mg, 904 μmol, 1.00 eq) in THF (4.00 mL) at -20°C, compound 2-5 (185 mg, 1.90 mmol, 2.10 eq) was added, followed by the slow dropwise addition of isopropylmagnesium chloride (2.00 M, 2.03 mL, 4.50 eq). The mixture was stirred at -20°C for 1 hour. The reaction system was diluted with 30.0 mL of H2O at 0°C and extracted three times with 30.0 mL (10.0 mL*3) of EtOAc. The mixture was then washed three times with 30.0 mL (10.0 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, compound 2-6. LCMS: MS (ESI) m / z = 341.9 [M+H]+.

[0176] Step 5:

[0177] Methylmagnesium bromide (2.50 M, 1.05 mL, 3.00 eq) was slowly added dropwise to a solution of compound 2-6 (400 mg, 874 μmol, 1.00 eq) in THF (4.00 mL) at -30°C, and the mixture was stirred at -30°C for 1 hour. The reaction system was diluted with 10.0 mL of H2O at 0°C, extracted three times with 9.00 mL (3.00 mL*3) of EtOAc, and then washed three times with 9.00 mL (3.00 mL*3) of saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 2-7. LCMS: MS (ESI) m / z = 314.0 [M+H]+.

[0178] Step 6:

[0179] Compound 2-7 (300 mg, 727 μmol, 1.00 eq) was dissolved in HCl / EtOAc (3.00 mL) at room temperature and stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain the crude product, Compound 2-8. LCMS: MS (ESI) m / z = 313.8 [M+H]+.

[0180] Step 7:

[0181] MnO2 (306 mg, 3.52 mmol, 5.00 eq) was added to a solution of compound 2-8 (220 mg, 704 μmol, 1.00 eq) in dichloromethane (2.00 mL) at room temperature, and the mixture was stirred at 25°C for 3 hours. The reaction system was diluted with 10.0 mL of H2O at 0°C, extracted three times with 9.00 mL (3.00 mL*3) of EtOAc, and then washed three times with 9.00 mL (3.00 mL*3) of saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 2-9. LCMS: MS (ESI) m / z = 309.8 [M+H]+. 1 H NMR: CDCl3, 400MHz δ = 4.10-4.00 (m, 1H), 2.60 (s, 3H), 2.37-2.27 (m, 2H), 2.21-2.08 (m, 2H), 1.99-1.92 (m, 3H), 1.67-1.57 (m, 3H).

[0182] Step 8:

[0183] Compound 2-9 (20.0 mg, 64.4 μmol, 1.00 eq) and B2Pin2 (81.8 mg, 322 μmol, 5.00 eq) were dissolved in dioxane (0.500 mL) at room temperature. Pd(dppf)Cl2 (4.72 mg, 6.45 μmol, 0.10 eq) and KOAc (18.9 mg, 193 μmol, 3.0 eq) were then added and stirred at 80°C for 12 hours. The reaction mixture was diluted with 10.0 mL of H2O at 0°C and extracted three times with 9.00 mL (3.00 mL*3) of EtOAc. The mixture was then washed three times with 9.00 mL (3.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, Compound 2-10. LCMS: MS (ESI) m / z = 358.0 [M+H]+.

[0184] Step 9:

[0185] At room temperature, compound 2-10 (23.0 mg, 64.3 umol, 1.00 eq) and compound 1-6 (11.8 mg, 64.3 umol, 1.0 eq) were dissolved in a mixture of dioxane (0.500 mL) and water (0.200 mL). Pd(PPh3)4 (7.44 mg, 6.44 umol, 0.10 eq) and K2CO3 (26.7 mg, 193 umol, 3.00 eq) were then added and stirred at 80°C for 3 hours. 10.0 mL of H2O was added to the reaction system at 0°C, and the mixture was extracted three times with 9.00 mL (3.00 mL*3) of EtOAc, then washed three times with 9.00 mL (3.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The product was purified by preparative high performance liquid chromatography (FA conditions) to obtain compound 2-11. LCMS: MS (ESI) m / z = 335.8 [M+H] +.

[0186] Step 10:

[0187] At room temperature, compound 2-11 (13.0 g, 34.3 μmol, 1.00 eq) and compound 1-8 (8.05 mg, 68.7 μmol, 2.00 eq) were dissolved in MeCN (0.500 mL). DIEA (13.3 mg, 103 μmol, 17.9 μL, 3.00 eq) was added and stirred at 80°C for 12 hours. 10.0 mL of H2O was added to the reaction system at 0°C and the mixture was extracted three times with 9.00 mL (3.00 mL*3) of EtOAc. The mixture was then washed three times with 9.00 mL (3.00 mL*3) of saturated sodium chloride solution. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The product was purified by preparative HPLC (neutral column: Waters xbridge 150*25.0 mm 10 μm; mobile phase: [water(NH 4 HCO 3 )-ACN]; B%: 42%-72%, 8.0 min) to obtain compound 2. LCMS: MS (ESI) m / z=459.0 [M+H] + . 1 H NMR: DMSO-d6, 400MHz δ = 8.44 (s, 1H), 7.81-7.52 (m, 3H), 4.93 (d, J = 5.6Hz, 1H), 3.90-3.78 (m, 3H), 3.57-3.44 (m, 2H), 3.03 (br t,J=10.4Hz,1H),2.62(s,3H),2.42-2.30(m,2H),2.19-1.86(m,6H),1.70(brdd,J=5.8,11.3Hz,2H).

[0188] Example 3

[0189] (3R,4R)-4-((5-chloro-4-(7'-fluoro-2'-methylspiro[cyclopentane-1,3'-indol]-5'-yl)pyrimidin-2-yl)amino)-1-(methylsulfonyl)piperidin-3-ol (II-7)

[0190] The synthetic route is as follows:

[0191] Step 1:

[0192] Compound 3-1 (500 mg, 2.31 mmol, 1.00 eq) was dissolved in dichloromethane (5.00 mL) and saturated aqueous sodium carbonate (5.00 mL) at room temperature. Cbz-Cl (473 mg, 2.77 mmol, 394 uL, 1.20 eq) was added dropwise at 0°C. After the addition was complete, the mixture was reacted at 0°C for 3 hours and monitored. The reaction solution was slowly poured into water (5.00 mL) at 0°C and extracted three times with dichloromethane (5.00 mL x 3). The organic phases were combined, washed with saturated aqueous sodium chloride (5.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 50:1 to 3:1, product: Rf = 0.50) to obtain compound 3-2. LCMS: m / z = 251.1 [M+H]+. 1 H NMR: (400MHz DMSO-d6)δ7.37-7.28(m,4H),7.15(br d,J=8.0Hz,1H),5.01(s,2H),3.76(br d,J=13.2Hz,1H),3.31-3.26(m,1H),3.20(tt,J=4.8,9.2Hz,1H),1.81-1.69(m,1H),1.38(s,9H),1.29-1.19(m,4H).

[0193] Step 2:

[0194] Compound 3-2 (770 mg, 2.20 mmol, 1.00 eq) was dissolved in dichloromethane (5.00 mL) at room temperature, and HCl / EtOAc (4.00 M, 5.50 mL, 10.0 eq) was added to the mixture. The reaction was allowed to react at 25°C for 2 hours. The reaction solution was directly concentrated to obtain the hydrochloride salt of compound 3-3. LCMS: m / z = 251.0 [M+H]+. 1 H NMR:(400MHz DMSO-d6)δ8.42(br s,1H),7.54-7.41(m,1H),7.38-7.28(m,4H),5.09-4.97(m,2H),3.65(td,J=4.0,7.6Hz,1H),3.55-3.46(m,1H),3.21-3.09(m,2H),2.90(br d,J=6.4Hz,1H),2.81-2.71(m,1H),2.07-1.94(m,1H),1.69-1.51(m,1H).

[0195] Step 3:

[0196] At room temperature, compound 3-3 (150 mg, 523 μmol, 1.00 eq, HCl) was dissolved in ethyl acetate (0.50 mL) and saturated aqueous sodium bicarbonate (0.50 mL). After replacing the nitrogen atmosphere three times, MsCl (180 mg, 1.57 mmol, 121 μL, 3.00 eq) was added dropwise to the reaction solution at 0°C. After the addition was complete, the mixture was reacted at 0°C for 2 hours. The reaction solution was slowly poured into water (1.00 mL) at 0°C and extracted three times with ethyl acetate (1.00 mL x 3). The organic phases were combined, washed with saturated aqueous sodium chloride (1.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain the crude product. The crude product was purified by preparative HPLC (preparative column: Welch Ultimate C18 150*25.0 mm*5.0 um; mobile phase: [water (TFA)-ACN]; B%: 16.0%-46.0%, 10.0 min) and lyophilized to obtain compound 3-4. LCMS: m / z = 329.1 [M+H] + . 1 H NMR: (400MHz DMSO-d6)δ7.38-7.31(m,4H),7.25(br d,J=8.4Hz,1H),5.18(br d,J=3.2Hz,1H),5.02(s,2H),3.56-3.48(m,1H),3.45-3.35(m,2H),3.31-3.24(m,1H),2.86(s,3H) ,2.80(dt,J=2.4,11.7Hz,1H),2.59(dd,J=9.2,11.2Hz,1H),1.93-1.83(m,1H),1.50-1.36(m,1H).

[0197] Step 4:

[0198] At room temperature, compound 3-4 (80.0 mg, 243 μmol, 1.00 eq) was dissolved in dichloromethane (1.00 mL) and methanol (0.80 mL). The air was replaced with H₂ three times, and then Pd / C (20.0 mg, 487 μmol, 10% purity, 2.00 eq) was added. The air was replaced with H₂ three times, and the mixture was reacted at 20°C under 15 psi of H₂ for 1 hour. The reaction solution was filtered, and the filtrate was concentrated to give compound 3-5. LCMS: EC6457-348-P1B1, product: RT = 0.124 min, m / z = 195.1 [M+H]⁺.

[0199] Step 5:

[0200] To a solution of compound 1-7 (20.0 mg, 57.1 μmol, 1.00 eq) and compound 3-5 (20.0 mg, 102 μmol, 1.80 eq) in MeCN (0.50 mL) was added DIEA (22.1 mg, 171 μmol, 29.8 μL, 3.00 eq) at room temperature. The mixture was stirred at 80°C for 72 hours. The reaction solution was slowly poured into water (1.00 mL) and extracted three times with ethyl acetate (1.00 mL x 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (1.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain the crude product. The crude product was purified by preparative HPLC (preparative column: Welch Ultimate C18 150*25.0 mm*5.0 um; mobile phase: [water (TFA)-ACN]; B%: 30%-60%, 10.0 min) and lyophilized to obtain compound 3. LCMS: m / z = 508.3 [M+H] + . 1 H NMR:(400MHz DMSO-d6)δ8.41(s,1H),7.71-7.59(m,1H),7.58-7.42(m,2H),5.30-5.09(m,1H),3.86-3.73(m,1H),3.67-3.55(m,2H) ,3.51-3.45(m,1H),2.89(s,3H),2.87-2.79(m,1H),2.69-2.60(m,1H),2.35-2.27(m,3H),2.10-1.95(m,7H),1.73(br dd,J=4.5,6.8Hz,2H),1.59-1.45(m,1H).

[0201] Example 4

[0202] 4-(7'-Fluoro-2'-methylspiro[cyclopentane-1,3'-indole]-5'-yl)-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carbonitrile (II-5)

[0203] The synthetic route is as follows:

[0204] Step 1:

[0205] At room temperature, compound 1-5 ((200 mg, 607 umol, 1.00 eq), compound 4-1 (112 mg, 607 umol, 1.00 eq), K2CO3 (251 mg, 1.82 mmol, 3.00 eq) and Pd(Amphos)2Cl2 (43.0 mg, 60.7 umol, 43.0 uL, 0.100 eq) were dissolved in dioxane (2.00 mL) and The mixture was replaced with water (0.500 mL) and nitrogen was replaced three times. The reaction was continued at 90°C under a nitrogen atmosphere for 12 hours. Water (2.00 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (5.00 mL*3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (10.0 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4-2. LCMS: MS (ESI) m / z = 330.1 [M+H]+.

[0206] Step 2:

[0207] At 0°C, compound 4-2 (85.0 mg, 241 μmol, 1 eq) was dissolved in acetonitrile (0.500 mL), and SO2Cl2 (97.6 mg, 723 μmol, 72.3 μL, 3.00 eq) was slowly added dropwise. The mixture was stirred at 0°C for 1 hour. At 0°C, the reaction system was slowly added to NaHCO3 (aq, 2.00 mL) and extracted three times with ethyl acetate (2.00 mL*3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (5.00 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4-3. LCMS: m / z = 341.0 (M+H)+

[0208] Step 3:

[0209] Compound 4-3 (50.0 mg, 146 μmol, 1.00 eq), compound 4-4 (27.0 mg, 176 μmol, 1.20 eq), and DIEA (56.8 mg, 440 μmol, 76.6 μL, 3.00 eq) were dissolved in acetonitrile (0.500 mL) at room temperature and reacted at 80°C for 12 hours. Water (1.00 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (2.00 mL*3). The organic phases were combined, washed three times with saturated sodium chloride solution (4.00 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product, which was purified by preparative HPLC (FA conditions) to yield compound 4. LCMS: m / z = 422.2 [M+H]+. 1H NMR: (400MHz, DMSO-d6) δ8.81-8.72(m,1H),8.44-8.32(m,1H),7.93-7.81(m,1H),7.77-7.65(m,1H),5.02-4.96(m,1H),4.07-3.93(m,1H),3.82(br dd,J=4.8,10.7Hz,2H),3.58-3.47(m,1H),3.04(dt,J=4.4,10.4Hz,1H),2.33(s,3H),2.07(br d,J=3.6Hz,6H),1.98-1.83(m,1H),1.72(br d,J=5.2Hz,2H),1.61-1.20(m,2H).

[0210] In vitro activity test

[0211] Experimental Example 1: In vitro CDK4 / CyclinD1 enzyme activity test

[0212] Experimental Materials:

[0213] CDK4 / CyclinD1 was purchased from Invitrogen; Ulight-4E-BP1 peptide, 1X detection buffer, and Eu-anti-phospho-tyrosine antibody were purchased from PerkinElmer; high-purity ATP was purchased from Promega; EDTA was purchased from Sigma; and Nivo multilabel analyzer was used (PerkinElmer).

[0214] Experimental methods:

[0215] Kinase buffer contains: 50 mM HEPES, 1 mM EDTA, 10 mM MgCl2, 0.01% Brij-35, pH 7.4;

[0216] Prepare kinase buffer: Add 2.38 g HEPES, 58 mg EDTA, 406 mg MgCl2, and 20 mg Brij-35 to 200 mL of buffer and adjust the pH to 7.4.

[0217] Prepare stop solution: Use 100 μL of 1M EDTA stock solution, add 0.625 μL of 1X detection buffer and mix with 1725 μL of distilled water.

[0218] Enzyme, Ulight-4E-BP1 peptide, ATP, and inhibitor were diluted in kinase buffer. Eu-anti-phospho-tyrosine antibody was diluted to 8 nM / L in assay buffer. Test compounds were diluted five-fold using a pipette to an eighth concentration, from 8 μM to 0.102 nM, with a final DMSO concentration of 4%. A duplicate assay was performed. 2.5 μL of each inhibitor concentration gradient and 5 μL of CDK4 / CyclinD1 enzyme (0.5 ng) were added to the microplate. The compounds and enzyme were incubated at 25°C for 1 hour. 2.5 μL of a substrate and ATP mixture (4 mM ATP, 100 nM Ulight-4E-BP1 peptide) was added to the microplate. The final compound concentrations were then diluted from 2 μM to 0.0256 nM, with final ATP and substrate concentrations of 1 mM and 25 nM, respectively. The reaction system was incubated at 25°C for 180 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution was added to each well, and the reaction was carried out at 25°C for 60 minutes. Data were collected using the PerkinElmer Nivo multi-label analyzer in TR-FRET mode (excitation wavelength was 320 nm, emission wavelengths were 615 nm and 665 nm).

[0219] Experimental Example 2: In vitro CDK6 / CyclinD1 enzyme activity test

[0220] Experimental Materials:

[0221] CDK6 / CyclinD1 was purchased from Carna; Ulight-4E-BP1 peptide, Eu-anti-phospho-tyrosine antibody, and 1X detection buffer were purchased from PerkinElmer; high-purity ATP was purchased from Promega; EDTA was purchased from Sigma; and Nivo Multilabel Analyzer was used (PerkinElmer).

[0222] Experimental methods:

[0223] Kinase buffer contains: 50 mM HEPES, 1 mM EDTA, 10 mM MgCl2, 0.01% Brij-35, pH 7.4;

[0224] Prepare kinase buffer: Add 2.38 g HEPES, 58 mg EDTA, 406 mg MgCl2, and 20 mg Brij-35 to 200 mL of buffer and adjust the pH to 7.4.

[0225] Prepare stop solution: Use 100 μL of 1M EDTA stock solution, add 0.625 μL of 1X detection buffer and mix with 1725 μL of distilled water.

[0226] Dilute the enzyme, Ulight-4E-BP1 peptide, ATP, and inhibitor in kinase buffer. Dilute the Eu-anti-phospho-tyrosine antibody to 8 nM / L in assay buffer. Use a pipette to dilute the test compound five-fold to the eighth concentration, from 8 μM to 0.102 nM, with a final DMSO concentration of 4%. Set up a duplicate assay. Add 2.5 μL of each inhibitor concentration gradient, 5 μL of 2 ng of CDK6 / CyclinD1 enzyme, and 2.5 μL of a substrate and ATP mixture (4 mM ATP, 50 nM Ulight-4E-BP1 peptide) to the microplate. Final compound concentrations range from 2 μM to 0.0256 nM, with final ATP and substrate concentrations of 1 mM and 12.5 nM, respectively. Incubate the reaction at 25°C for 60 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution was added to each well, and the reaction was carried out at 25°C for 60 minutes. Data were collected using the PerkinElmer Nivo multi-label analyzer in TR-FRET mode (excitation wavelength was 320 nm, emission wavelengths were 615 nm and 665 nm).

[0227] Experimental Example 3: In vitro MCF-7 cell activity test

[0228] Experimental Materials:

[0229] EMEM culture medium was from Vicente; fetal bovine serum was from Biosera; penicillin / streptomycin antibiotics were purchased from Yuanpei; CellTiter-Glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega; MCF-7 cell line was purchased from Promega; Envision multi-label analyzer was purchased from PerkinElmer.

[0230] Experimental methods:

[0231] MCF-7 cells were seeded in a white 96-well plate, with 80 μL of cell suspension per well containing 2000 MCF-7 cells. The plate was cultured overnight in a CO2 incubator. The test compound was diluted 5-fold using a dispenser to the eighth concentration, from 2000 μM to 0.0256 nM, in duplicate. 78 μL of culture medium was added to the middle plate. 2 μL of the serially diluted compound was transferred to the middle plate according to the corresponding position. After mixing, 20 μL of the compound was transferred to each well of the plate. The concentration of the compound transferred to the plate ranged from 10 μM to 0.128 nM. The plate was cultured in a CO2 incubator for 7 days. A separate plate was prepared. The signal value was read on the day of drug addition and used as the maximum value (Max in the equation below) for data analysis. 25 μL of cell viability chemiluminescent detection reagent was added to each well of this plate and incubated at room temperature for 10 minutes to allow the luminescent signal to stabilize. The data were read using a multi-label analyzer. Add 25 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Read the results using a multi-label analyzer.

[0232] Experimental Example 4: In vitro T47D cell activity test

[0233] Experimental Materials:

[0234] 1640 culture medium was from Vivacell; fetal bovine serum was from Biosera; penicillin / streptomycin antibiotics were purchased from Yuanpei; CellTiter-Glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega; T-47D cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.; Envision multi-label analyzer was purchased from PerkinElmer.

[0235] Experimental methods:

[0236] T-47D cells were seeded in a white 384-well plate. 40 μL of cell suspension was added to each well, containing 300 T-47D cells. The plate was incubated overnight in a CO2 incubator. The test compound was diluted 5-fold using a dispenser to the eighth concentration, from 2000 μM to 0.0256 μM, in duplicate. 78 μL of culture medium was added to the middle plate. 2 μL of the serially diluted compound was transferred to the middle plate according to the corresponding position. After mixing, 10 μL of the compound was transferred to each well of the plate. The concentration of the compound transferred to the plate ranged from 10 μM to 0.128 nM. The plate was incubated in a CO2 incubator for 7 days. A separate plate was prepared. The signal value was read on the day of drug addition and used as the maximum value (Max in the equation below) for data analysis. 10 μL of cell viability chemiluminescent detection reagent was added to each well of this plate and incubated at room temperature for 10 minutes to allow the luminescent signal to stabilize. The data were read using a multilabel analyzer. Add 10 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Read the results using a multi-label analyzer.

[0237] Data Analysis:

[0238] The raw data of Experimental Examples 1 to 4 were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value of can be obtained by four-parameter curve fitting (obtained by log(inhibitor) vs.response--Variable slope mode in GraphPad Prism), and the results are shown in Table 1.

[0239] Table 1

[0240] As can be seen from Table 1, the compounds prepared in Examples 1 to 4 have different degrees of inhibitory effects on CDK4 and CDK6, especially highly selective inhibition of CDK4; and can effectively inhibit the proliferation of MCF7 cells and T47D cells.

[0241] The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0242] The embodiments described above only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in a specific and detailed manner, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present invention, several variations and improvements can be made, which all fall within the scope of protection of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the scope of protection of the claims attached to the present invention. Therefore, the scope of protection of the patent of the present invention shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. A 2-aminopyrimidine compound having the general formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in: R1 is independently selected from halogen, cyano, trihalomethyl, nitro, carboxyl or acyl, and n1 is selected from 1 or 2; R2, R4, and R5 are independently selected from H, halogen, C1-C4 alkyl, and C1-C4 alkoxy, n2 is selected from 1 or 2, n4 is selected from an integer of 1 to 4, and n5 is selected from an integer of 1 to 3; R3 is selected from H, C1-C4 alkyl, C1-C4 alkoxy, (dialkylamino)methyl, acyl, hydroxy-substituted alkyl; X represents O or NR6; R6 is selected from alkylsulfonyl; Y represents N or CR7; R7 is selected from halogen, cyano, trihalomethyl, nitro, carboxyl, and acyl.

2. The 2-aminopyrimidine compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The general formula (I) is the structure of formula (I-1):

3. The 2-aminopyrimidine compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The general formula (I) has at least one of the following characteristics (1) to (2): (1) R1 is selected from halogen and cyano; (2) R3 is selected from C1-C4 alkyl, (dialkylamino)methyl, acyl, and hydroxy-substituted alkyl.

4. The 2-aminopyrimidine compound according to claim 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The general formula (I) has at least one of the following characteristics (1) to (2): (1) R1 is selected from Cl, cyano; (2) R3 is selected from methyl, (dimethylamino)methyl, acetyl, and 2-hydroxyisopropyl.

5. The 2-aminopyrimidine compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The general formula (I) has at least one of the following characteristics (1) to (3): (1) R2 is H; (2) R4 is H; (3) R5 is H.

6. The 2-aminopyrimidine compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: X represents O or NR6; R6 is methylsulfonyl.

7. The 2-aminopyrimidine compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: Y represents N or CR7; R7 is selected from halogen and cyano.

8. The 2-aminopyrimidine compound according to claim 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R7 is selected from F and cyano.

9. The 2-aminopyrimidine compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: The general formula (I) is selected from any one of formulas (II-1) to (II-9):

10. Use of the 2-aminopyrimidine compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a protein kinase inhibitor, wherein the protein kinase inhibitor includes a CDK4 inhibitor and a CDK6 inhibitor. The use according to claim 10 , wherein the protein kinase inhibitor is a CDK4 inhibitor.

12. Use of the 2-aminopyrimidine compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a drug for treating or preventing protein kinase-mediated diseases, wherein the protein kinases include CDK4 and CDK6.

13. The use according to claim 12, wherein the protein kinase is CDK4.

14. The use according to claim 12, wherein the protein kinase-mediated disease is cancer, and the cancer comprises at least one of breast cancer, non-small cell lung cancer, colon cancer, prostate cancer, thyroid cancer, malignant melanoma, neuroblastoma and mammary secretory carcinoma.

15. A pharmaceutical composition, characterized in that include: An active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises the 2-aminopyrimidine compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.