WO2019170055A1 - Compounds having cdk4/6 kinase inhibitory activity, pharmaceutical composition thereof and use thereof - Google Patents

Abstract

The present invention relates to compounds having CDK4/6 kinase inhibitory activity, a pharmaceutical composition thereof and a use thereof. The present invention relates in particular to compounds represented by formula I, and further relates to a pharmaceutical composition comprising such compounds and a method for using such compounds in the prevention or treatment of diseases, especially for use with diseases mediated by CDK kinase, in particular by CDK4/6 kinase aberrant activity.

Description

Compound having CDK4/6 kinase inhibitory activity, pharmaceutical composition thereof and use thereof Technical field

The present invention relates to a compound having kinase inhibitory activity, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, and use thereof . The invention also relates to pharmaceutical compositions comprising these compounds and to methods of preventing or treating diseases, particularly those diseases mediated by CDK kinase, in particular CDK4/6 kinase aberrant activity.

Background technique

Abnormal cell cycle is a hallmark of cancer. Cyclin-dependent kinase (CDK) is a type of serine/threonine kinase that plays a central role in the cell cycle, leading to the initiation and progression of the cell cycle. End. The CDK family is an important signal transduction molecule in the cell, and its CDK-cyclin complex with cyclin is involved in cell growth, proliferation, dormancy and apoptosis.

In the past 20 years, drug development with CDK kinase as a therapeutic target for cancer has received extensive attention, such as Flavopiridol (Alvocidib), Seliciclib (CYC202), Dinaciclib (SCH727965) and Milciclib (PHA-848125). Clinical studies at different stages. However, due to the early detection of CDK inhibitors, the inhibition activity of each CDK family subtype is not high, or lack of certain selectivity, or poor absorption in vivo, which limits the clinical application. In recent years, drug discovery in this field has been made by increasing the selectivity of CDK inhibitors for each CDK family subtype or increasing the inhibitory activity of CDK kinase, especially the selective inhibitors targeting CDK4/6. Become a hot spot again.

CDK4/6 is overactive in many cancers, leading to uncontrolled cell proliferation. Thus, inhibition of CDK4/6 can achieve inhibition of cell proliferation downstream of the signaling pathway. At present, Pfizer's PD0332991 (Palbociclib) and Novartis's LEE-011 (Ribociclib) have been marketed as drugs for the treatment of ER-positive, HER2-negative breast cancer. LY-2835219 (abemaciclib), a similar drug, is expected to be available around 2017, and there are a large number of similar compounds in preclinical and early clinical research stages at different stages of development. However, there are some unsolved problems in these compounds, such as poor efficacy of monotherapy, low selectivity to other targets, and resistance to brain metastasis. Therefore, there is an urgent need in the field to research and develop new ones. A novel CDK kinase inhibitor with high efficacy, low toxicity, resistance to drug resistance and clinical application value.

Summary of the invention

In one aspect, the invention provides a compound of formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate thereof or Prodrug,

among them,

R _{1 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₂ -C ₆ alkenyl, optionally substituted C ₂ -C ₆ alkynyl, optionally substituted C ₃ - a C ₈ cycloalkyl group, an optionally substituted 4-8 membered heterocyclic group containing 1, 2 or 3 hetero atoms selected from nitrogen, oxygen and sulfur, an optionally substituted carbonyl group, an optionally substituted sulfonyl group, and An optionally substituted sulfinyl group;

R _{2 is} selected from the group consisting of: halogen, -OH, -CN, -NR _a R _b , optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted C ₃ a -C ₈ cycloalkyl group, and an optionally substituted 3-8 membered heterocyclic group;

R _a and R _b are each independently selected from the group consisting of: H, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, and hydroxy substituted C ₁ -C ₆ alkyl;

m is the number of R ₂ substituents and is 0, 1, 2 or 3;

R ₃ is hydrogen or halogen;

R ₄ is an optionally substituted C ₁ -C ₆ alkyl group;

R _{5 is} selected from the group consisting of hydrogen, an optionally substituted C ₁ -C ₆ alkyl group, an optionally substituted 4-8 membered heterocyclic group, and an optionally substituted C ₃ -C ₈ cycloalkyl group.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof Or prodrug, characterized in that

R _{1 is} selected from the group consisting of: an optionally substituted C ₁ -C ₆ alkyl group, an optionally substituted C ₂ -C ₆ alkenyl group, an optionally substituted C ₂ -C ₆ alkynyl group, an optionally substituted C ₃ -C ₈ a cycloalkyl group, an optionally substituted 4-8 membered heterocyclic group, an optionally substituted C ₁ -C ₆ alkylcarbonyl group, an optionally substituted 4-8 membered heterocyclic carbonyl group, an optionally substituted sulfonyl group, and An optionally substituted sulfinyl group; the optional substituent described in R ₁ is one or more substituents independently selected from the group consisting of halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkane a hydroxy-substituted C ₁ -C ₄ alkyl group, a C ₁ -C ₃ alkoxy group, a -NH(C ₁ -C ₄ alkyl group), a -N(C ₁ -C ₄ alkyl) ₂ group, a C ₃ group C ₈ cycloalkyl and optionally substituted by C ₁ -C ₄ alkyl, halo C ₁ -C ₄ alkyl or hydroxy substituted C ₁ -C ₄ alkyl, containing 1, 2 or 3 selected from nitrogen a 4-8 membered heterocyclic group of oxygen and sulfur;

R _{2 is} selected from the group consisting of: halogen, -OH, -CN, -NR _a R _b , optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted 4- An 8-membered heterocyclic group, and an optionally substituted C ₃ -C ₇ cycloalkyl group; the optional substituents described in R ₂ are one or more substituents independently selected from the group consisting of halogen, -OH, - NH ₂ , -CN, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₄ alkyl), and -N(C ₁ -C ₄ alkyl) ₂ ;

R _a and R _b are each independently selected from the group consisting of: H, C ₁ -C ₄ alkyl, halogenated C ₁ -C ₄ alkyl, and hydroxy-substituted C ₁ -C ₄ alkyl;

m is the number of R ₂ substituents and is 0, 1, 2 or 3;

R ₃ is hydrogen or halogen;

R ₄ is C ₁ -C ₄ alkyl;

R ₅ is selected from: hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₄ -C ₈ heterocyclyl, and optionally substituted C ₃ -C ₈ cycloalkyl group; R ₅ as said optional substituents are one or more substituents independently selected from the group consisting of: _{halo, -OH, -CN, C 1 -C} 4 alkyl, and C ₃ -C ₆ cycloalkyl.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof Or prodrug, characterized in that

R _{1 is} selected from the group consisting of: an optionally substituted C ₁ -C ₄ alkyl group, an optionally substituted C ₃ -C ₆ cycloalkyl group, an optionally substituted 4-6 membered heterocyclic group, an optionally substituted C ₁ -C _{a 4} -alkylcarbonyl group and an optionally substituted 4-6 membered heterocyclylcarbonyl group; the optional substituents described in R ₁ are one or more substituents independently selected from the group consisting of halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkyl, hydroxy substituted C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ - C ₂ alkyl) ₂ , C ₃ -C ₆ cycloalkyl and optionally substituted by C ₁ -C ₂ alkyl, halogenated C ₁ -C ₃ alkyl or hydroxy substituted C ₁ -C ₃ alkyl Containing 1 or 2 4-6 membered heterocyclic groups selected from the group consisting of nitrogen, oxygen and sulfur;

Preferably,

R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, 4-6 membered heterocyclic, C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclic group a hydroxy-substituted C ₁ -C ₃ alkyl-substituted 4-6 membered heterocyclic group, a halogenated C ₁ -C ₃ alkyl-substituted 4-6 membered heterocyclic group, a hydroxy-substituted C ₁ -C ₂ alkane Alkylcarbonyl; R _{1a is} selected from the group consisting of: halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane ₂ , C ₃ -C ₆ cycloalkyl, 4-6 membered heterocyclyl, C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclyl, hydroxy substituted C ₁ -C ₂ alkyl substituted a 4-6 membered heterocyclic group, and a halogenated C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclic group, the heterocyclic group being a heterocyclic group containing 1 or 2 selected from nitrogen and oxygen;

More preferably,

和

R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, hydroxymethyl substituted carbonyl,

with

R _{1a is} selected from the group consisting of: F, -OH, -NH ₂ , -CN, CF ₃ , C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane Base) ₂ ,

R _{1b is} selected from the group consisting of methyl, ethyl and hydroxy substituted propyl groups;

R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate or former thereof Medicament characterized in that R _{2 is} selected from the group consisting of: halogen, -OH, -NH ₂ , -CN, C ₁ -C ₄ alkyl, m is the number of R ₂ substituents, and is 0, 1; preferably, R _{2 is} selected from the group consisting of: F, Cl, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkyl, m is the number of R ₂ substituents, and is 0, 1; more preferably, m is R ₂ substituted The number of bases, and is 0.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof Or prodrug, characterized in that R ₃ is H or F.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof And a prodrug characterized in that R ₄ is a C ₁ -C ₂ alkyl group; preferably, R ₄ is a methyl group.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof Or prodrug, characterized in that

R _{5 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₄ alkyl, and optionally substituted C ₃ -C ₆ cycloalkyl; the optional substituents described in R ₅ are one or two independent a substituent selected from the group consisting of halogen, -OH, -CN, C ₁ -C ₄ alkyl, and C ₃ -C ₅ cycloalkyl;

Preferably,

R _{5 is} selected from:

More preferably, R ₅ is

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate or former thereof Medicine, characterized in that

R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, hydroxymethyl substituted carbonyl,

R _{1a is} selected from the group consisting of: F, -OH, -NH ₂ , -CN, CF ₃ , C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane Base) ₂ ,

R _{1b is} selected from the group consisting of methyl, ethyl and hydroxy substituted propyl groups;

R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl;

m is the number of R ₂ substituents and is 0;

R ₃ is hydrogen or F;

R ₄ is a methyl group;

R ₅ is

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate or former thereof Medicine, characterized in that

R _{1 is} selected from the group consisting of: R _1a (C ₁ -C ₄ alkyl),

R _{1a is} selected from the group consisting of: F, -OH, CF ₃ , C ₁ -C ₃ alkoxy, -NH ₂ , -N(C ₁ -C ₂ alkyl) ₂ ,

R _{1b is} selected from the group consisting of: methyl, and ethyl;

R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl.

In one embodiment, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate thereof, hydrated thereof Or a prodrug, wherein the compound is selected from

In another aspect, the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I, or a stereoisomer, geometric isomer, tautomer, pharmacy thereof An acceptable salt, crystal form, solvate, hydrate or prodrug, and a pharmaceutically acceptable excipient.

In another aspect, the invention provides a compound of Formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form, solvate, hydrate thereof or The use of a prodrug for the preparation of a medicament for preventing, treating, or alleviating a disorder or disease caused by abnormal cell proliferation, autoimmunity, inflammation or infection in a patient.

In one embodiment, the abnormal cell proliferation, autoimmunity, inflammation or infection is caused by a change in a cyclin dependent kinase.

In another aspect, the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystalline form thereof, Solvates, hydrates or prodrugs, as well as pharmaceutically acceptable carriers. In certain embodiments of the pharmaceutical composition, the pharmaceutical composition is formulated for intravenous administration, intramuscular administration, oral administration, rectal administration, inhalation administration, nasal administration, topical administration, ocular administration, or otic administration. In other embodiments of the pharmaceutical composition, the pharmaceutical composition is a tablet, a pill, a capsule, a liquid, an inhalant, a nasal spray solution, a suppository, a solution, an emulsion, an ointment, an eye drop, or an ear drop. Agent. In other embodiments of the pharmaceutical composition, it further comprises one or more additional therapeutic agents.

In another aspect, the invention provides a compound of formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, in the preparation Use in a medicament for preventing, treating, or ameliorating a disorder or disease mediated by one or more abnormal activities of CDK kinase, particularly CDK4/6 kinase.

In another aspect, the invention provides a method of preventing, treating, or ameliorating a disorder or disease mediated by one or more abnormal activities of a CDK kinase, particularly a CDK4/6 kinase, comprising administering to an individual in need of such treatment Administration of an effective amount of a compound of formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, or a pharmaceutical combination comprising the same Things.

In some embodiments of the invention, the disorder or disease includes, but is not limited to, a cell proliferative disorder, an autoimmune disease, an inflammatory disease, or an infectious disease.

In some embodiments of the invention, the cell proliferative disorder includes, but is not limited to, a malignant tumor, such as melanoma, glioblastoma, ovarian cancer, pancreatic cancer, prostate cancer, lung cancer, breast cancer, kidney cancer, cervical cancer. , thyroid cancer, metastasis of secondary sites of primary solid tumors, chronic myeloid leukemia, acute lymphocytic leukemia, other myeloproliferative disorders, papillary thyroid carcinoma, non-small cell lung cancer, and/or mesothelioma .

In some embodiments of the invention, the autoimmune disease includes, but is not limited to, rheumatoid arthritis, systemic lupus erythematosus, idiopathic thrombocytopenic purpura, hemolytic anemia, or psoriasis.

In some embodiments of the invention, the inflammatory disease includes, but is not limited to, osteoarthritis, gouty arthritis, ulcerative colitis, and/or inflammatory bowel disease, and the like.

In some embodiments of the invention, the infectious disease includes, but is not limited to, sepsis, septic shock, endotoxic shock, Gram-negative sepsis, and/or toxic shock syndrome.

Terminology

In the present invention, the terms used in the present invention have the meanings defined below unless explicitly stated otherwise. Terms not expressly defined by the present invention have the general meaning as commonly understood by those skilled in the art.

As used herein, the term "alkyl" refers to a fully saturated straight or branched hydrocarbon group. The alkyl group preferably contains from 1 to 20 carbon atoms, more preferably from 1 to 16 carbon atoms, from 1 to 10 carbon atoms, from 1 to 6 carbon atoms or from 1 to 4 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl Base, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-decyl, n-decyl and the like.

As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon group containing at least one double bond. The alkenyl group preferably contains 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms, 2 to 6 carbon atoms or 2 to 4 carbon atoms. Representative examples of alkenyl include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, pentenyl, isopentenyl, hexenyl, heptenyl, octenyl, and the like.

As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon group containing at least one triple bond. The alkynyl group preferably contains 2 to 20 carbon atoms, more preferably 2 to 10 carbon atoms, 2 to 6 carbon atoms or 2 to 4 carbon atoms. Representative examples of alkynyl include, but are not limited to, ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, pentynyl, isethynyl, hexynyl, heptynyl, octyne Base.

The term "alkoxy" as used herein refers to an alkyl-O- group, wherein alkyl is as defined above. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy, cyclopropyloxy Base, cyclohexyloxy and the like. Preferably, the alkoxy group contains from about 1 to about 6 or from about 1 to about 4 carbons.

As used herein, the term "carbocycle" refers to a saturated or unsaturated monocyclic, bicyclic or tricyclic hydrocarbon group of 3 to 12 carbon atoms. The carbocyclic ring preferably has from 3 to 8 ring carbon atoms, for example containing 3-7, or 4-7 ring carbon atoms. Exemplary monocyclic carbocycles include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cycloheptane, cycloheptene, and the like. Exemplary bicycloalkanes include tetrahydronaphthalene, decahydronaphthalene, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, and the like. Exemplary tricyclic hydrocarbon groups include adamantyl and the like.

As used herein, the term "cycloalkyl" refers to a saturated or unsaturated monocyclic, bicyclic or tricyclic hydrocarbon group of 3 to 12 carbon atoms. The cycloalkyl group preferably contains from 3 to 8 ring carbon atoms, for example, contains 3-7, or 4-7 ring carbon atoms. Exemplary monocyclic hydrocarbon groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, and the like. Exemplary bicyclic hydrocarbon groups include borneol, fluorenyl, hexahydroindenyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, Bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[ 2.2.2] Octyl and so on. Exemplary tricyclic hydrocarbon groups include adamantyl and the like.

As used herein, the term "aryl" is a monocyclic or bicyclic aromatic hydrocarbon group having from 6 to 20 carbon atoms in the ring portion. Preferably, the aryl group is a C ₆ -C ₁₂ aryl group. Non-limiting examples of aryl groups include phenyl, biphenyl, naphthyl or anthryl and the like.

唑基、异

唑基、噻唑基、异噻唑基、三唑基、

二唑基、噻二唑基、四唑基、

三唑基、噻三唑基、吡啶基、嘧啶基、吡嗪基、哒嗪基、

嗪基、二

英基、噻嗪基、三嗪基、5,6-二氢-4H-吡咯并[1,2-b]吡唑基、7H-吡咯并[2,3-d]嘧啶基、咪唑并[1,2-b]哒嗪基、2,3-二氢-1H-咪唑并[1,2-b]吡唑基、咪唑并[2,1-b][1,3]噻唑基、噻吩并[3,2-b]呋喃基、噻吩并[3,2-b]噻吩基、噻吩并[2,3-d][1,3]噻唑基、噻吩并[2,3-d]咪唑基、四唑并[1,5-a]吡啶基、吲哚基、吲嗪基、异吲哚基、苯并呋喃基、异苯并呋喃基、苯并噻吩基、异苯并噻吩基、吲唑基、苯并咪唑基、1,3-苯并

唑基、1,2-苯并异

唑基、2,1-苯并异

唑基、1,3-苯并噻唑基、1,2-苯并异噻唑基、2,1-苯并异噻唑基、苯并三唑基、1,2,3-苯并

二唑基、2,1,3-苯并

二唑基、1,2,3-苯并噻二唑基、2,1,3-苯并噻二唑基、噻吩并吡啶基、嘌呤基、咪唑并[1,2-a]吡啶基、6-氧代-哒嗪-1(6H)-基、2-氧代吡啶-1(2H)-基、6-氧代-哒嗪-1(6H)-基、2-氧代吡啶-1(2H)-基、1,3-苯并二氧杂环戊烯基、喹啉基、异喹啉基、噌啉基、喹唑啉基、喹喔啉基、7-氮杂吲哚基、6-氮杂吲哚基、5-氮杂吲哚基、4-氮杂吲哚基。 As used herein, the term "heteroaryl" refers to a 5-14 membered monocyclic or bicyclic or fused polycyclic aromatic ring containing from 1 to 8 heteroatoms selected from N, O or S. Preferably, the heteroaryl contains 1-3 heteroatoms selected from N, O or S. The heteroaryl group is preferably a 5-10 membered heteroaryl group, more preferably a 5-6 membered heteroaryl group. Preferably, the heteroaryl group includes, but is not limited to, pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl,

Azolyl, different

Azyl, thiazolyl, isothiazolyl, triazolyl,

Diazolyl, thiadiazolyl, tetrazolyl,

Triazolyl, thiatriazole, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl,

Azinyl, two

English, thiazinyl, triazinyl, 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazolyl, 7H-pyrrolo[2,3-d]pyrimidinyl, imidazo[1 ,2-b]pyridazinyl, 2,3-dihydro-1H-imidazo[1,2-b]pyrazolyl, imidazo[2,1-b][1,3]thiazolyl, thieno [3,2-b]furanyl, thieno[3,2-b]thienyl, thieno[2,3-d][1,3]thiazolyl, thieno[2,3-d]imidazolyl , tetrazolo[1,5-a]pyridinyl, fluorenyl, pyridazinyl, isodecyl, benzofuranyl, isobenzofuranyl, benzothienyl, isobenzothienyl, anthracene Azolyl, benzimidazolyl, 1,3-benzo

Azolyl, 1,2-benziso

Azolyl, 2,1-benziso

Azyl, 1,3-benzothiazolyl, 1,2-benzisothiazolyl, 2,1-benzisothiazolyl, benzotriazolyl, 1,2,3-benzo

Diazolyl, 2,1,3-benzoene

Diazolyl, 1,2,3-benzothiadiazolyl, 2,1,3-benzothiadiazolyl, thienopyridinyl, fluorenyl, imidazo[1,2-a]pyridyl, 6-oxo-pyridazine-1(6H)-yl, 2-oxopyridine-1(2H)-yl, 6-oxo-pyridazine-1(6H)-yl, 2-oxopyridine-1 (2H)-yl, 1,3-benzodioxolyl, quinolyl, isoquinolinyl, porphyrinyl, quinazolinyl, quinoxalinyl, 7-azaindole , 6-azaindolyl, 5-azaindenyl, 4-azaindenyl.

As used herein, the term "heterocycle" refers to a fully saturated or unsaturated, aromatic or non-aromatic cyclic group, for example, which is a 4- to 7-membered monocyclic, 7- to 12-membered bicyclic ring or A 10- to 15-membered tricyclic ring system containing at least one hetero atom on a ring containing at least one carbon atom. Each ring of the hetero atom containing a hetero atom may have 1 to 6, preferably 1, 2 or 3 hetero atoms selected from a nitrogen atom, an oxygen atom or a sulfur atom, wherein the nitrogen and sulfur hetero atoms may also be Optionally oxidized. Preferably, the heterocyclic ring is a 4- to 7-membered monocyclic heterocyclic ring.

唑、

唑烷、异

唑啉、异

唑、噻唑、噻二唑、噻唑烷、异噻唑、异噻唑烷、呋喃、四氢呋喃、噻吩、

二唑、哌啶、哌嗪、2-氧代哌嗪、2-氧代哌啶、2-氧代吡咯烷、2-氧代氮杂

、氮杂

、4-哌啶酮、吡啶、吡嗪、嘧啶、哒嗪、四氢吡喃、吗啉、硫吗啉、硫吗啉亚砜、硫吗啉砜、1,3-二氧戊环和四氢-1,1-二氧代噻吩、1,1,4-三氧代-1,2,5-噻二唑烷-2-等。 Exemplary monocyclic heterocycles include pyrrolidine, pyrrole, pyrazole, oxetane, pyrazoline, imidazole, imidazoline, imidazolidine, triazole,

Azole,

Oxazolidine

Oxazoline

Oxazole, thiazole, thiadiazole, thiazolidine, isothiazole, isothiazolidine, furan, tetrahydrofuran, thiophene,

Diazole, piperidine, piperazine, 2-oxopiperazine, 2-oxopiperidine, 2-oxopyrrolidine, 2-oxoaza

Aza

, 4-piperidone, pyridine, pyrazine, pyrimidine, pyridazine, tetrahydropyran, morpholine, thiomorpholine, thiomorpholine sulfoxide, thiomorpholine sulfone, 1,3-dioxolane and tetra Hydrogen-1,1-dioxothiophene, 1,1,4-trioxo-1,2,5-thiadiazolidine-2- and the like.

嗪、苯并

唑、苯并噻吩、苯并噻嗪、奎宁环、喹啉、四氢喹啉、十氢喹啉、异喹啉、四氢异喹啉、十氢异喹啉、苯并咪唑、苯并吡喃、吲嗪、苯并呋喃、色原酮、香豆素、苯并吡喃、噌啉、喹喔啉、吲唑、吡咯并吡啶、呋喃并吡啶(例如呋喃并[2,3-c]吡啶、呋喃并[3,2-b]-吡啶]或呋喃并[2,3-b]吡啶)、二氢异吲哚、1,3-二氧代-1,3-二氢异吲哚-2-、二氢喹唑啉(例如3,4-二氢-4-氧代-喹唑啉)、酞嗪等。 Exemplary bicyclic heterocycles include anthracene, indoline, benzothiazole, benzo

Oxazine, benzo

Azole, benzothiophene, benzothiazine, quinuclidine, quinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, tetrahydroisoquinoline, decahydroisoquinoline, benzimidazole, benzo Pyran, pyridazine, benzofuran, chromone, coumarin, benzopyran, porphyrin, quinoxaline, oxazole, pyrrolopyridine, furopyridine (eg furo[2,3-c] Pyridine, furo[3,2-b]-pyridine] or furo[2,3-b]pyridine), dihydroisoindole, 1,3-dioxo-1,3-dihydroisoindole Indole-2-, dihydroquinazoline (e.g., 3,4-dihydro-4-oxo-quinazoline), pyridazine, and the like.

、二噻吩并氮杂

、苯并吲哚、菲咯啉、吖啶、菲啶、吩

嗪、吩噻嗪、呫吨、咔啉等。 Exemplary tricyclic heterocycles include carbazole, dibenzoazepine

Dithienoazepine

Benzopyrene, phenanthroline, acridine, phenanthridine, phenanthrene

Oxazine, phenothiazine, xanthene, porphyrin and the like.

"Heterocyclyl" or "heterocycloalkyl" refers to a group formed by the loss of one or more hydrogen atoms from a heterocycle as defined above. The heterocyclic group can be attached at a hetero atom or a carbon atom. Preferably, the heterocycloalkyl group is a 4-8 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur.

The term "Boc" means a tert-butoxycarbonyl group.

As used herein, the term "halogen" or "halo" refers to fluoro, chloro, bromo and iodo. Herein, a group such as an alkyl group, an alkenyl group, an alkoxy group, a carbocyclic ring, a cycloalkyl group, a heteroaryl group, a heterocyclic group, a heterocyclic group, a carbonyl group, a sulfonyl group, a sulfinyl group or the like may be substituted with a substituent. The substituents include, but are not limited to, OH, Boc, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclic ring. a group; NRR', C(O)R, C(O)NRR' or C(O)OR, and each of R and b' is independently selected from the group consisting of: H and a substituted or unsubstituted alkyl group.

The term "optional," "optional," or "optionally" as used herein means that the substitution pattern, event, or circumstance described subsequently may or may not occur, and that the description includes the occurrence of the substitution pattern and The case where the substitution mode does not occur. For example, "optionally substituted alkyl" includes "unsubstituted alkyl" and "substituted alkyl" as defined herein. It will be understood by those skilled in the art that for any group containing one or more substituents, the group does not include any spatially impractical, chemically incorrect, synthetically infeasible And/or an inherently unstable substitution pattern.

The term "substituted" or "substituted by" as used herein means that one or more hydrogen atoms on a given atom or group are replaced by one or more substituents selected from a given group of substituents, conditions. It does not exceed the normal valence of the given atom. When the substituent is oxo (ie, =0), then two hydrogen atoms on a single atom are replaced by oxygen. Such combinations are permissible only if the combination of substituents and/or variables results in a chemically correct and stable compound. A chemically correct and stable compound means that the compound is sufficiently stable to be separated from the reaction mixture and to determine the chemical structure of the compound, and can then be formulated into at least a practically useful formulation. For example, the term "substituted" or "substituted by" as used herein, unless the substituent is specifically recited, means that one or more hydrogen atoms on a given atom or group are independently or more, e.g., three or four substituents, the substituents are independently selected from: deuterium (D), halogen, -OH, a mercapto group, a cyano group, -CD _3, -C ₁ -C ₆ alkyl (preferably -C _1-3 alkyl), C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, cycloalkyl (preferably 3-8 membered cycloalkyl), aryl, heterocyclic (preferably a 3-8 membered heterocyclic group), a heteroaryl group, an aryl-C ₁ -C ₆ alkyl group, a heteroaryl-C ₁ -C ₆ alkyl group, a C ₁ -C ₆ haloalkyl group, and OC ₁ -C ₆ alkyl (preferably -OC ₁ -C ₃ alkyl), -OC ₂ -C ₆ alkenyl, -OC ₁ -C ₆ alkylphenyl, -C ₁ -C ₆ alkyl-OH ( Preferred is -C ₁ -C ₄ alkyl-OH), -C ₁ -C ₆ alkyl-SH, -C ₁ -C ₆ alkyl-OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ haloalkyl, -NH ₂ , -C ₁ -C ₆ alkyl-NH ₂ (preferably -C ₁ -C ₃ alkyl-NH ₂ ), -N(C ₁ -C ₆ alkyl) ₂ (preferably -N(C ₁ - C ₃ alkyl) ₂ ), -NH(C ₁ -C ₆ alkyl) (preferably -NH(C ₁ -C ₃ alkyl)), -N(C ₁ -C ₆ alkyl) (C ₁ -C ₆ alkylphenyl), -NH (C ₁ -C ₆ alkylphenyl), nitro, -C(O)-OH, -C(O)OC ₁ -C ₆ alkyl (preferably -C(O)OC ₁ -C ₃ alkyl), -CONRiRii (wherein Ri and Ri are H, D and C _1-6 alkyl, preferably C _1-3 alkyl), -NHC(O)(C ₁ -C ₆ alkyl), -NHC(O)(phenyl) , -N(C ₁ -C ₆ alkyl)C(O)(C ₁ -C ₆ alkyl), -N(C ₁ -C ₆ alkyl)C(O)(phenyl), -C(O C ₁ -C ₆ alkyl, -C(O)heteroaryl (preferably -C(O)-5-7 membered heteroaryl), -C(O)C ₁ -C ₆ alkylphenyl, - C(O)C ₁ -C ₆ haloalkyl, -OC(O)C ₁ -C ₆ alkyl (preferably -OC(O)C ₁ -C ₃ alkyl), -S(O) ₂ -C ₁ - C ₆ alkyl, -S(O)-C ₁ -C ₆ alkyl, -S(O) ₂ -phenyl, -S(O) ₂ -C ₁ -C ₆ haloalkyl, -S(O) ₂ NH ₂ , -S(O) ₂ NH(C ₁ -C ₆ alkyl), -S(O) ₂ NH(phenyl), -NHS(O) ₂ (C ₁ -C ₆ alkyl), -NHS (O) ₂ (phenyl) and -NHS(O) ₂ (C ₁ -C ₆ haloalkyl), wherein the alkyl group, the cycloalkyl group, the phenyl group, the aryl group, the heterocyclic group and the heteroaryl group are Each of which is optionally further substituted with one or more substituents selected from the group consisting of halogen, -OH, -NH ₂ , cycloalkyl, 3-8 membered heterocyclyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ haloalkyl-, -OC ₁ -C ₄ alkyl, -C ₁ -C ₄ alkyl-OH, -C ₁ -C ₄ alkyl-OC ₁ -C ₄ alkyl, -OC ₁ -C ₄ haloalkyl, cyano, nitro, -C(O)-OH, -C(O)OC ₁ -C ₆ alkyl, -CON(C ₁ -C ₆ alkyl) ₂ , -CONH(C ₁ -C ₆ alkyl), -CONH ₂ , -NHC(O)(C ₁ -C ₆ alkyl), -NH(C ₁ - C ₆ alkyl)C(O)(C ₁ -C ₆ alkyl), -SO ₂ (C ₁ -C ₆ alkyl), -SO ₂ (phenyl), -SO ₂ (C ₁ -C ₆ haloalkane) Base), -SO ₂ NH ₂ , -SO ₂ NH(C ₁ -C ₆ alkyl), -SO ₂ NH(phenyl), -NHSO ₂ (C ₁ -C ₆ alkyl), -NHSO ₂ (benzene Base) and -NHSO ₂ (C ₁ -C ₆ haloalkyl). When one atom or group is substituted with a plurality of substituents, the substituents may be the same or different.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effects and properties of the compounds of the invention, and which are not biologically or otherwise undesirable. Non-limiting examples of such salts include non-toxic, inorganic or organic base or acid addition salts of the compounds of the invention. In many cases, the compounds of the invention are capable of forming acid and/or base salts due to the presence of amino and/or carboxyl groups or groups similar thereto. A pharmaceutically acceptable acid addition salt can be formed from a mineral acid and an organic acid. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, Mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; ammonium, potassium, sodium, calcium, and magnesium salts are particularly preferred. Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins and the like, especially such as isopropylamine, Trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound (basic or acidic moiety) by conventional chemical methods. In general, the salts can be prepared by reacting the free acid form of the compound with a stoichiometric amount of a suitable base (eg, a hydroxide, carbonate, bicarbonate, etc. of Na, Ca, Mg or K). The reaction or reaction of the free base form of the compound with a stoichiometric amount of the appropriate acid. Such a reaction is usually carried out in water or an organic solvent or a mixed solvent of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred where practicable. Other suitable salts can be found in Remington's Pharmaceutical Sciences, 20th Edition, Mack Publishing Company, Easton, Pa., (1985), which is incorporated herein by reference.

The term "pharmaceutically acceptable excipient" as used herein includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (eg, antibacterial, antifungal), isotonic agents. , absorption delaying agents, salts, preservatives, pharmaceuticals, pharmaceutical stabilizers, binders, excipients, disintegrating agents, lubricants, sweeteners, flavoring agents, dyes, similar substances, and combinations thereof, It is well known to those of ordinary skill in the art (see, for example, Remington's Pharmaceutical Sciences, 18th ed., Mack Printing Company, 1990, pp. 1289-1329, incorporated herein by reference. ). Unless any conventional carrier is incompatible with the active ingredient, it can be considered for use in a therapeutic or pharmaceutical composition.

As used herein, the term "solvate" means a solvent addition form comprising a stoichiometric or non-stoichiometric solvent. If the solvent is water, the solvate formed is a hydrate, and when the solvent is ethanol, the solvate formed is an ethanolate. Hydrate is formed by one or more molecules of water and one molecule of said substance, wherein water retains its molecular state of H ₂ O, such a combination can form one or more hydrates, such as hemihydrate, Hydrates and dihydrates.

As used herein, "prodrug" refers to a chemically modified active or inactive compound which, upon administration to an individual, becomes a compound of the invention by physiological action (eg, hydrolysis, neogeneration, etc.) in the body. Adaptability and techniques for making and using prodrugs are well known to those skilled in the art.

The term "therapeutically effective amount" of a compound of the invention refers to an amount of a compound of the invention which can elicit an individual's biological or medical response or ameliorate symptoms, slow or delay the progression of the disease, or prevent disease, and the like.

As used herein, the term "individual" refers to an animal. Preferably, the animal is a mammal. Individuals also refer to, for example, primates (eg, humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, and the like. In a preferred embodiment, the individual is a human.

As used herein, the term "inhibiting" refers to the alleviation or inhibition of a particular patient, condition or disorder or disease, or a significant decrease in biological activity or process baseline activity.

As used herein, in one embodiment the term "treating" any disease or condition refers to ameliorating a disease or condition (ie, preventing or slowing the progression of a disease or at least one of its clinical symptoms). In another embodiment, "treating" refers to ameliorating at least one physical parameter that may not be perceived by a patient. In another embodiment, "treating" refers to modulating a disease or condition either physically (eg, to stabilize a detectable condition) or physiologically (eg, to stabilize a parameter of the body) or both.

Any asymmetric carbon atom in the compounds of the invention may exist in the (R)-, (S)- or (R,S)-configuration, preferably in the (R)- or (S)-configuration. Substituents for unsaturated bond atoms, if possible, may exist in cis-(Z)- or trans-(E)- form. Thus, the compounds of the invention may exist as one of the possible isomers or as a mixture thereof, for example, as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (Enantiomer), racemate or a mixture thereof.

Any resulting mixture of isomers can be separated into pure geometric or optical isomers, diastereomers, racemates, for example by chromatography and/or fractional crystallization, depending on the physicochemical differences of the components. .

The main advantage of the present invention is that the compounds of the present invention have high selectivity for CDK4/6 kinase, have fewer side effects, and have high resistance to drug resistance (i.e., have high activity against diseases that have been resistant to current kinase inhibitors). It has high bioavailability and high clinical application value, and it has good pharmacokinetic properties, has a good blood-brain distribution, and can cross the blood-brain barrier to inhibit the growth of brain tumors.

Detailed ways

In the present application, when the chemical name and the structural formula are inconsistent, the structural formula shall prevail, unless the chemical name may be inferred from the context instead of the structural formula.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight and parts by weight.

The experimental materials and reagents used in the following examples are available from commercially available sources unless otherwise specified.

In each of the examples, the experimental instrument description (for example, ¹ H NMR is recorded by Varian Mercury-300 or Varian Mercury-400 type nuclear magnetic resonance apparatus, and ¹³ C NMR is performed by Varian Mercury-400 or Varian Mercury-500 or Varian Mercury-600 type nuclear magnetic Resonance recording, chemical shift is expressed in δ (ppm); mass spectrometry is recorded by Finnigan/MAT-95 (EI) and Finnigan LCQ/DECA and Micromass Ultra Q-TOF (ESI) mass spectrometer; 200-300 mesh).

Among them, the Chinese name of the reagent represented by the chemical formula or the English alphabet abbreviation is as follows:

iPrOH: isopropanol; EtOH: ethanol; DCM: dichloromethane; TFA: trifluoroacetic acid; MeOH: methanol; NaOH: sodium hydroxide; HCl: hydrogen chloride; TEA: triethylamine; Raney Ni: Raney nickel; , 4-dioxane: 1,4-dioxane; NaH: sodium hydride; H ₂ O: water; Pd/C: palladium/carbon; H ₂ : hydrogen; HATU: 2-(7-oxidized benzotriazine Azole)-N,N,N',N'-tetramethyluron hexafluorophosphate; DMF: N,N-dimethylformamide; THF: tetrahydrofuran; Boc ₂ O: di-tert-butyl dicarbonate; NBS : N-bromosuccinimide; NCS: N-chlorosuccinimide; NIS: N-iodosuccinimide; MeCN: acetonitrile; DIPEA: N, N-diisopropyl B Amine; NaBH ₄ : sodium borohydride; AcOH: acetic acid; ethyl acetate: ethyl acetate; NaBH ₃ CN: sodium cyanoborohydride; K ₂ CO ₃ : potassium carbonate; Cs ₂ CO ₃ : cesium carbonate; nBuLi: n-butyl Lithium lithium; LiAlH ₄ : lithium aluminum hydride; Pd(dppf)Cl ₂ : [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; KOAc: potassium acetate. Fumaronitrile: fumaric acid nitrile; P(nBu) ₃ : tri-n-butylphosphine; LDA: lithium diisopropylamide; LiOH: lithium hydroxide; MeI: methyl iodide; EtI: ethyl iodide; (CH ₂ O) _n : paraformaldehyde; HCO ₂ H: formic acid; CH ₃ COCl: acetyl chloride; HBTU: O-benzotriazole-tetramethylurea hexafluorophosphate; Pd ₂ (dba) ₃ : tris (dibenzylidene) Acetone) dipalladium; Xantphos: 4,5-bisdiphenylphosphino-9,9-dimethyloxaxan; DCE: dichloroethane; TLC: thin layer chromatography; DIEA: N, N-II Isopropylethylamine; RT: retention time; LCMS: liquid chromatography-mass spectrometry; HEPES: 4-hydroxyethylpiperazineethanesulfonic acid; EGTA: ethylene glycol bis(2-aminoethyl ether) tetraacetic acid; DTT: dithiothreitol; EDTA: ethylenediaminetetraacetic acid.

Synthesis of key intermediates

Intermediate 1:

First step: 2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7, 8-Dihydro-1,6-naphthyridin-6(5H)-carboxylic acid tert-butyl ester

Add 5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine (270 mg, 0.89 mmol) to a dry single-mouth flask and 2-Chloro-7,8-dihydro-1,6-naphthyridin-6(5H)-carboxylic acid tert-butyl ester (360.2 mg, 1.34 mmol), potassium t-butoxide (200 mg, 1.78 mmol), Pd ( OAc) ₂ (99 mg, 0.45 mmol), then isopropanol (10 mL) was added, and the mixture was heated to 90 ° C under nitrogen atmosphere and stirred for 5 hours. The reaction was completed by LCMS. The reaction was cooled to room temperature, filtered and filtered. The cake was washed with a small amount of isopropyl alcohol, washed with a small amount of water and dried under reduced pressure to give the title compound 2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-carbazole)- 5-Benzylpyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-carboxylic acid tert-butyl ester (300 mg, yellow solid). LCMS: 536.4 (M+H) ⁺ .

Second step: N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7 ,8-tetrahydro-1,6-naphthyridin-2-amine

The compound 2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7,8- Dihydro-1,6-naphthyridin-6(5H)-carboxylic acid tert-butyl ester (300 mg, 0.56 mmol) was added to a single-necked flask, followed by methanol 15 (mL), 4N hydrochloric acid dioxane (5 mL) The reaction mixture was stirred at room temperature for 2 hr. EtOAc (EtOAc m. -(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro -1,6-naphthyridin-2-amine hydrochloride (210 mg, pale yellow solid). LCMS: 436.1 (M+H) ⁺ . ¹ H NMR (400 MHz, DMSO) δ 11.25 (s, 1H), 9.95 (s, 2H), 8.82 - 8.81 (d, 1H, J = 4 Hz), 8.53 (s, 1H), 8.27-8.25 (d, 1H) , J=4Hz), 8.04(s,1H), 7.73-7.70(d,1H,J=12Hz), 4.37(s,2H), 4.19(s,2H),3.64-3.61(m,1H), 3.49 (m, 2H), 3.23 (s, 3H), 1.51-1.49 (d, 6H, J = 8 Hz).

Intermediate 2 was prepared according to the synthesis method of Intermediate 1, using the corresponding starting materials.

General synthetic method for target compounds: Reductive amination

The above intermediate amines (1 eq.), the corresponding aldehydes or ketones (1-3 eq.), suitable solvents such as DCE, DMF, THF, etc. (suitable amount), acetic acid (0.1) are added sequentially to the dried reaction flask at room temperature. -1.5 eq.), a reducing agent such as NaBH(OAc) ₃ , NaBH ₃ CN or the like (0.5-5 eq.), reacted at room temperature or heated until the end of the reaction. After completion of the reaction, it was quenched by the addition of saturated sodium hydrogencarbonate and extracted twice with dichloromethane or ethyl acetate. The organic phase was combined, dried and filtered, evaporated, evaporated, evaporated, evaporated,

General synthetic method of target compound II: substitution reaction

The above intermediate amines (1 eq.) are sequentially added to a dry round bottom flask at room temperature, a suitable solvent such as DMF or the like (appropriate amount), a corresponding halogenated product, an epoxy compound or a sulfonic acid ester (1.2 eq. A suitable base such as triethylamine, potassium carbonate, sodium carbonate or the like (1-2 eq.) is allowed to react at room temperature or for 12 hours. Water is added to the reaction mixture, and the mixture is extracted three times with an organic solvent such as dichloromethane or ethyl acetate. The organic phase was combined, dried and filtered, evaporated, evaporated, evaporated, evaporated,

General synthetic method for target compounds: condensation reaction

Each of the above intermediate amines (1 eq.), a suitable solvent such as DMF or the like (suitable amount), a condensing agent such as HBTU, HATU (1-2 eq), the corresponding carboxylic acid or activated are sequentially added to a dry round bottom flask at room temperature. the acid material (e.g., carboxylic anhydride) (1.2eq.), a suitable organic base such as DIPEA, et ₃ N and the like (1-5 eq.), at room temperature for 12 hours. Water is added to the reaction mixture, and the mixture is extracted three times with an organic solvent such as dichloromethane or ethyl acetate. The organic phase was combined, dried and filtered, evaporated, evaporated, evaporated, evaporated, or

The above intermediate amines (1 eq.) are added sequentially in a dry round bottom flask at room temperature, a suitable solvent such as DCM (yield), the corresponding acid chloride starting material (1.2 eq.), a suitable organic base such as DIPEA, Et ₃ N (1-5 eq.), reacted at room temperature for 12 hours. Water is added to the reaction mixture, and the mixture is extracted three times with an organic solvent such as dichloromethane or ethyl acetate. The organic phase was combined, dried and filtered, evaporated, evaporated, evaporated, evaporated,

General synthetic method for target compounds 4: Addition reaction

The above intermediate amines (1 eq.) are sequentially added to a dry round bottom flask at room temperature, a suitable solvent such as MeOH, DMF, etc. (suitable amount), corresponding olefin starting material (1.2 eq.), a suitable base such as triethyl Amine, potassium carbonate, sodium carbonate or the like or CuBr (1-2 eq.), and reacted at room temperature or for 12 hours. Water is added to the reaction mixture, and the mixture is extracted three times with an organic solvent such as dichloromethane or ethyl acetate. The organic phase was combined, dried and filtered, evaporated, evaporated, evaporated,

Example 1: (6-(2-(Dimethylamino)ethyl)-N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl) Pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

Add (N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7,8 to a 100 mL flask -tetrahydro-1,6-naphthyridin-2-amine) (300 mg, 0.72 mmol) and 2-bromo-N,N-dimethylethylamine (164 mg, 1.08 mmol) in 10 mL of DMF, then DIEA (372 mg, 2.88 mmol) was added. The reaction was stirred at 80 ° C overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted twice with dichloromethane (2×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate Chromatographic separation of the product (6-(2-(dimethylamino)ethyl)-N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl) Pyrimidine-2-yl)-5,6,7,8 tetrahydro-1,6-naphthyridin-2-amine) 24 mg, yellow powder, yield 6.8%.

¹ H NMR (400 MHz, CDCl 3 ) δ 8.69 (s, 1H), δ 8.38 (d, 1H, J = 2.8), δ 8.24 (d, 1H, J = 8.0), δ 8.01 - 8.04 (m, 2H), δ 7.72 (d, 1H, J = 8.8), δ 7.35 (d, 1H, J = 8.4), δ 4.17 (s, 3H), δ 3.65 (s, 2H), δ 3.48 -3.51 (m, 1H), δ 2.97 (s, 2H), δ 2.87-2.89 (m, 2H), δ 2.67-2.71 (m, 2H), δ 2.55-2.56 (m, 2H), δ 2.29 (s, 6H), δ 1.58 (d, 6H, J = 6.8). LCMS: 489.3 (M+H); RT = 1.13 min

Example 2: (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2 -(pyrrolidin-1-yl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine)

Add (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6 to a 100 mL flask ,7,8-tetrahydro-1,6-naphthyridin-2-amine) (90 mg, 0.21 mmol) and 1-(2-chloroethyl)pyrrolidine (42 mg, 0.31 mmol) in 10 mL of DMF Then, DIEA (108 mg, 0.84 mmol) was added. The reaction was stirred at 80 ° C overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted with dichloromethane (3×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate Chromatography product (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2 -(Pyrrolidin-1-yl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine) 15 mg, yellow powder, yield 13.42%.

¹ H NMR (400 MHz, CDCl 3 ) δ 8.50 (s, 1H), δ 8.39 (d, 1H, J = 3.6), δ 8.19 (d, 1H, J = 8.4), δ 7.99 (s, 1H) , δ 7.75 (d, 1H, J = 12.8), δ 7.36 (d, 1H, J = 8.8), δ 4.20 (s, 3H), δ 3.66 (s, 2H), δ 3.47-3.50 (m, 1H), δ 2.95-2.98 (m, 2H), δ 2.87-2.90 (m, 2H), δ 2.75 (s, 4H), δ 2.59 (s, 4H), δ 1.80- 1.84 (m, 4H), δ 1.58 (d, 6H, J = 7.2). LCMS: 533.3 (M+H); RT = 1.10 min

Example 3: (6-(2-(Diethylamino)ethyl)-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-carbazole- 5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine)

Add (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6 to a 100 mL flask ,7,8-tetrahydro-1,6-naphthyridin-2-amine) (100 mg, 0.23 mmol) and 2-chloro-N,N-diethylethylamine (47 mg, 0.35 mmol) in 10 mL In DMF, DIEA (89 mg, 0.69 mmol) was then added. The reaction was stirred at 80 ° C overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted twice with dichloromethane (2×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate Chromatographic separation of the product (6-(2-(diethylamino)ethyl)-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-carbazole- 5-Methyl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine) 30 mg, yellow powder, yield 24.43%.

¹ H NMR (400 MHz, CDCl 3 ) δ 8.50 (s, 1H), δ 8.38 (d, 1H, J = 3.6), δ 8.19 (d, 1H, J = 8.4), δ 7.91 (s, 1H) , δ 7.75 (d, 1H, J = 12.8), δ 7.37 (d, 1H, J = 8.4), δ 4.20 (s, 3H), δ 3.65 (s, 2H), δ 3.45-3.52 (m, 1H), δ 2.95-2.98 (m, 2H), δ 2.86-2.89 (m, 2H), δ 2.67-2.73 (m, 4H), δ 2.57-2.62 (m, 4H), δ 1.58 (d, 6H, J = 6.8), δ 1.04-1.08 (t, 6H, J = 6.8). LCMS: 535.3 (M+H); RT = 1.19 min

Example 4: (N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(3 -(pyrrolidin-1-yl)propyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine)

Add (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6 to a 100 mL flask ,7,8-tetrahydro-1,6-naphthyridin-2-amine) (90 mg, 0.21 mmol) and N-(3-chloropropyl)tetrahydropyrrolidine (46 mg, 0.31 mmol) in 10 mL In DMF, DIEA (82 mg, 0.63 mmol) was then added. The reaction was stirred at 80 ° C overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted with dichloromethane (3×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate Chromatographic separation of the product (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(3 -(Pyrrolidin-1-yl)propyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine) 18 mg, yellow solid, yield 15.93%.

¹ H NMR (400 MHz, CDCl 3 ) δ 8.51 (s, 1H), δ 8.38 (d, 1H, J = 4.0), δ 8.19 (d, 1H, J = 8.8), δ 7.97 (s, 1H) , δ 7.73 (d, 1H, J = 12.8), δ 7.39 (d, 1H, J = 8.4), δ 4.20 (s, 3H), δ 3.62 (s, 2H), δ 3.47-3.51 (m, 1H), δ 3.28 (s, 4H), δ 3.14 - 3.19 (m, 2H), δ 2.92 - 2.95 (m, 2H), δ 2.82 - 2.85 (m, 2H), δ 2. 66-2.69 (m, 2H), δ 2.18-2.22 (m, 2H), δ 2.14 (s, 4H), δ 1.58 (d, 6H, J = 7.2). LCMS: 547.3 (M+H) ;RT=1.18min

The compound of Example 5-13 was prepared in the same manner as in Example 1.

Example 5: N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2-fluoroethyl) -5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl3-d1) δ 8.71 (s, 1H), 8.38 (d, 1H, J = 4.0 Hz), 8.27 (d, 1H, J = 8.4 Hz), 8.05 (d, 1H, J = 9.6 Hz), 7.92 (s, 1H), 7.73 (d, 1H, J = 9.2 Hz), 7.38 (d, 1H, J = 8.4 Hz), 4.76 (t, 1H, J = 4.8 Hz), 4.64 (t , 1H, J = 4.8 Hz), 4.19 (s, 3H), 3.74 (s, 2H), 3.47-3.52 (m, 1H), 2.89-3.00 (m, 6H), 1.60 (d, 6H, J = 6.8 Hz). LCMS: 464.3 (M+H)+; RT = 1.137 min.

Example 6: N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2- Fluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl3-d1) δ 8.52 (d, 1H, J = 1.2 Hz), 8.39 (d, 1H, J = 4.0 Hz), 8.22 (d, 1H, J = 8.4 Hz), 7.91 (s) , 1H), 7.76 (d, 1H, J = 12.8 Hz), 7.39 (d, 1H, J = 8.4 Hz), 4.76 (t, 1H, J = 4.8 Hz), 4.64 (t, 1H, J = 5.0 Hz) ), 4.22 (s, 3H), 3.75 (s, 2H), 3.48-3.52 (m, 1H), 2.89-3.00 (m, 6H), 1.60 (d, 6H, J = 7.2 Hz). LCMS: 482.3 ( M+H)+; RT=1.397min.

Example 7: N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2,2,2- Trifluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl3-d1) δ 8.71 (s, 1H), 8.41 (d, 1H, J = 4.0 Hz), 8.28 (d, 1H, J = 8.4 Hz), 8.11 (s, 1H), 8.05 (d, 1H, J = 9.2 Hz), 7.73 (d, 1H, J = 9.2 Hz), 7.36 (d, 1H, J = 8.4 Hz), 4.17 (s, 3H), 3.88 (s, 2H), 3.48 -3.52 (m, 1H), 3.18-3.25 (q, 2H), 3.09-3.12 (m, 2H), 2.98-3.00 (m, 2H), 1.61 (d, 6H, J = 7.2 Hz). LCMS: 500.2 (M+H)+; RT=1.09min.

Example 8: N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2, 2,2-trifluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl3-d1) δ 8.51 (d, 1H, J = 1.2 Hz), 8.43 (d, 1H, J = 3.6 Hz), 8.23 (d, 1H, J = 8.4 Hz), 8.15 (s) , 1H), 7.75 (d, 1H, J = 12.8 Hz), 7.38 (d, 1H, J = 8.8 Hz), 4.21 (s, 3H), 3.88 (s, 2H), 3.47-3.53 (m, 1H) , 3.18-3.25(q, 2H), 3.09-3.12 (m, 2H), 2.98-3.01 (m, 2H), 1.59 (d, 6H, J = 6.8 Hz). LCMS: 518.2 (M+H)+; RT=1.18min.

Example 9: 3-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) -7,8-Dihydro-1,6-naphthyridin-6(5H)-yl)-2,2-dimethylpropan-1-ol

¹ H NMR (400 MHz, CDCl3-d1) δ 8.50 (s, 1H), 8.38 (d, 1H, J = 4.0 Hz), 8.20 (d, 1H, J = 8.4 Hz), 7.92 (s, 1H), 7.82 (d, 1H, J = 12.8 Hz), 7.35 (d, 1H, J = 8.8 Hz), 5.49 (s, 1H), 4.20 (s, 3H), 3.76 (s, 2H), 3.55 (s, 2H) , 3.48-3.53 (m, 1H), 2.97 (s, 4H), 2.60 (s, 2H), 1.58 (d, 6H, J = 6.8 Hz), 1.00 (s, 6H). LCMS: 522.3 (M+H )+;RT=1.764min.

Example 10: N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2-methoxy B -5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CD ₃ OD) δ 8.72 (s, 1H), 8.45-8.44 (d, 1H, J = 4 Hz), 8.20-8.18 (d, 1H, J = 8 Hz), 8.04-8.02 (d, 1H, J=8Hz), 7.62-7.60 (d, 1H, J=8Hz), 7.45-7.43 (d, 1H, J=8Hz), 4.16(s, 3H), 3.67-3.61(m, 5H), 3.37 (s, 3H), 2.92 (s, 4H), 2.79-2.76 (t, 2H, J = 12 Hz), 1.59-1.57 (d, 6H, J = 8 Hz). LCMS: 476.3 (M + H) ⁺ ; RT = 1.94 min.

Example 11: (N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2 -methoxyethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine)

¹ H NMR (400 MHz, DMSO) δ 10.25 (s, 1H), 8.67 (d, 1H, J = 4.0 Hz), 8.53 (s, 1H), 8.14 (d, 1H, J = 8.8 Hz), 7.65-7.72 (m, 2H), 4.18 (s, 2H), 4.10 (s, 3H), 3.78-4.10 (m, 2H), 3.75-3.78 (t, 2H, J = 4.4 Hz), 3.59-3.62 (t, 1H) , J = 6.8 Hz), 3.48 (s, 2H), 3.36 (s, 3H), 3.12 (s, 2H), 1.50 (d, 6H, J = 7.2 Hz). LCMS: 494.3 (M+H) ⁺ ; RT=1.26min

Example 12: 2-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) -7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanol

¹ H NMR (400 MHz, CDCl3-d1) δ 8.50 (d, 1H, J = 0.80 Hz), 8.38 (d, 1H, J = 6.0 Hz), 8.23 (d, 1H, J = 8.4 Hz), 7.91 (s) , 1H), 7.75 (d, 1H, J = 12.8 Hz), 7.39 (d, 1H, J = 8.4 Hz), 4.21 (s, 3H), 3.77-3.80 (m, 4H), 3.47-3.51 (m, 1H), 3.00 (s, 4H), 2.83 (t, 2H, J = 5.2 Hz), 1.58 (d, 6H, J = 7.2 Hz). LCMS: 480.3 (M + H) +; RT = 1.280 min.

Example 13: N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(2, 2-difluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.51 (s, 1H), 8.45-8.30 (m, 1H), 8.29-8.20 (m, 1H), 7.80-7.70 (m, 1H), 7.50-7.30 (m, 1H), 6.20-5.80 (tt, J=56 Hz, J=4.4 Hz, 1H), 4.21 (s, 3H), 3.79 (s, 2H), 3.60-3.40 (m, 1H), 3.10-2.90 (m, 5H), 1.60-1.58 (d, 8H, J = 7.2 Hz). LCMS: 500.2 (M + H) ⁺ ; RT = 1.28 min

Example 14: 3-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7,8 -dihydro-1,6-naphthyridin-6(5H)-yl)propionitrile

N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7 was added sequentially to the dried reaction flask. , 8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (100 mg, 0.24 mmol), acrylonitrile (23.4 mg, 0.44 mmol), CuBr (12.6 mg, 0.08 mmol), After adding DMF (15 mL), water (5 mL), EtOAc (EtOAc) Dry over anhydrous sodium sulfate. Concentration by filtration afforded the crude compound which was crystallised eluted to afford 3-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidine-2- (Amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propanenitrile (10 mg, yield: 9.66%).

¹ H NMR (400 MHz, DMSO) δ 9.81 (s, 1H), 8.71 (s, 1H), 8.62.86. (d, 1H, J = 4 Hz), 8.11-8.09 (d, 1H, J = 8 Hz), 7.94 -7.92 (d, 1H, J = 8 Hz) 7.67-7.65 (d, 1H, J = 8 Hz), 7.45 - 7.43 (d, 1H, J = 8 Hz), 4.15 (s, 3H), 3.62-3.60 (d, 1H, J=8 Hz), 2.83 (s, 4H), 2.78 (s, 4H), 1.50-1.52 (d, 6H, J = 8 Hz), LCMS: 471.3 (M+H) ⁺ ; RT = 1.87 min.

The compound of Example 15 was prepared in the same manner as in Example 14.

Example 15: 3-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) -7,8-Dihydro-1,6-naphthyridin-6(5H)-yl)propanenitrile

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.49 (s, 1H), 8.39-8.38 (d, 1H, J = 4 Hz), 8.23 - 8.21 (d, 1H, J = 8 Hz), 8.01 (s, 1H), 7.76-7.73 (d, 1H, J = 12 Hz) 7.39-7.37 (d, 1H, J = 8 Hz), 4.20 (s, 3H), 3.70 (s, 2H), 3.51-3.47 (m, 1H), 2.97- 2.94 (m, 2H), 2.93-2.88 (m, 4H), 2.65-2.62 (m, 2H), 2.52 (s, 3H), 1.59-1.57 (d, 6H, J = 8 Hz), LCMS: 489.3 (M) +H) ⁺ ;RT=1.23min.

Example 16: (1-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7, 8-Dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone)

Add (N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7,8 to a 100 mL flask -tetrahydro-1,6-naphthyridin-2-amine) (80 mg, 0.19 mmol) and glycolic acid (30 mg, 0.38 mmol) to 15 mL of DMF, then DIEA (74 mg, 0.57 mmol), HBTU ( 144 mg, 0.38 mmol). The reaction was stirred at room temperature overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted with dichloromethane (3×50 mL). The organic phase was combined and dried over anhydrous sodium sulfate. Isolated product (1-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7,8 -Dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone) 38 mg, yellow powder, yield 41.70%.

¹ H NMR (400 MHz, DMSO) δ 9.87 (s, 1H), 8.70 (s, 1H), 8.62 (d, 1H, J = 3.6), 8.15 (d, 1H, J = 8.4 Hz), 7.92 (d, 1H, J=9.2Hz), 7.66 (d, 1H, J=9.2Hz), 7.54-7.61(m,1H), 4.62-4.68(m,2H), 4.18-4.22(m,2H), 4.15(s , 3H), 3.63-3.81 (m, 2H), 3.51-3.60 (m, 1H), 2.81-2.89 (m, 2H), 1.52 (d, 6H, J = 7.2 Hz). LCMS: 476.4 (M+H ) ⁺ ;RT=1.22min

The compound of Example 17, 18 was prepared in the same manner as in Example 16.

Example 17: 2-((5-Fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-6-(N-A ke-L-prolyl)-5,6,7,8-tetrahydro-1,6-diazepine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.80-8.60 (m, 1H), 8.50-8.30 (m, 2H), 8.10-7.90 (m, 2H), 7.80-7.70 (m, 1H), 7.60-7.20 ( m,1H), 4.90-4.60 (m, 2H), 4.22 (s, 3H), 4.10-3.80 (m, 2H), 3.60-3.50 (m, 1H), 3.30-3.20 (brs, 2H), 3.10- 2.90 (brs, 2H), 2.50-2.20 (m, 5H), 2.10 - 1.80 (m, 3H), 1.64-1.63 (d, 6H, J = 6.8 Hz). LCMS: 529.3 (M+H) ⁺ ; RT =1.11min

Example 18: (1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) )-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone)

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.42 (d, 1H, J = 5.2 Hz), 8.33 (d, 1H, J = 3.6), 8.25 (d, 1H, J = 8.4 Hz), 8.02-8.08 (m) , 1H), 7.63 (d, 1H, J = 12.8 Hz), 7.37-7.43 (m, 1H), 4.71 (s, 1H), 4.35 (s, 1H), 4.20-4.22 (m, 2H), 4.13 ( s, 3H), 3.91-3.94 (t, 1H, J = 6.0 Hz), 3.53-3.58 (m, 2H), 3.41-3.44 (m, 1H), 2.88-2.92 (m, 2H), 1.51 (d, 6H, J = 6.8 Hz). LCMS: 494.3, (M+H) ⁺ ; RT = 1.29 min.

Example 19: (6-(1-ethylpiperidin-4-yl)-N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl) Pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

Add (N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7,8 to a 100 mL flask -tetrahydro-1,6-naphthyridin-2-amine) (60 mg, 0.14 mmol) and 1-ethylpiperidin-4-one (37 mg, 0.286 mmol) in 20 mL of 1,2-dichloroethane After stirring at 60 ° C for half an hour, sodium triacetoxyborohydride (233 mg, 1.10 mmol) was added. The reaction was stirred at 60 ° C for 3 hours. After the reaction was completed, 50 mL of water was added, and then extracted with dichloromethane (3×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, Isolated product (6-(1-ethylpiperidin-4-yl)-N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidine 2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine) 15 mg, yellow powder, yield 19.72%.

¹ H NMR (400 MHz, DMSO) δ 9.79 (s, 1H), δ 8.70 (s, 1H), δ 8.61 (d, 1H, J = 2.0), δ 8.07 (d, 1H, J = 9.2) , δ 7.92 (d, 1H, J = 9.2), δ 7.66 (d, 1H, J = 9.2), δ 7.44 (d, 1H, J = 9.6), δ 4.15 (s, 3H), δ3 .67(s,2H),δ3.58-3.62(m,1H),δ2.96-3.17(m,4H),δ2.81-2.86(m,4H),δ2.66-2.67(m,1H) ), δ2.32-2.33(m, 2H), δ1.94-2.08(m, 2H), δ1.81-1.87 (m, 2H), δ1.51 (d, 6H, J=6.8), δ1. 01-1.03 (m, 3H). LCMS: 529.3 (M+H);

The compound of Example 20 was prepared in the same manner as in Example 19.

Example 20: N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(1- Methylpiperidin-4-yl)-5,6,7,8-tetrahydro-1,6-diazaphthalen-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.50 (s, 1H), 8.40 - 8.39 (d, 1H, J = 4 Hz), 8.20 - 8.18 (d, 1H, J = 8 Hz), 8.04 (s, 1H), 7.76-7.73 (d, 1H, J = 12 Hz), 7.38-7.36 (d, 1H, J = 8 Hz), 4.20 (s, 3H), 3.76 (s, 2H), 2.97-2.94 (d, 6H, J = 12 Hz), 2.55-2.46 (m, 1H), 2.29 (s, 3H), 2.04-1.97 (m, 2H), 1.93-1.89 (m, 2H), 1.71-1.74 (m, 2H), 1.478-1.569 ( d, 6H, J = 3.6 Hz), LCMS: 533.3 (M+H) ⁺ ; RT = 1.37 min

Example 21: (6-(2-(Dimethylamino)ethyl)-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-carbazole- 5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine)

Add (N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6 to a 100 mL flask ,7,8-tetrahydro-1,6-naphthyridin-2-amine) (90 mg, 0.21 mmol) and 2-bromo-N,N-dimethylethylamine (63 mg, 0.42 mmol) to 10 mL In DMF, DIEA (82 mg, 0.63 mmol) was then added. The reaction was stirred at 80 ° C overnight. After the reaction was completed, 50 mL of water was added, and the mixture was extracted twice with dichloromethane (2×50 mL). The organic phase was combined, dried over anhydrous sodium sulfate Chromatography to give the product (6-(2-(dimethylamino)ethyl)-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-carbazole) -5-yl)pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine) 18 mg, yellow powder, yield 17.19%.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.51 (s, 1H), 8.39 (d, 1H, J = 4.0), 8.27 (d, 1H, J = 8.4), 7.73 (d, 1H, J = 12.8), 7.41 (d, 1H, J = 8.8), 4.20 (s, 3H), 3.71 (s, 2H), 3.48-3.51 (m, 1H), 3.22-3.25 (m, 2H), 2.98-3.03 (m, 4H) ), 2.92-2.97 (m, 2H), 2.90 (s, 6H), 1.58 (d, 6H, J = 7.2). LCMS: 507.3 (M+H) ⁺ ; RT = 1.11 min

Example 22: 1-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7,8 -dihydro-1,6-naphthyridin-6(5H)-yl)propan-2-ol

N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-5,6,7 was added sequentially to the dried sealed tube. , 8-tetrahydro-1,6-naphthyridin-2-amine hydrochloride (40 mg, 0.088 mmol), water (1 mL), anhydrous potassium carbonate (42 mg, 0.3 mmol), THF (3 mL) - Methyl oxirane 5 (1 mL). The temperature was raised to 80 ° C for 0.5 hour. After completion of the reaction, the organic phase was separated and concentrated under reduced pressure. The crude product was purified by reverse-phase high performance liquid chromatography to give 1-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl) Amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-2-ol (13 mg, white solid).

¹ H NMR (400 MHz, CDCl3-d1) δ 8.70 (s, 1H), 8.38 (d, 1H, J = 5.2 Hz), 8.26 (d, 1H, J = 8.4 Hz), 8.03 (d, 1H, J = 9.2 Hz), 7.93 (s, 1H), 7.72 (d, 1H, J = 8.4 Hz), 7.35 (d, 1H, J = 8.4 Hz), 4.18 (s, 3H), 3.94 - 3.99 (m, 1H) , 3.80-3.83 (m, 1H), 3.56-3.60 (m, 1H), 3.47-3.54 (m, 1H), 3.03-3.07 (m, 1H), 2.94-2.96 (m, 2H), 2.80-2.84 ( m, 1H), 2.42 - 2.56 (m, 2H), 1.59 (d, 6H, J = 6.8 Hz), 1.20 (d, 3H, 1H, J = 6.4 Hz). LCMS: 476.3 (M+H)+; RT=0.873min.

Example 23: 1-(2-((5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) -7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-2-ol

¹ H NMR (400 MHz, CDCl3-d1) δ 8.50 (s, 1H), 8.40 (d, 1H, J = 4.0 Hz), 8.21. (d, 1H, J = 8.4 Hz), 8.06 (d, 1H, J = 7.6 Hz), 7.75 (d, 1H, J = 12.8 Hz), 7.37 (d, 1H, J = 8.4 Hz), 4.21 (d, 3H, J = 6.4 Hz), 3.95-3.99 (m, 1H), 3.82 (d, 1H, J = 14.4 Hz), 3.58 (d, 1H, J = 14.8 Hz), 3.46-3.53 (m, 2H), 3.03-3.08 (m, 1H), 2.96-2.97 (m, 2H), 2.78-2.83 (m, 1H), 2.42 - 2.56 (m, 2H), 1.58 (d, 6H, J = 7.2 Hz), 1.20 (d, 3H, J = 6.4 Hz). LCMS: 494.3 (M+H) +;RT=1.917min.

Example 24: 1-(2-((5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino) -7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-methylpropan-2-ol

¹ H NMR (400 MHz, CDCl3-d1) δ 8.50 (d, 1H, J = 1.2 Hz), 8.40 (d, 1H, J = 3.6 Hz), 8.20 (d, 1H, J = 8.4 Hz), 8.08 (s) , 1H), 7.75 (d, 1H, J = 12.8 Hz), 7.35 (d, 1H, J = 8.8 Hz), 4.20 (s, 3H), 3.81 (s, 2H), 3.02-3.06 (m, 3H) , 2.95-2.98 (m, 2H), 2.54 (s, 2H), 1.58 (d, 6H, J = 7.2 Hz), 1.24 (s, 6H). LCMS: 508.3 (M+H) +; RT = 1.860 min .

The compounds of Examples 25-28 were prepared in the same manner as in Example 1.

Example 25: 6-(2-ethoxyethyl)-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl) Pyrimidin-2-yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.50 (s, 1H), 8.41 - 8.40 (d, 1H, J = 4 Hz), 8.30 - 8.28 (d, 2H, J = 8 Hz), 7.75 - 7.72 (d, 1H) , J=12Hz), 7.43-7.41 (d, 1H, J=8Hz), 4.20(s, 3H), 4.08(s, 2H), 3.80-3.77 (q, 2H, J=12Hz), 3.57-3.55 ( d, 2H, J = 8 Hz), 3.54-3.49 (m, 1H), 3.30 (m, 2H), 3.11-3.10 (t, 3H, J = 4 Hz), 1.60-1.57 (d, 6H, J = 12 Hz) , 1.25-1.21 (t, 3H, J = 16 Hz), LCMS: 508.3 (M+H) ⁺ ; RT = 1.63.

Example 26: 6-(2,2-Difluoroethyl)-N-(5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidine-2 -yl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.69 (s, 1H), 8.40-8.30 (m, 1H), 8.29-8.20 (m, 1H), 8.10-8.00 (m, 1H), 7.95 (s, 1H) , 7.80-7.70 (m, 1H), 7.40-7.30 (m, 1H), 6.13-5.83 (tt, J = 56 Hz, J = 4.4 Hz, 1H), 4.17 (s, 3H), 3.77 (s, 2H) , 3.60-3.40 (m, 1H), 3.10-2.90 (m, 6H), 1.59-1.58 (d, 6H, J = 6.8 Hz), LCMS: 482.3 (M+H) ⁺ ; RT = 1.12 min

Example 27: N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-((tetrahydrofuran) -3-yl)methyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.50 (s, 1H), 8.39 - 8.38 (d, 1H, J = 4 Hz), 8.21 - 8.19 (d, 1H, J = 8 Hz), 7.97 (s, 1H), 7.76-7.73(d,1H,J=12Hz), 7.38-7.36(d,1H), 4.20(s,3H),3.93-3.88(m,2H),3.86-3.80(m,2H),3.78-3.76 (d, 2H, J = 8 Hz), 3.51-3.47 (m, 1H), 2.95-2.94 (t, 2H, J = 4 Hz), 2.86-2.85 (m, 2H), 2.60 (s, 3H), 2.10- 2.05 (m, 1H), 1.67 (s, 2H), 1.59-1.57 (d, 6H, J = 8 Hz), LCMS: 520.3 (M+H) ⁺ ; RT = 1.58 min.

Example 28: N-(5-Fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-(3- Methoxypropyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.62-8.60 (d, 1H, J = 8 Hz), 8.58 (s, 1H), 8.45-8.44 (d, 1H, J = 4 Hz), 7.71-7.68 (d, 1H) , J=12Hz), 7.65-7.63 (d, 1H, J=8Hz), 4.39(s, 2H), 4.21(s, 3H), 3.61(s, 2H), 3.52-3.48(m, 3H), 3.38 -3.34(m,2H),3.32-3.31(m,5H),2.13-2.11(m,2H), 1.59-1.57(d,6H,J=8Hz), LCMS:508.3(M+H) ⁺ ;RT =1.50min.

The compound of Example 29, 30 was prepared in the same manner as in Example 19.

Example 29: 6-Cyclopentyl-N-(5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl) -5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, DMSO) δ 10.91 (s, 1H), 10.09 (s, 1H), 8.64 (s, 1H), 8.54 (s, 1H,), 8.19 - 8.17 (d, 1H, J = 8 Hz) , 7.73-7.70 (d, 1H, J = 12 Hz), 7.64 - 7.62 (d, 1H, J = 8 Hz), 4.53 (s, 1H), 4.30 (s, 1H), 4.19 (s, 3H), 3.76- 3.72 (m, 1H), 3.72-3.63 (m, 2H), 3.05 (s, 1H), 2.11-2.07 (t, 3H, J = 16 Hz), 1.87-1.78 (m, 4H), 1.61 (s, 2H) ), 1.53-1.52 (d, 6H, J = 4 Hz), LCMS: 504.2 (M+H) ⁺ ; RT = 0.98 min.

Example 30: (N-(5-Fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)-6-methyl-5,6 ,7,8-tetrahydro-1,6-naphthyridin-2-amine

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.63 (s, 1H), 8.30-8.29 (t, 1H, J = 4 Hz), 8.20-8.17 (q, 1H, J = 12 Hz), 7.98-7.96 (d, 1H) , J=8Hz), 7.85(s,1H), 7.66-7.64(t,1H,J=8Hz), 7.28-7.20(br,s,1H),4.13(s,3H),3.53(s,2H) , 3.45-3.42 (m, 1H), 2.93 (s, 2H), 2.77 (s, 2H), 2.45 (s, 3H), 1.53-1.51 (d, 6H, J = 8 Hz), LCMS: 432.3 (M+ H) ⁺ ; RT = 1.71 min.

The compound of Example 31, 32 was obtained in the same manner as in Example 16.

Example 31: 2-((5-Fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)-7,8-dihydro -1,6-naphthyridin-6(5H)-yl)(1-methylpiperidin-4-yl)methanone

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.71 (s, 1H), 8.40-8.39 (d, 1H, J = 4 Hz), 8.36-8.34 (d, 1H, J = 8 Hz), 8.05-7.98 (br, s , 2H), 7.74-7.72 (d, 1H, J=8Hz), 7.48-7.46 (d, 1H, J=8Hz), 4.73-4.65 (m, 2H), 4.19 (s, 3H), 3.95-3.82 ( m, 2H), 3.56-3.49 (m, 1H), 2.98-2.94 (m, 4H), 2.56 (s, 1H), 2.31 (s, 3H), 2.05-1.91 (m, 3H), 2.60 (s, 3H), 2.10-2.05 (m, 1H), 1.78-1.73 (m, 3H), 1.61-1.59 (d, 6H, J = 8 Hz), LCMS: 543.3 (M+H) ⁺ ; RT = 1.10 min.

Example 32: 2-(Dimethylamino)-1-(2-((5-fluoro-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidine-2) -yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethan-1-one

¹ H NMR (400 MHz, CDCl ₃ ) δ 10.0-9.80 (m, 1H), 8.70 (s, 1H), 8.63 (s, 1H), 8.20-8.10 (m, 1H), 8.00-7.90 (m, 1H) , 7.70-7.60 (m, 1H), 7.59-7.50 (m, 1H), 4.80-4.50 (m, 2H), 4.14 (s, 3H), 3.90-3.80 (m, 2H), 3.70-3.50 (m, 2H), 3.28 (s, 2H), 3.00-2.70 (m, 2H), 2.25 (m, 6H), 1.52-1.50 (d, 6H, J = 6.8 Hz), LCMS: 503.3 (M+H) ⁺ ; RT=1.81min

Biological activity assay

1. Inhibitory activity test of the compound of the example against different CDK kinases (CDK1, CDK2, CDK4, CDK6, CDK9)

First step: In a 384-well plate, add 5 μL of enzyme system (50 mM HEPES, 10 mM MgCl ₂ , 1 mM EGTA, 2 mM DTT and 0.01% Tween 20, 0.1 μL enzyme), 2.5 μL of compound, 2.5 μL of substrate and ATP mix. The solution (final concentration of substrate was 50 nM, final concentration of ATP 100 nM) was incubated at room temperature for 60 min in the dark.

Step 2: Add 5 μL of 1× assay buffer diluted EDTA stop solution (final concentration 6 mM) to each well, then add 1× assay buffer diluted antibody (final concentration 2 nM) and incubate at room temperature for 1 h in the dark.

TRFRET mode测板。 Step 3: Using PerkinElmer En

TRFRET mode board.

The fourth step: IC50 values were calculated using GraphPad Prism software.

The test confirmed that the compound of the present invention has a good CDK4 inhibitory activity, and the inhibitory activity data of the compound of the present Example for CDK4 is shown in Table 1 below.

Table 1: CDK4 inhibitory activity of the compounds of the examples of the present application

Numbering CDK4/nM Numbering CDK4/nM 1 A 16 A 2 A 17 A 3 A 18 A 4 A 19 A 5 A 20 A 6 A twenty one A 7 A twenty two A 8 A twenty three A 10 A twenty four A 11 A 25 A 12 A 27 A 13 A 28 A 14 A 29 A 15 A 31 A

among them

A represents an IC ₅₀ of 1 nM to 50 nM;

B indicates an IC ₅₀ of 50 nM-100 nM

C indicates an IC ₅₀ of >100 nM

Furthermore, the experiments also confirmed that the compounds of the present invention have good selectivity for CDK4 relative to CDK1, i.e., the compounds of the present invention are selective inhibitors of CDK4. The selective inhibition data for exemplary compounds of the invention are shown in Table 2 below.

Table 2: Selective inhibition data for exemplary compounds of the invention

Numbering CDK1/nM CDK4/nM Numbering CDK1/nM CDK4/nM 1 550 1.7 16 307 1.8 2 995 2.3 18 198 1.8 3 1410 1.5 19 859 1.9

4 1467 2.9 20 2672 1.8 5 2058 1.9 twenty one 1272 1.9 6 7621 1.7 twenty two 218 1.6 7 10000 4.2 twenty three 585 1.3 8 10000 4.9 twenty four 10000 2.8 10 755 2.3 25 2358 4.0 11 10000 2.0 27 7789 3.1 12 525 1.3 28 2423 3.0 14 1680 1.9 29 10000 4.6 15 2382 1.6 31 431 2.1

2. Example compound test for proliferation inhibition activity of different tumor cells (Colo-205, MCF-7, MDA-MB468, LNCAP, K562, U87MG, HepG2, H1975, PANC-1) (CCK-8 or SRB method)

The specific steps of the CCK8 method are as follows: cells in the logarithmic growth phase are inoculated to a 96-well microplate at a suitable density, 200 μL per well, and after 24 hours of culture, different concentrations of compounds are added for 96 hr, and each concentration is set to three replicate wells. Corresponding concentrations of saline vehicle control and cell-free zeroing. After the end of the action, 20 μL of CCK-8 was added to each well, and cultured at 37 ° C for 1-4 hr, during which the color was observed, and the OD value was measured at a wavelength of 450 nm using a VERSMax plate reader. The inhibition rate of the drug on tumor cell growth was calculated by the following formula: inhibition rate (%) = (OD control well-OD administration well) / OD control well × 100%.

The specific steps of the SRB method are as follows: cells in the logarithmic growth phase are inoculated to a 96-well culture plate at a suitable density, 200 μL per well, and after incubation overnight, different concentrations of the drug are added for 96 hours, and each concentration is set to three replicate wells, and correspondingly Concentration of saline vehicle control and cell-free zeroing. After the end of the action, the adherent cells were decanted, and 10% (w/v) trichloroacetic acid (100 μL/well) was added and fixed at 4 ° C for 1 h, followed by rinsing with distilled water five times. After drying at room temperature, SRB solution was added to each well. (4mg/mL, dissolved in 1% glacial acetic acid) 100μL, incubate for 15min at room temperature, rinse with unwashed SRB five times with 1% glacial acetic acid, dry at room temperature, add 100μL of 10mM Tris solution per well, VERSMax enzyme The optical density (OD value) at a wavelength of 560 nm was measured by a spectrometer.

Numbering Colo-205/nM Numbering Colo-205/nM 1 72 16 213 2 105 18 158 3 58 19 137 4 525 20 231 5 355 twenty one 261

6 474 twenty two 95 7 1226 twenty three 121 8 1027 twenty four 7933 10 265 25 410 11 147 27 888 12 183 28 341 14 736 31 378 15 652

3. Determination of pharmacokinetic parameters of the compound of the example in rats and mice (boxed administration)

Six male SPF-class SD rats (Shanghai Shipu-Beikai experimental animals) were divided into two groups, and the test compound was configured into a suitable solution or suspension; one group was administered intravenously, and one group was orally administered.

Blood was collected by jugular vein puncture, and each sample was collected at about 0.2 mL/time. Heparin sodium was anticoagulated. The time of blood collection was as follows: before, and after 5, 15 and 30 min, 1, 2, 4, 6, and 8 after administration. 24 h; blood samples were collected and placed on ice, and plasma was separated by centrifugation (centrifugation conditions: 8000 rpm, 6 minutes, 2-8 ° C), and the collected plasma was stored at -80 ° C before analysis. Plasma samples were analyzed by LC-MS/MS. The time points of blood collection in brain tissue were: 0.5, 2, 6 h (one time point for every 3 animals).

After the animal was euthanized, blood was collected by cardiac puncture for about 0.50 mL, and then the brain tissue was collected and collected and placed on ice. After the animals are euthanized, brain tissue is collected. The tissue was washed with physiological saline, the filter paper was blotted dry, weighed, and the scissors were cut and placed in an EP test tube. The EP tube was placed in a fully automatic homogenizer, homogenized by adding 50% methanol 50% water according to a corresponding volume of 5 mL/g, and each tissue homogenate was frozen at -20 ° C for testing.

According to the blood concentration data of the drug, the pharmacokinetic parameters ANU _0-t , AUC _0-∞ , MRT _0-∞ , C _{max of the} test sample were calculated using the non-compartment model of the pharmacokinetic calculation software WinNonlin5.2. Parameters such as T _max , T _1/2 and V _d and their mean and standard deviation. In addition, the bioavailability (F) will be calculated by the following formula.

For samples with concentrations below the lower limit of quantitation, samples taken prior to reaching C _max should be calculated as zero values when calculating the pharmacokinetic parameters. Samples at the sampling point should be incapable of quantification (BLQ) after C _{max is} reached.

The main pharmacokinetic parameters of plasma in single injection and oral administration of SD rats after Example 23

Plasma and brain tissue concentrations after oral administration of Example 23 in SD rats

Experimental conclusion: The representative compound of the present invention (Example 23) has good PK properties in mouse and rat experiments and has a good blood-brain distribution. It is suggested that the compounds of the invention can cross the blood brain barrier to inhibit the growth of brain tumors.

Forty-eight male SPF-class ICR mice were purchased from Shanghai Xipuer-Beikai Experimental Animal Co., Ltd., and 48 healthy ICR mice with good physical examination and no abnormality were used for the study. The test compound is formulated as a suitable solution or suspension; one group is administered intravenously and the other is administered orally.

After CO2 euthanasia, blood was collected by cardiac puncture. Each sample was collected at about 0.20 mL/time point. Heparin sodium was anticoagulated and placed on ice after collection. The time of blood collection was as follows:

Intravenous group: 5, 15 and 30 min, 1, 4, 8 and 24 h before and after administration.

Oral administration group: before administration and 30 minutes after administration, 1, 2, 4, 6, 8 and 24 hours.

Blood samples were collected and placed on ice, and plasma was separated by centrifugation (centrifugation conditions: 8000 rpm, 6 minutes, 2-8 ° C). The collected plasma was stored at -80 °C prior to analysis.

Plasma samples were analyzed by LC-MS/MS by the laboratory analysis department. According to the blood drug concentration data of the drug, the pharmacokinetic parameters ANU0-t, AUC0-∞, MRT0-∞, Cmax, Tmax, T1 of the test sample were calculated using the non-compartment model of the pharmacokinetic calculation software WinNonlin7.0. Parameters such as /2 and Vd and their mean and standard deviation.

Single injection and oral administration of the main pharmacokinetic parameters of Example 23

4. Test of inhibition of growth of transplanted tumor in nude mice by the compound of the example

The tumor tissue in the vigorous growth period was cut into 1.5 mm ³ and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions. The nude mice were transplanted subcutaneously with a vernier caliper to measure the diameter of the transplanted tumor. The animals were randomly divided into tumors with an average volume of about 130 mm ³ . The compound of the example (administered to the desired concentration with water for injection containing 1% Tween 80) was orally administered at a given dose per day for three weeks, and the solvent control group was given an equal amount of solvent. The diameter of the transplanted tumor was measured twice a week during the entire experiment, and the body weight of the mice was weighed. The formula for calculating tumor volume (TV) is: TV = 1/2 × a × b ² , where a and b represent length and width, respectively. Based on the measured results, the relative tumor volume (RTV) was calculated, and the formula was: RTV=V _t /V ₀ . Where V ₀ is the measured tumor volume at the time of sub-cage administration (i.e., d ₀ ), and V _t is the tumor volume at each measurement. The evaluation index of antitumor activity is 1) relative tumor proliferation rate T/C (%), and the formula is as follows: T/C (%) = (T _RTV / C _RTV ) × 100%, T _RTV : treatment group RTV; C _RTV : negative control RTV; 2) tumor volume growth inhibition rate GI%, calculated as follows: GI% = [1-(TVt-TV ₀ ) / (CVt-CT ₀ )] × 100%, TVt for the treatment group Secondary measured tumor volume; TV ₀ is the tumor volume obtained when the therapeutic component is administered in cage; CVt is the tumor volume measured in each control group; CV ₀ is the tumor volume obtained when the cage is administered in the control group; 3) Tumor weight inhibition The rate was calculated as follows: tumor weight inhibition rate % = (Wc - W _T ) / Wc × 100%, Wc: control group tumor weight, WT: treatment group tumor weight.

Example 23 Experimental treatment of human colon cancer COLO 205 nude mice xenografts

Example 23 was administered orally at a dose of 100 mg/kg and 30 mg/kg once a day for 21 days, which significantly inhibited the growth of subcutaneous xenografts of human colon cancer COLO 205 nude mice. At LY2835219.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (13)

A compound of the formula I, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof,

among them, R _{1 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₂ -C ₆ alkenyl, optionally substituted C ₂ -C ₆ alkynyl, optionally substituted C ₃ - a C ₈ cycloalkyl group, an optionally substituted 4-8 membered heterocyclic group containing 1, 2 or 3 hetero atoms selected from nitrogen, oxygen and sulfur, an optionally substituted carbonyl group, an optionally substituted sulfonyl group, and An optionally substituted sulfinyl group; R _{2 is} selected from the group consisting of: halogen, -OH, -CN, -NR _a R _b , optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted C ₃ a -C ₈ cycloalkyl group, and an optionally substituted 3-8 membered heterocyclic group; R _a and R _b are each independently selected from the group consisting of: H, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, and hydroxy substituted C ₁ -C ₆ alkyl; m is the number of R ₂ substituents and is 0, 1, 2 or 3; R ₃ is hydrogen or halogen; R ₄ is an optionally substituted C ₁ -C ₆ alkyl group; R _{5 is} selected from the group consisting of hydrogen, an optionally substituted C ₁ -C ₆ alkyl group, an optionally substituted 4-8 membered heterocyclic group, and an optionally substituted C ₃ -C ₈ cycloalkyl group. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, characterized in that Yes, R _{1 is} selected from the group consisting of: an optionally substituted C ₁ -C ₆ alkyl group, an optionally substituted C ₂ -C ₆ alkenyl group, an optionally substituted C ₂ -C ₆ alkynyl group, an optionally substituted C ₃ -C ₈ a cycloalkyl group, an optionally substituted 4-8 membered heterocyclic group, an optionally substituted C ₁ -C ₆ alkylcarbonyl group, an optionally substituted 4-8 membered heterocyclic carbonyl group, an optionally substituted sulfonyl group, and An optionally substituted sulfinyl group; the optional substituent described in R ₁ is one or more substituents independently selected from the group consisting of halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkane a hydroxy-substituted C ₁ -C ₄ alkyl group, a C ₁ -C ₃ alkoxy group, a -NH(C ₁ -C ₄ alkyl group), a -N(C ₁ -C ₄ alkyl) ₂ group, a C ₃ group C ₈ cycloalkyl and optionally substituted by C ₁ -C ₄ alkyl, halo C ₁ -C ₄ alkyl or hydroxy substituted C ₁ -C ₄ alkyl, containing 1, 2 or 3 selected from nitrogen a 4-8 membered heterocyclic group of oxygen and sulfur; R _{2 is} selected from the group consisting of: halogen, -OH, -CN, -NR _a R _b , optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₁ -C ₆ alkoxy, optionally substituted 4- An 8-membered heterocyclic group, and an optionally substituted C ₃ -C ₇ cycloalkyl group; the optional substituents described in R ₂ are one or more substituents independently selected from the group consisting of halogen, -OH, - NH ₂ , -CN, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₄ alkyl), and -N(C ₁ -C ₄ alkyl) ₂ ; R _a and R _b are each independently selected from the group consisting of: H, C ₁ -C ₄ alkyl, halogenated C ₁ -C ₄ alkyl, and hydroxy-substituted C ₁ -C ₄ alkyl; m is the number of R ₂ substituents and is 0, 1, 2 or 3; R ₃ is hydrogen or halogen; R ₄ is C ₁ -C ₄ alkyl; R ₅ is selected from: hydrogen, optionally substituted C ₁ -C ₆ alkyl, optionally substituted C ₄ -C ₈ heterocyclyl, and optionally substituted C ₃ -C ₈ cycloalkyl group; R ₅ as said optional substituents are one or more substituents independently selected from the group consisting of: _{halo, -OH, -CN, C 1 -C} 4 alkyl, and C ₃ -C ₆ cycloalkyl. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, characterized in that Yes, R _{1 is} selected from the group consisting of: an optionally substituted C ₁ -C ₄ alkyl group, an optionally substituted C ₃ -C ₆ cycloalkyl group, an optionally substituted 4-6 membered heterocyclic group, an optionally substituted C ₁ -C _{a 4} -alkylcarbonyl group and an optionally substituted 4-6 membered heterocyclylcarbonyl group; the optional substituents described in R ₁ are one or more substituents independently selected from the group consisting of halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkyl, hydroxy substituted C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ - C ₂ alkyl) _2, C ₃ -C ₆ cycloalkyl, and optionally substituted C ₁ -C ₂ alkyl, halo-C ₁ -C ₃ alkyl or hydroxy-substituted C ₁ -C ₃ alkyl substituted with a 4-6 membered heterocyclic group containing 1 or 2 selected from the group consisting of nitrogen, oxygen and sulfur; Preferably, R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, 4-6 membered heterocyclic, C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclic group a hydroxy-substituted C ₁ -C ₃ alkyl-substituted 4-6 membered heterocyclic group, a halogenated C ₁ -C ₃ alkyl-substituted 4-6 membered heterocyclic group, a hydroxy-substituted C ₁ -C ₂ alkane Alkylcarbonyl; R _{1a is} selected from the group consisting of: halogen, -OH, -NH ₂ , -CN, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane ₂ , C ₃ -C ₆ cycloalkyl, 4-6 membered heterocyclyl, C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclyl, hydroxy substituted C ₁ -C ₂ alkyl substituted a 4-6 membered heterocyclic group, and a halogenated C ₁ -C ₂ alkyl substituted 4-6 membered heterocyclic group, the heterocyclic group being a heterocyclic group containing 1 or 2 selected from nitrogen and oxygen; More preferably, R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, hydroxymethyl substituted carbonyl,

R _{1a is} selected from the group consisting of: F, -OH, -NH ₂ , -CN, CF ₃ , C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane Base) ₂ ,

R _{1b is} selected from the group consisting of methyl, ethyl and hydroxy substituted propyl groups; R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate or prodrug thereof, wherein R _{2 is} selected from the group consisting of: halogen, -OH, -NH ₂ , -CN, C ₁ -C ₄ alkyl, m is the number of R ₂ substituents, and is 0, 1; preferably, R _{2 is} selected from: F, Cl , -OH, -NH ₂ , -CN, C ₁ -C ₃ alkyl, m is the number of R ₂ substituents, and is 0, 1; more preferably, m is the number of R ₂ substituents, and is 0 . A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, characterized in that In the case, R ₃ is H or F. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, characterized in that Wherein R ₄ is a C ₁ -C ₂ alkyl group; preferably, R ₄ is a methyl group. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, characterized in that Yes, R _{5 is} selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₄ alkyl, and optionally substituted C ₃ -C ₆ cycloalkyl; the optional substituents described in R ₅ are one or two independent a substituent selected from the group consisting of halogen, -OH, -CN, C ₁ -C ₄ alkyl, and C ₃ -C ₅ cycloalkyl; Preferably, R _{5 is} selected from:

More preferably, R ₅ is

A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate or prodrug thereof, wherein R _{1 is} selected from the group consisting of R _1a (C ₁ -C ₄ alkyl), C ₃ -C ₆ cycloalkyl, hydroxymethyl substituted carbonyl,

R _{1a is} selected from the group consisting of: F, -OH, -NH ₂ , -CN, CF ₃ , C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₂ alkyl), -N(C ₁ -C ₂ alkane Base) ₂ ,

R _{1b is} selected from the group consisting of methyl, ethyl and hydroxy substituted propyl groups; R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl; m is the number of R ₂ substituents and is 0; R ₃ is hydrogen or F; R ₄ is a methyl group; R ₅ is

A compound according to claim 8, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate or prodrug thereof, according to claim 8, R _{1 is} selected from the group consisting of: R _1a (C ₁ -C ₄ alkyl),

R _{1a is} selected from the group consisting of: F, -OH, CF ₃ , C ₁ -C ₃ alkoxy, -NH ₂ , -N(C ₁ -C ₂ alkyl) ₂ ,

R _{1b is} selected from the group consisting of: methyl, and ethyl; R _{1aa is} selected from the group consisting of: C ₁ -C ₂ alkyl. A compound according to claim 1, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or prodrug thereof, wherein The compound is selected from

A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 10, or a stereoisomer, geometric isomer, tautomer thereof, pharmaceutically Acceptable salts, crystal forms, solvates, hydrates or prodrugs, as well as pharmaceutically acceptable excipients. A compound according to any one of claims 1 to 10, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, crystal form, solvate, hydrate or former thereof The use of a medicament for the manufacture of a medicament for preventing, treating, or alleviating a disorder or disease caused by abnormal cell proliferation, autoimmunity, inflammation or infection in a patient. The use according to claim 12, wherein the abnormal cell proliferation, autoimmunity, inflammation or infection is caused by a change in a cyclin-dependent kinase.