CN109970717B

CN109970717B - 4- (alicyclic pyrimidine/pyridine substituted) amino-1H-3-pyrazolecarboxamide FLT3 inhibitor and application thereof

Info

Publication number: CN109970717B
Application number: CN201711499769.2A
Authority: CN
Inventors: 王越; 卢帅; 支燕乐; 尧超; 陆涛; 李保泉; 陈璞洲; 鲍吉银
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2017-12-28
Filing date: 2017-12-28
Publication date: 2022-10-18
Anticipated expiration: 2037-12-28
Also published as: CN109970717A

Abstract

The invention relates to a 4- (saturated fat ring pyrimidine/pyridine substituted) amino-1H-3-pyrazole carboxamide FLT3 inhibitor and application thereof or pharmaceutically acceptable salt, solvate, isomer, ester, acid, metabolite or prodrug thereof, a preparation method thereof, a pharmaceutical composition containing the compound, and medical application thereof.

Description

4- (alicyclic pyrimidine/pyridine substituted) amino-1H-3-pyrazolecarboxamide FLT3 inhibitor and application thereof

Technical Field

The present invention relates to novel FLT3 kinase inhibitor compounds, pharmaceutical compositions comprising the compounds, and uses and methods of using the compounds and compositions to reduce or inhibit FLT3 kinase and/or mutant FLT3 kinase activity in a cell or a subject, and to prevent or treat a cell proliferative disorder and/or a FLT 3-related disorder in a subject.

Background

Cell signal transduction plays a key role in regulating the processes of cell growth, proliferation, differentiation, apoptosis and the like. Imbalance between cell proliferation and apoptosis leads to the development of serious diseases such as cancer, and the essence of cell canceration is the dysregulation of cell signal transduction. When the cell signal transduction pathway regulating the normal physiological activities of cells in the cells is changed under the action of carcinogenic factors, the normal biological effects regulating the growth, division and differentiation of the cells are abnormal, so that the abnormal growth and division of the cells and the change of cell morphology are caused, and the occurrence of cancers is caused. Because the protein kinase plays a key role in a cell signal transduction pathway and the cancer cells are often accompanied with the over-expression of the protein kinase, the protein kinase has wide prospect for developing the anti-tumor drugs by taking the protein kinase as a target point.

In recent years, protein kinase inhibitors have been a hot spot in the research and development of antitumor drugs, and many small molecule kinase inhibitors such as imatinib, sorafenib and sunitinib have been marketed, and in addition, a large number of small molecule kinase inhibitors are in clinical research. FMS-like tyrosine kinase 3 (FLT3) is a type III receptor tyrosine kinase and plays a key role in the proliferation, differentiation and apoptosis of hematopoietic cells and lymphocytes. Mutant FLT3 generally exhibits higher activity and is capable of autophosphorylation activation of downstream signaling pathways without ligand binding. These pathways include the RAS/MEK, PI3K/AKT/mTOR, and JAK/STAT5 pathways. FLT3 receptors are widely distributed in various tissues such as bone marrow hematopoietic stem/progenitor cells, thymus, lymph, placenta, brain, gonads, and the like. Abnormal activation of FLT3 is closely associated with the development of a range of hematological malignancies, especially Acute Myeloid Leukemia (AML). Therefore, the development of inhibitors targeting FLT3 is a hot research focus for the treatment of hematological malignancies.

Hematologic malignancies are cancers of the body's blood formation and immune system, bone marrow and lymphoid tissues. Although FLT3 expression is restricted to early progenitor cells in normal bone marrow, FLT3 expression at high levels or FLT3 mutations cause uncontrolled FLT3 receptors and downstream molecular channels to induce activation of possibly RAS, JAK, PI3K, etc. in hematological malignancies. Hematological malignancies include leukemia, lymphoma (non-hodgkin lymphoma), hodgkin's disease (also known as hodgkin lymphoma), and myeloma, for example, acute Lymphocytic Leukemia (ALL), acute myelocytic leukemia or Acute Myeloid Leukemia (AML), acute Promyelocytic Leukemia (APL), chronic Lymphocytic Leukemia (CLL), chronic Myelocytic Leukemia (CML), chronic Neutrophilic Leukemia (CNL), acute undifferentiated cell leukemia (AUL), operational developmental large cell lymphoma (ALCL), adult T cell ALL, AML with lan lineage (triline) myelodysplasia (AML/s), mixed Lineage Leukemia (MLL), myelodysplastic syndrome (mds), myelodysplasia (MPD), multiple Myeloma (MM), and myxosarcoma.

FLT3 plays a key role in hematopoiesis and lymphocyte proliferation, and abnormal activation of FLT3 is closely related to the occurrence and development of various tumors, particularly Acute Myelogenous Leukemia (AML). AML patients with high FLT3 expression still have a high recurrence rate even after bone marrow transplantation, making the prognosis of the tumor poor. At present, targeted inhibition of FLT3 and mutant FLT3 becomes a research hotspot, mainly aims to develop a small-molecule tyrosine kinase inhibitor, and inhibits the activity of the inhibitor by competing ATP binding sites with FLT3 tyrosine kinase. Kinase inhibitors that inhibit FLT3 that have been clinically developed at present include PKC412, AC220, and the like. To date, small molecule FLT3 inhibitors have been explored in the treatment of AML patients, and although some small molecule FLT3 inhibitors are currently in clinical research, the therapeutic efficacy is still very limited. This is because the high degree of homology among the various tyrosine kinase receptor structures results in poor inhibitor selectivity, and thus it is difficult to achieve an effective dose for inhibiting FLT3 in AML patients over a range of drug-resistant amounts. On the other hand, the occurrence of resistance mutations is also an important factor affecting the treatment of FLT3 inhibitors. In studies on highly heterogeneous AML, FLT3 mutations were the 1 st found. FLT3 has high homology with other kinases, and the existing FLT3 inhibitor also has an inhibitory effect on other kinases, so that the FLT3 inhibitor with high activity and good selectivity has great significance for researching the relation between FLT3 and diseases and treating related diseases.

Until now, the selective FLT3 inhibitor AC220 developed by Daiichi Sankyo corporation entered phase three clinical studies, and no selective FLT3 inhibitor has been marketed. Currently, many FLT3 inhibitors are in clinical research phase, and still more FLT3 inhibitors are in preclinical drug discovery phase.

Disclosure of Invention

The compound has excellent FLT3 selectivity and inhibition activity, and shows stronger antitumor activity. Which includes compounds of formula (I) or a pharmaceutically acceptable salt, solvate, isomer, ester, acid, metabolite or prodrug thereof:

the technical scheme of the invention is as follows:

a compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is ¹ Represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, heterocycloalkyl, alkylamino, alkoxyalkyl, alkoxyformyl, alkylcarbamoyl, aralkyl, diarylalkyl, arylheterocycloalkyl, aryl or heteroaromatic;

x and Y each independently represent an N atom or a CH atom group; wherein the CH radical may optionally be substituted by R ² Substituted, R ² Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

A ¹ represents NH, O, S or an alkylene radical; wherein each NH radical or alkylene radical independently may be optionally substituted by R ³ Substituted, R ³ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

A ² each independently represents a bond, alkylene, C (O) NH, C (O), NHC (O), alkylene-C (O), C (O) -alkylene, alkylene-C (O) -alkylene, or NHC (O) NH; wherein alkylene, C (O) NH, NHC (O), alkylene-C (O), C (O) -alkylene, alkylene-C (O) -alkylene or NHC (O) NH each independently may optionally be substituted by R ⁴ Substituted, R ⁴ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxyA group, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic ring;

Q ¹ is selected from aryl or aromatic heterocycle, wherein aryl or aromatic heterocycle independently of each other may optionally be substituted by one or more R ⁵ Substituted, R ⁵ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

Q ² is selected from aryl, aromatic heterocycle, C ₃ -C ₈ Aliphatic carbocycle, heterocycloalkyl, -NH of ₂ or-OH, wherein aryl, aromatic heterocycle, C ₃ -C ₈ Each independently of the other, may be optionally substituted with one or more R ⁶ Substituted, R ⁶ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic; -NH ₂ or-OH independently of one another may optionally be substituted by one or more R ⁷ Substituted, R ⁷ Can be hydrogen, alkyl, haloalkyl, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

the value range of n is 1-4;

alkyl is a straight or branched chain saturated or partially unsaturated hydrocarbon group having 1 to 8 carbon atoms; or a cyclic saturated or partially unsaturated hydrocarbon group having 3 to 8 carbon atoms; or a cyclic saturated or partially unsaturated hydrocarbon group having 3 to 8 carbon atoms to which a linear or branched saturated or partially unsaturated hydrocarbon group having 1 to 8 carbon atoms is attached;

alkylene is a straight or branched chain saturated hydrocarbon group having 1 to 8 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms; or a group formed by leaving one hydrogen atom of a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms to which a straight or branched chain saturated hydrocarbon group having 1 to carbon atoms is bonded;

heterocycloalkyl is a saturated or partially unsaturated monocyclic or polycyclic cyclic alkyl substituent comprising 3 to 12 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or a heteroatom of S (O) m (where m is an integer from 0 to 2), the remaining ring atoms are carbon, each monocyclic or bicyclic ring being optionally substituted with 1, 2, or 3 substituents, each substituent being independently selected from halogen, haloalkyl, hydroxy, alkyl, or alkoxy; halogen is a substituent selected from fluorine, chlorine, bromine or iodine;

alkoxy is a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms to which a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms is attached; wherein each carbon atom is optionally substituted by oxygen;

alkylthio is a straight or branched chain saturated hydrocarbon radical having 1 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms to which a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms is bonded; wherein each carbon atom is optionally substituted with sulfur;

alkylamino is a straight or branched chain saturated hydrocarbon radical having 1-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms; or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms to which a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms is bonded; wherein each carbon atom is optionally substituted with an NH radical;

alkoxyalkyl is an alkoxy group as defined above attached to an alkyl group;

aryl is a carbocyclic ring selected from phenyl, naphthyl, acenaphthyl or tetrahydronaphthyl, each of which is optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl or heteroaromatic ring;

the aromatic heterocyclic ring is monocyclic heterocyclic ring selected from pyrrolyl, pyrazolyl, imidazolyl, furyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl and pyridazinyl; or a bicyclic heterocycle selected from the group consisting of quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2, 3-dihydrobenzo [1,4] dioxinyl, benzo [1,3] dioxolyl;

aralkyl, diarylalkyl, arylheterocycloalkyl, or arylheterocycle as defined above attached to an alkyl group;

aryloxy, arylamino, arylthio, aralkyloxy, aralkylamino, aralkylthio are aryl groups as defined above linked to O, S, NH, alkoxy, alkylamino, alkylthio, respectively;

the haloalkyl group is a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms, or a cyclic saturated hydrocarbon group having 3 to 6 carbon atoms to which a straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms is attached; wherein one or more carbon atoms are substituted with one or more halogen atoms.

The preferred scheme of the invention is as follows:

R ¹ represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkylamino, heterocycloalkyl, alkoxyalkyl, alkoxyformyl or alkylcarbamoyl;

x, Y and Z independently represent N atom or CH atom group; wherein the CH radical may optionally be substituted by R ² Substituted, R ² Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

A ¹ each independently represents NH, O, S or an alkylene radical; wherein each NH radical or alkylene radical independently may be optionally substituted by R ³ Substituted, R ³ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

A ² each independently represents a bond, alkylene, C (O) NH, C (O), NHC (O), alkylene-C (O), C (O) -alkylene, alkylene-C (O) -alkylene, or NHC (O) NH; wherein alkylene, C (O) NH, NHC (O), alkylene-C (O), C (O) -alkylene, alkylene-C (O) -alkylene or NHC (O) NH each independently may optionally be substituted by R ⁴ Substituted, R ⁴ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxyA group, alkylthio or alkoxyalkyl;

Q ¹ is selected from aryl or aromatic heterocycle, wherein aryl or aromatic heterocycle independently of each other may be optionally substituted by one or more R ⁵ Substituted, R ⁵ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

the value range of n is 1-4.

Another preferred embodiment of the present invention is:

wherein R is ¹ Represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxyl, mercapto, alkoxy, alkylthio or alkylamino;

x and Y each independently represent an N atom or a CH atom group;

A ¹ each independently represents NH, O, S or an alkylene radical;

A ² each independently represents a bond, alkylene, C (O) NH, C (O), NHC (O), alkylene-C (O), C (O) -alkylene, alkylene-C (O) -alkylene, or NHC (O) NH;

Q ¹ is selected from aryl or aromatic heterocycle, wherein aryl or aromatic heterocycle independently of each other may optionally be substituted by one or more R ⁶ Substituted, R ⁶ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

Q ² is selected from aryl, aromatic heterocycle, C ₃ -C ₈ Aliphatic carbocycle, heterocycloalkyl, -NH of ₂ or-OH, wherein aryl, aromatic heterocycle, C ₃ -C ₈ Each independently of the other, may be optionally substituted with one or more R ⁷ Substituted, R ⁷ Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic; -NH ₂ or-OH independently of one another may optionally be substituted by one or more R ⁸ Substituted, R ⁸ Can be hydrogen, alkyl, haloalkyl, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

the value range of n is 1-4.

Another preferred embodiment of the present invention is:

wherein R is ¹ Selected from hydrogen; or an alkyl group selected from: methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl; or a heterocycloalkyl group of: oxetanyl, oxocyclopentyl, oxocyclohexyl, tetrahydropyrrolyl, piperidinyl, N-methylpiperidin-4-yl, morpholinyl, N-methylpiperazinyl, 3-methylpiperidin-1-yl, piperazinyl; or alkoxy and alkylamino of: cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, cyclopentylmethoxy, cyclopentylethoxy, cyclopropylamino, cyclobutylamino, cyclopentylamino, cyclohexylamino, cycloheptylamino; or the following aryl and heteroaromatic rings: phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, pyrrolyl, furanyl, thienyl, pyridyl, pyrimidinyl; or the following aryloxy and arylamino groups: phenoxy, naphthoxy, anilino, naphthylamino; or the following aralkyloxy and aralkylamino groups: benzyloxy, benzylamino;

x and Y independently represent N atom or CH atomic group, and at least one of X and Y is CH atomic group;

A ¹ represents NH;

A ² each independently represents a bond, alkylene, or C (O);

Q ¹ is an aromatic or substituted aromatic ring selected from: phenyl, naphthyl, pyrrolyl, furyl, thienyl, pyridyl, pyrazinyl and pyrimidinyl, wherein the substituent can be 1-2 halogens, methyl, methoxy or trifluoromethyl;

Q ² is an aliphatic heterocycle selected from: tetrahydropyrrolyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, thiomorpholinyl, pyranyl, tetrahydrofuranyl, aziridinyl, azetidinyl, N-methylpiperidin-4-yl, N-methylpiperazinyl, 3-methylpiperidin-1-yl, piperazinyl; or substituted amino, substituted oxy selected from: n, N-dipropylamino, N-diethylamino, N-dimethylamino, N-butylamino, 2-methoxyethoxy, 2-hydroxyethylamino, N-di (2-methoxyethyl) amino;

the value range of n is 1-4.

Another preferred embodiment of the present invention is:

wherein R is ¹ Represents hydrogen, methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclohexyl or cyclopentyl;

x, Y and Z independently represent N atom or CH atom group, and at least one of X, Y and Z is CH atom group;

A ¹ represents NH;

A ² each independently represents a bond or CH ₂ ；

Q ¹ Represents a phenyl group;

Q ² is selected from: tetrahydropyrrolyl, morpholinyl, N-methylpiperazinyl, piperazinyl;

the value range of n is 1-4.

According to the invention, pharmaceutically acceptable salts include the acid addition salts of the compounds of formula I with the following acids: hydrochloric, hydrobromic, sulphuric, phosphoric, methanesulphonic, benzenesulphonic, p-toluenesulphonic, naphthalenesulphonic, citric, tartaric, lactic, pyruvic, acetic, maleic or succinic, fumaric, salicylic, phenylacetic, mandelic acid. Also included are acid salts of inorganic bases such as: contains alkali metal cation, alkaline earth metal cation, and ammonium cation salt.

The compounds of formula I are preferably compounds of the following structure:

4- ((6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-1)

N- (4- (piperazin-1-yl) phenyl) -4- ((5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-2)

4- ((6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (morpholinomethyl) phenyl) -1H-pyrazole-3-carboxamide (I-3)

N- (4- (morpholinomethyl) phenyl) -4- ((5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-4)

4- ((2-methyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (morpholinomethyl) phenyl) -1H-pyrazole-3-carboxamide (I-5)

4- ((2-methyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (morpholinomethyl) phenyl) -1H-pyrazole-3-carboxamide (I-6)

4- ((2-methyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-7)

4- ((2-methyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-8)

4- ((2-methyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-pyrazole-3-carboxamide (I-9)

4- ((2-methyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-pyrazole-3- -carboxamide (I-10)

N- (4- (piperazin-1-yl) phenyl) -4- ((2- (trifluoromethyl) -6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-11)

N- (4- (piperazin-1-yl) phenyl) -4- ((2- (trifluoromethyl) -5,6,7, 8-tetrahydroquinazolin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-12)

4- ((2-isopropyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-13)

4- ((2-isopropyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-14)

4- ((2-Ethyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-15)

4- ((2-Ethyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-16)

4- ((2-chloro-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-17)

4- ((2-chloro-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-18)

4- ((2-cyclopropyl-6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-19)

4- ((2-cyclopropyl-5, 6,7, 8-tetrahydroquinazolin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-20)

4- ((2-methyl-6, 7,8, 9-tetrahydro-5H-cyclohepta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-21)

4- ((2-methyl-6,7,8,9-tetrahydro-5H-cyclohepta [ d ] pyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-22).

The preparation method of the partial compound of the invention comprises the following steps:

the method comprises the following steps:

the second method comprises the following steps:

the third method comprises the following steps:

the compound of the invention can be prepared by the preparation method or similar preparation methods, and corresponding raw materials are selected according to different substituents and different positions of the substituents.

Pharmacological test results show that the compound shown in the general formula I and the pharmaceutically acceptable salt thereof have excellent inhibitory activity on FLT3, so that the compound shown in the general formula I and the pharmaceutically acceptable salt thereof can be used for treating clinical diseases related to the kinase. Diseases related to the above kinases may be, but are not limited to: lung cancer, melanoma, liver cancer, kidney cancer, leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gall bladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, thyroid follicular cancer, gastrointestinal cancer, tumors of the central or peripheral nervous system (e.g., astrocytoma, neuroblastoma, glioma or schwannoma), mesothelioma, type II or non-insulin dependent diabetes mellitus, autoimmune diseases. (1) Determination of kinase inhibitory Activity of target Compound

The synthesized compounds were measured for inhibitory activity against FLT3, CDK2, 4, 6 by Fluorescence Resonance Energy Transfer (FRET) method, respectively, and compounds with better activity were screened compared with positive control drugs. The above kinases were obtained by purification or direct purchase of kits. Taking the test of the inhibitory activity of FLT3 as an example, the specific method is as follows:

FLT3 was diluted to an appropriate concentration with a kinase diluent and used. The kinase reaction mixture contains FLT3, peptide substrate, HEPES (pH 7.5), BRIJ-35, mgCl ₂ And EDTA. CDK2phospho-peptide substrate was used as 100% phosphorylation control and no ATP was used as 0% phosphorylation control. After 1h at room temperature, development Reagent A was added to the reaction system at moderate dilution. At room temperatureThe reaction is continued for 1h, and Stop Reagent is added to Stop the reaction. Excitation wavelength 400m, while fluorescence intensities at 445nm (coumarins) and 520nm (fluorosceins) were detected. The inhibition rate of the test compound is calculated according to the formula.

(2) Inhibitory Activity of Compounds on partial kinases (inhibition Rate%, 1X 10) ^-6 mol/L)

(3) In vitro anti-tumor Activity assay for Compounds of interest

MTT method is used to determine the inhibition effect on gastric cancer cell strain MGC803, leukemia cell strain K562, breast cancer cell strain MCF7, leukemia cell strain MV4-11, lung cancer cell strain A549, colon cancer cell strain HCT116 and other tumor cell strains.

The MTT method utilizes the fact that dehydrogenase related to NADP exists in mitochondria of living cells to reduce exogenous MTT into a difficultly soluble bluish purple crystal (Formazan) and deposits the bluish purple crystal in the cells, but dead cells do not have the function. The purple crystals in the cells were dissolved in dimethyl sulfoxide (DMSO) or a triple solution (10% SDS-5% isobutanol-0.01 mol/L HCl), and the OD value was measured at a wavelength of 570nm using an enzyme-linked immunosorbent assay to indirectly reflect the amount of viable cells.

The specific method comprises the following steps: inoculating the tumor cells to be tested in the logarithmic growth phase of the cells into a 96-well culture plate at a certain cell mass, culturing for 24h, adding the screened sample (directly after the suspension cells are attached to the plate), and making the cells have a CO content of 5% at 37 ℃% ₂ After further culturing for 48 hours under the condition, MTT is added for further culturing for 4 hours, and the crystals are dissolved by DMSO and detected under a microplate reader.

The results of the in vitro antitumor activity of the target compound against the above tumor cell lines at a concentration of 10. Mu.M were as follows (% inhibition):

the pharmacological test result shows that the compound has stronger inhibitory activity on FLT3 and certain selectivity on other kinases, and can be used for preventing or treating clinical diseases related to FLT3, CDK2, CDK4 or CDK6, wherein the diseases can be: leukemia, lymphoma (non-hodgkin lymphoma), hodgkin's disease (also known as hodgkin lymphoma) and myeloma such as Acute Lymphocytic Leukemia (ALL), acute myeloblastic leukemia or Acute Myelogenous Leukemia (AML), acute Promyelocytic Leukemia (APL), chronic Lymphocytic Leukemia (CLL), chronic Myelogenous Leukemia (CML), chronic Neutrophilic Leukemia (CNL), acute undifferentiated cell leukemia (AUL), progression-developed large cell lymphoma (ALCL), adult T-cell ALL, AML with langbeige (triline) myelodysplasia (AML/TMDS), mixed Lineage Leukemia (MLL), myelodysplastic syndrome (ssmd), myelodysplasia (MPD), multiple Myeloma (MM) and myxosarcoma, lung cancer, melanoma, liver cancer, kidney cancer, leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, hyperplastic syndrome, gall bladder cancer, peripheral bladder cancer, melanoma, thyroid cancer, peripheral nerve cell lymphoma, or non-dependent neuroblastoma, e.g., central nervous system lymphoma, peripheral nerve cell lymphoma, neuroblastoma, and non-dependent melanoma (schwanoma).

Detailed Description

The melting point is measured by a b-shaped melting point tube, the medium is methyl silicone oil, and a thermometer is not corrected; ¹ HNMR was performed using JEOL FX90Q type Fourier transform nuclear magnetic resonance apparatus, BRUKERACF-300 type nuclear magnetic resonance apparatus (TMS internal standard); MS is measured by a Nicolet 2000 type Fourier transform mass spectrometer and an MAT-212 type mass spectrometer; the microwave reaction is carried out by using a CEM Discover single-mode microwave instrument.

Example 1

1-methyl-4- (4-nitrobenzyl) piperazine (I-a)

A500 mL single-necked flask was charged with 10g (46.3 mmol) of p-nitrobenzyl bromide and 100mL of methylene chloride, and a mixture of 4.7g (47.0 mmol) of N-methylpiperazine and 7.1g (70.3 mmol) of triethylamine in 20mL of methylene chloride was slowly added dropwise under an ice-water bath (0-5 ℃ C.), followed by heatingReflux 1h, tlc detected disappearance of starting material (ethyl acetate: petroleum ether = 1: 2). 150mL of chloroform and 100mL of a saturated sodium bicarbonate solution were added to the reaction solution, and the mixture was vigorously stirred at room temperature for 30min. The reaction mixture was extracted with chloroform (100 mL. Times.3), and the organic layers were combined and washed once with water and once with saturated sodium chloride (100 mL. Times.1). The anhydrous magnesium sulfate is dried, filtered, and the solvent is evaporated under reduced pressure to obtain 8.5g of light yellow solid with yield of 78.1 percent, and the product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ8.1(d，J＝8.6Hz，2H，ArH)，7.5(d，J＝8.6Hz，2H，ArH)，3.5(s，2H，-CH ₂ -)，2.3-2.5(br，8H，-CH ₂ -×4)，2.15(s，3H，-CH ₃ ).

Example 2

4- ((4-Methylpiperazin-1-yl) methyl) aniline (I-b)

A500 mL single-necked flask was charged with 8.5g (36.2 mmol) of crude I-a, 2.0g of FeO (OH)/C catalyst and 100mL of 95% ethanol, heated under reflux, and a mixture of 25mL of hydrazine hydrate and 20mL of 95% ethanol was slowly added dropwise to remove the starting material by TLC (methanol: chloroform = 1: 15). The mixture is filtered while the mixture is hot, the filter cake is washed twice with hot ethanol (30 mL multiplied by 2), the solvent is distilled off under reduced pressure to obtain a white solid, and the white solid is dried in vacuum to obtain 6.7g of (I-b), wherein the yield is 90.3%. The product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ8.1(d，J＝8.5Hz，2H，ArH)，7.5(d，J＝8.5Hz，2H，ArH)，4.0(s，2H，-NH ₂ )，3.5(s，2H，-CH ₂ -)，2.3-2.5(br，8H，-CH ₂ -×4)，2.1(s，3H，-CH ₃ )

Example 3

N- (4- ((4-methylpiperazin-1-yl) methyl) phenyl-4-nitro-1H-pyrazole-3-carboxamide (I-c)

A250 mL round-bottomed flask was charged with 7.5g (36.6 mmol) of crude I-a', 6.3g (40.1 mmol) of 4-nitro-1H-pyrazole-3-carboxylic acid, 8.4g (44.0 mmol) of EDC. HCl, 6.0g (44.4 mmol) of HOBt, and 100mL of anhydrous DMF, and stirred at room temperature for 24H. Disappearance of the starting material was detected by TLC (methanol: chloroform = 1: 10). Adding the reaction solution into ice water 200mL, precipitating a large amount of light yellow solid, standing, filtering to obtain yellow solid, recrystallizing the obtained crude product with ethyl acetate and methanol mixed solvent to obtain (I-e) 11.1g, with yield of 88.2％。MS[M+H] ⁺ 345.3。 ¹ H NMR(300MHz，DMSO)δ14.2(s，1H，-NH-，Pyrazole)，10.6(s，1H，-NHCO-)，8.8(s，1H，ArH)，7.6(d，J＝8.7Hz，2H，ArH)，7.3(d，J＝8.7Hz，2H，ArH)，3.4(s，2H，-CH ₂ -)，2.3-2.4(br，8H，-CH ₂ -×4)，2.2(s，3H，-CH ₃ ).

Example 4

N- (4- ((4-Methylpiperazin-1-yl) methyl) phenyl-4-amino-1H-pyrazole-3-carboxamide (I-d)

A250 mL single-necked flask was charged with I-C6.0 g (17.4 mmol), feO (OH)/C catalyst 2g and 95% ethanol 100mL, heated to reflux, and slowly added dropwise with a mixture of hydrazine hydrate 25mL and 95% ethanol 20mL, followed by TLC detection of disappearance of starting material (methanol: chloroform = 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30 mL × 2), distilling under reduced pressure to remove solvent to obtain white solid, recrystallizing the crude product with mixed solvent of ethyl acetate and methanol to obtain (I-M') 3.5g with yield of 64%, and MS [ M + H ]] ⁺ 315.8。 ¹ H NMR(300MHz，DMSO)δ12.7(s，1H，Pyrazole)，9.7(s，1H，-NHCO-)，7.7(d，J＝8.6Hz，2H，ArH)，7.1-7.2(m，3H，ArH)，4.7(s，2H，-NH ₂ )，3.3(s，2H，-CH ₂ -)，2.3-2.5(m，8H，-CH ₂ -×4)，2.1(s，3H，-CH ₃ ).

Example 5

4- (4-nitrobenzyl) morpholine (I-e)

10g (46.3 mmol) of p-nitrobenzyl bromide and 100mL of dichloromethane are added to a 500mL single-neck flask, a mixture of 4.1g (47.0 mmol) of morpholine and 7.1g (70.3 mmol) of triethylamine in 20mL of dichloromethane is slowly added dropwise under an ice-water bath (0-5 ℃), after the addition, the mixture is heated under reflux for 1h, and the disappearance of the raw materials is detected by TLC (ethyl acetate: petroleum ether = 1: 2). 150mL of chloroform and 100mL of a saturated sodium bicarbonate solution were added to the reaction solution, and the mixture was vigorously stirred at room temperature for 30min. The reaction mixture was extracted with chloroform (100 mL. Times.3), and the organic layers were combined and washed once with water and once with saturated sodium chloride (100 mL. Times.1). The anhydrous magnesium sulfate is dried, filtered, and the solvent is evaporated under reduced pressure to obtain 8.5g of light yellow solid with yield of 78.1 percent, and the product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ7.6(2H，d，J＝8.7Hz，ArH)，6.9(2H，d，J＝8.7Hz，ArH)，3.5(2H，s，-CH ₂ -)，3.2(4H，m，-OCH ₂ -×2)，2.3(4H，m，-NCH ₂ -×2).

Example 6

4- (Morpholinomethyl) aniline (I-f)

A500 mL single-necked flask was charged with 8.5g (36.2 mmol) of crude I-e, 2.0g of FeO (OH)/C catalyst and 100mL of 95% ethanol, heated to reflux, and a mixture of 25mL of hydrazine hydrate and 20mL of 95% ethanol was slowly added dropwise to remove the starting material by TLC (methanol: chloroform = 1: 15). The solution is filtered while the solution is hot, the filter cake is washed twice with hot ethanol (30 mL multiplied by 2), the solvent is distilled off under reduced pressure to obtain white solid, and the white solid is dried in vacuum to obtain I-f6.5g, wherein the yield is 93.5 percent. The product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ7.6(2H，d，J＝8.5Hz，ArH)，6.9(2H，d，J＝8.5Hz，ArH)，4.9(2H，s，-NH ₂ -)，3.5(2H，s，-CH ₂ -)，3.2(4H，m，-OCH ₂ -×2)，2.3(4H，m，-NCH ₂ -×2).

Example 7

N- (4- (morpholinomethyl) phenyl) -4-nitro-1H-pyrazole-3-carboxamide (I-g)

A250 mL round-bottom flask was charged with 7.0g (36.6 mmol) of crude I-f, 6.3g (40.1 mmol) of 4-nitro-1H-pyrazole-3-carboxylic acid, 8.4g (44.0 mmol) of EDC. HCl, 6.0g (44.4 mmol) of HOBt, and 100mL of anhydrous DMF, and stirred at room temperature for 24H. TLC detection of disappearance of starting material (methanol: chloroform = 1: 10). Adding the reaction solution into ice water 200mL, precipitating a large amount of light yellow solid, standing, filtering to obtain yellow solid, recrystallizing the obtained crude product with mixed solvent of ethyl acetate and methanol to obtain (I-g) 8.6g, and obtaining the yield of 71.0%. MS [ M + H ]] ⁺ 332.3。 ¹ H NMR(300MHz，DMSO)δ14.2(1H，s，-NH-，Pyrazole)，10.7(1H，s，-NHCO-)，8.8(1H，s，ArH)，7.6(2H，d，J＝8.5Hz，ArH)，7.3(2H，d，J＝8.5Hz，ArH)，3.6(4H，m，-OCH ₂ -×2)，3.4(2H，s，-CH ₂ -)，2.4(4H，m，-NCH ₂ -×2).

Example 8

4-amino-N- (4- (morpholinomethyl) phenyl) -1H-pyrazole-3-carboxamide (I-H)

In a 250mL single-necked bottle5.8g (17.4 mmol) of I-g, 2g of FeO (OH)/C catalyst and 100mL of 95% ethanol were added, the mixture was heated under reflux, 25mL of hydrazine hydrate and 20mL of 95% ethanol were slowly added dropwise, and the disappearance of the starting material was detected by TLC (methanol: chloroform = 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30 mL × 2), distilling under reduced pressure to remove solvent to obtain white solid, recrystallizing the crude product with mixed solvent of ethyl acetate and methanol to obtain (I-H) 4.0g, yield 76.0%, and MS [ M + H ]] ⁺ 302.3。 ¹ H NMR(300MHz，DMSO)δ12.7(1H，s，-NH-，Pyrazole)，9.7(1H，s，-NHCO-)，8.8(1H，s，ArH)，7.6(2H，d，J＝8.5Hz，ArH)，7.3(2H，d，J＝8.5Hz，ArH)，4.7(2H，s，-NH ₂ )，3.6(4H，m，-OCH ₂ -×2)，3.3(2H，s，-CH ₂ -)，2.4(4H，m，-NCH ₂ -×2).

Example 9

4- (4-Nitrophenyl) piperazine-1-carboxylic acid tert-butyl ester (I-I)

10g (70.9 mmol) of p-fluoronitrobenzene and 14.6g (106.4 mmol) of potassium carbonate were added to a 500mL single-necked flask, dissolved in 100mL of DMSO, and a solution of N-Boc-piperazine (10.6 g (106.4 mmol) in DMSO was slowly added dropwise under an ice-water bath (0-5 ℃ C.), heated under reflux for 6h after the addition was completed, and the disappearance of the starting material was detected by TLC (ethyl acetate: petroleum ether = 1: 2). 1000ml of water is added to separate out yellow solid, the yellow solid is filtered and dried under reduced pressure to obtain 15.2g of light yellow solid with the yield of 70.1 percent, and the product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ8.14(d，J＝8.7Hz，2H，ArH)，7.85(d，J＝8.7Hz，2H，ArH)，3.45(s，4H，-CH ₂ -×2)，3.39(s，4H，-CH ₂ -×2)，1.35(s，9H，-CH ₃ ×3)。

Example 10

4- (4-aminophenyl) piperazine-1-carboxylic acid tert-butyl ester (I-j)

A500 mL single-necked flask was charged with 10g of crude I-I (32.6 mmol), 1.0g of FeO (OH)/C catalyst and 100mL of 95% ethanol, heated under reflux, and slowly added dropwise with a mixture of 10mL of hydrazine hydrate and 20mL of 95% ethanol to detect the disappearance of the starting material by TLC (methanol: chloroform = 1: 15). The mixture is filtered while hot, the filter cake is washed twice with hot ethanol (30 mL multiplied by 2), the solvent is distilled off under reduced pressure to obtain a white solid, and the white solid is dried in vacuum to obtain 7.86g of (I-j), and the yield is 87.1%. Product has noFurther purification is needed, and the product is directly put into the next reaction. ¹ H NMR(300MHz，DMSO)δ6.94(d，J＝8.7Hz，2H，ArH)，6.22(s，2H，-NH ₂ )，6.15(d，2H，J＝8.7Hz，ArH)，3.45(t，4H，-CH ₂ -×2)，3.39(t，4H，-CH ₂ -×2)，1.35(s，9H，-CH ₃ ×3)，

Example 11

4- (4- (4-Nitro-1H-pyrazole-3-carboxamido) phenyl) piperazine-1-carboxylic acid tert-butyl ester (I-k)

A250 mL round-bottom flask was charged with 5g (18.1 mmol) of crude I-j, 3.1g (19.9 mmol) of 4-nitro-1H-pyrazole-3-carboxylic acid, 4.1g (21.7 mmol) of EDC. HCl, 2.9g (21.7 mmol) of HOBt and 50mL of anhydrous DMF, and stirred at room temperature for 24H. TLC detection of disappearance of starting material (methanol: chloroform = 1: 10). Adding the reaction solution into 200mL of ice water, precipitating a large amount of light yellow solid, standing, filtering to obtain a yellow solid, and recrystallizing the obtained crude product with a mixed solvent of ethyl acetate and methanol to obtain 4.7g of (I-k), wherein the yield is 62.4%. MS [ M + H ]] ⁺ 417.2. ¹ H NMR(300MHz，DMSO)δ11.46(s，1H，-NH-，Pyrazole)，9.53(s，1H，-NHCO-)，7.88(s，1H，ArH)，7.84(d，J＝8.7Hz，2H，ArH)，7.45(d，J＝8.7Hz，2H，ArH)，3.45(t，4H，-CH ₂ -×2)，3.39(t，4H，-CH ₂ -×2)，1.45(s，9H，-CH ₃ ×3)。

Example 12

4- (4- (4-amino-1H-pyrazole-3-carboxamido) phenyl) piperazine-1-carboxylic acid tert-butyl ester (I-l)

A250 mL single-necked flask was charged with I-k 5.0g (12.0 mmol), feO (OH)/C catalyst 0.5g, and 95% ethanol 60mL, heated under reflux, hydrazine hydrate 5mL was slowly added dropwise, and the starting material disappeared by TLC detection (methanol: chloroform = 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30 mL × 2), distilling under reduced pressure to remove solvent to obtain white solid, recrystallizing the crude product with mixed solvent of ethyl acetate and methanol to obtain (I-1) 3.9g, yield 85.3%, and MS [ M + H ]] ⁺ 386.2. ¹ H NMR(300MHz，DMSO)δ11.56(s，1H，-NH-，Pyrazole)，9.53(s，1H，-NHCO-)，7.88(s，1H，ArH)，7.84(d，2H，J＝8.7Hz，ArH)，7.44(s，2H，-NH ₂ )，6.45(d，J＝8.7Hz，2H，ArH)，3.55(t，4H，-CH2-×2)，3.49(t，4H，-CH ₂ -×2)，1.45(s，9H，-CH ₃ ×3)。

Example 13

Morpholino (4-nitrophenyl) methanone (I-m)

In a 500mL single neck flask were added 10g (46.3 mmol) of p-nitrobenzoic acid, 4.1g (50.6 mmol) of morpholine, EDCI (50.6 mmol) and HOBT (69 mmol), 100mL of dichloromethane was added, stirring was carried out at ambient temperature for 5h, and disappearance of starting material was detected by TLC (ethyl acetate: petroleum ether = 1: 2). DCM was spin-dried under reduced pressure, water was added, extraction was performed with chloroform (100 mL. Times.3), and the organic layers were combined, washed once with water and once with saturated sodium chloride (100 mL. Times.1), respectively. The anhydrous magnesium sulfate is dried, filtered, and the solvent is removed by evaporation under reduced pressure to obtain 8.5g of light yellow solid, the yield is 88.1%, and the product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ7.7(2H，d，J＝8.7Hz，ArH)，7.0(2H，d，J＝8.7Hz，ArH)，3.7(4H，m，-OCH ₂ -×2)，3.1(4H，m，-NCH ₂ -×2).

Example 14

(4-aminophenyl) (morpholino) methanone (I-n)

A500 mL single-necked flask was charged with 8.5g (36.2 mmol) of crude I-m, 2.0g of FeO (OH)/C catalyst and 100mL of 95% ethanol, heated under reflux, and a mixture of 25mL of hydrazine hydrate and 20mL of 95% ethanol was slowly added dropwise to remove the starting material by TLC (methanol: chloroform = 1: 15). The mixture is filtered while the mixture is hot, the filter cake is washed twice with hot ethanol (30 mL multiplied by 2), the solvent is removed by evaporation under reduced pressure to obtain a white solid, and the white solid is dried in vacuum to obtain I-n 6.5g, wherein the yield is 93.5 percent. The product is directly put into the next reaction without further purification. ¹ H NMR(300MHz，DMSO)δ7.5(2H，d，J＝8.5Hz，ArH)，6.8(2H，d，J＝8.5Hz，ArH)，5.2(2H，s，-NH ₂ )，3.6(4H，m，-OCH ₂ -×2)，3.0(4H，m，-NCH ₂ -×2).

Example 15

N- (4- (morpholine-4-carbonyl) phenyl) -4-nitro-1H-pyrazole-3-carboxamide (I-o)

A250 mL round-bottomed flask was charged with 7.0g (36.6 mmol) of crude I-n, 6.3g (40.1 mmol) of 4-nitro-1H-pyrazole-3-carboxylic acid, 8.4g (44.0 mmol) of EDC. HCl, 6.0g (44.4 mmol) of HOBt, and 100mL of anhydrous DMF, and stirred at room temperature for 24 hours. TLC detection raw materialDisappearance (methanol: chloroform = 1: 10). Adding the reaction solution into 200mL of ice water, precipitating a large amount of light yellow solid, standing, filtering to obtain a yellow solid, and recrystallizing the obtained crude product with a mixed solvent of ethyl acetate and methanol to obtain 8.6g of (I-o), wherein the yield is 72.0%. MS [ M + H ]] ⁺ 346.1。 ¹ H NMR(300MHz，DMSO)δ12.7(1H，s，-NH-，Pyrazole)，9.6(1H，s，-NHCO-)，8.4(1H，s，ArH)，7.7(2H，d，J＝8.5Hz，ArH)，7.1(2H，d，J＝8.5Hz，ArH)，3.5(4H，m，-OCH ₂ -×2)，2.6(4H，m，-NCH ₂ -×2).

Example 16

4-amino-N- (4- (morpholine-4-carbonyl) phenyl) -1H-pyrazole-3-carboxamide (I-p)

A250 mL single-necked flask was charged with I-05.8g (17.4 mmol), feO (OH)/C catalyst 2g and 95% ethanol 100mL, heated to reflux, and slowly added dropwise with a mixture of hydrazine hydrate 25mL and 95% ethanol 20mL, followed by TLC detection of disappearance of starting material (methanol: chloroform = 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30 mL × 2), distilling under reduced pressure to remove solvent to obtain white solid, recrystallizing the crude product with mixed solvent of ethyl acetate and methanol to obtain (I-P) 4.0g, yield 76.0%, and MS [ M + H ]] ⁺ 316.3。 ¹ H NMR(300MHz，DMSO)δ12.6(1H，s，-NH-，Pyrazole)，9.4(1H，s，-NHCO-)，8.3(1H，s，ArH)，7.5(2H，d，J＝8.5Hz，ArH)，7.2(2H，d，J＝8.5Hz，ArH)，4.8(2H，s，-NH ₂ )，3.7(4H，m，-OCH ₂ -×2)，2.9(4H，m，-NCH ₂ -×2).

Example 17

In a 50mL single-necked flask were added I-1150mg (0.39 mmol), 4-chloro-6, 7-dihydro-5H-cyclopenta [ d ]]Heating and refluxing pyrimidine 72.4mg (0.47 mmol) and 50% acetic acid aqueous solution 5mL, detecting disappearance of raw material by TLC (methanol: chloroform = 1: 10), cooling reaction solution to room temperature, adjusting pH to 8-9 with saturated sodium bicarbonate aqueous solution, precipitating solid, suction-filtering, extracting filtrate with ethyl acetate 3 times (50 mL. Times.3), combining extracts, drying with anhydrous magnesium sulfate, vacuum-filtering, evaporating off solvent under reduced pressure, combining with filter cake, and using di (methylene) after combining with filter cakeDissolving chloromethane, adding trifluoroacetic acid with the same volume, and stirring at normal temperature for 3 hours. Evaporating dichloromethane under reduced pressure, adding water to adjust pH to 8-9, vacuum filtering to obtain solid, subjecting the crude product to column chromatography (methanol: chloroform = 1: 15) to obtain (I-1) 138mg, yield 87%, [ M + H ]] ⁺ 405.2. ¹ H NMR(300MHz，DMSO)δ13.40(s，1H，pyrazole)，10.10(s，1H，-NH-)，9.11(s，1H，-NHCO-)，8.49(s，1H，ArH)，7.67(d，J＝7.3Hz，2H，ArH)，6.94(d，J＝7.3Hz，2H，ArH)，7.52(d，J＝3.5Hz，3H，ArH)，3.14(m，4H)，2.75-2.81(m，4H)，2.06-2.09(s，2H，-CH ₂ -).

Example 18

In a similar manner to the preparation of I-1, a white solid (I-2) was obtained in 53% yield and a white solid was obtained in 71% yield. MS [ M + H ]] ⁺ 460.3. ¹ H NMR(300MHz，DMSO)δ13.49(s，1H，pyrazole)，10.11(s，1H，-NH-)，9.27(s，1H，-NHCO-)，8.63(s，1H，ArH)，7.71(d，J＝8.2Hz，2H，ArH)，6.97(d，J＝8.2Hz，2H，ArH)，3.33(s，4H，-CH ₂ -×2)，3.26(s，4H，-CH ₂ -×2)，2.60(m，2H，-CH ₂ -)，2.52(s，2H，-CH ₂ -)，1.83(m，2H，-CH ₂ -)，1.77(m，2H，-CH ₂ -).

Example 19

In a 50mL single-necked flask were added I-H117 mg (0.39 mmol), 4-chloro-6, 7-dihydro-5H-cyclopenta [ d ]]Heating and refluxing 72.4mg (0.47 mmol) of pyrimidine and 5mL of 50% acetic acid aqueous solution, detecting disappearance of raw materials by TLC (methanol: chloroform = 1: 10), cooling reaction liquid to room temperature, adjusting pH to 8-9 with saturated sodium bicarbonate aqueous solution, precipitating solid, carrying out suction filtration, extracting filtrate for 3 times (50 mL × 3) with ethyl acetate, combining extract liquor, drying with anhydrous magnesium sulfate, carrying out suction filtration, carrying out reduced pressure evaporation to remove solvent, combining with filter cake, carrying out column chromatography on crude product (methanol: chloroform = 1: 15) to obtain (I-3) 92mg with yield of 0.56%, [ M + H ] (M + H) (M + H) ()] ⁺ 420.2. ¹ H NMR(300MHz，DMSO)δ13.45(s，1H，pyrazole)，10.19(s，1H，-NH-)，9.23(s，1H，-NHCO-)，8.55(s，1H，ArH)，8.44(s，1H，ArH)，7.73(d，J＝8.6Hz，2H，ArH)，6.96(d，J＝8.6Hz，2H，ArH)，3.41(s，2H，-CH ₂ -)，3.33(s，4H，-CH ₂ -×2)，3.11(s，4H，-CH ₂ -×2)，2.75-2.81(m，4H，-CH ₂ -×2)，2.01-2.03(s，2H，-CH ₂ -).

Example 20

In a similar manner to that for the preparation of I-3, white solid (I-4) was obtained in 47% yield and MS [ M + H ]] ⁺ 434.2. ¹ H NMR(300MHz，DMSO)δ13.41(s，1H，pyrazole)，10.04(s，1H，-NH-)，9.33(s，1H，-NHCO-)，9.12(s，1H，ArH)，8.44(s，1H，ArH)，7.73(d，J＝8.8Hz，2H，ArH)，6.97(d，J＝8.8Hz，2H，ArH)，3.42(s，2H，-CH ₂ -)，3.23(s，4H，-CH ₂ -×2)，3.02(s，4H，-CH ₂ -×2)，2.61(m，2H，-CH ₂ -)，2.43(m，2H，-CH ₂ -)，1.82(m，2H，-CH ₂ -)，1.75(m，2H，-CH ₂ -).

Example 21

In a manner similar to that for preparation of I-3, white solid (I-5) was obtained in 54% yield of MS [ M + H ]] ⁺ 434.2. ¹ H NMR(300MHz，DMSO)13.44(s，1H，pyrazole)，10.12(s，1H，-NH-)，9.21(s，1H，-NHCO-)，8.42(s，1H，ArH)，7.76(d，J＝8.2Hz，2H，ArH)，6.99(d，J＝8.2Hz，2H，ArH)，3.40(s，2H，-CH ₂ -)，3.27(s，4H，-CH ₂ -×2)，3.01(s，4H，-CH ₂ -×2)，2.75-2.81(m，4H，-CH ₂ -×2)，2.61(s，3H，-CH ₃ )，2.00-2.04(s，2H，-CH ₂ -).

Example 22

In a similar manner to that for the preparation of I-3, pale yellow solid (I-6) was obtained in 49% yield, MS [ M + H] ⁺ 448.2. ¹ H NMR(300MHz，DMSO)δ13.44(s，1H，pyrazole)，10.12(s，1H，-NH-)，9.29(s，1H，-NHCO-)，9.11(s，1H，ArH)，7.77(d，J＝8.6Hz，2H，ArH)，6.99(d，J＝8.6Hz，2H，ArH)，3.42(s，2H，-CH ₂ -)，3.21(s，4H，-CH ₂ -×2)，3.07(s，4H，-CH ₂ -×2)，2.62(m，3H，-CH ₃ )，2.56(m，4H，-CH ₂ -)，1.88(m，2H，-CH ₂ -)，1.74(m，2H，-CH ₂ -).

Example 23

In a similar manner to preparation of I-1, pale yellow solid (I-7) was obtained in 53% yield of MS [ M + H ]] ⁺ 419.2. ¹ H NMR(300MHz，DMSO)δ13.40(s，1H，pyrazole)，10.10(s，1H，-NH-)，9.11(s，1H，-NHCO-)，8.49(s，1H，ArH)，7.67(d，J＝8.4Hz，2H，ArH)，6.95(d，J＝8.4Hz，2H，ArH)，3.52(m，4H，-CH ₂ -×2)，3.01(s，4H，-CH ₂ -×2)，2.75-2.81(m，4H，-CH ₂ -×2)，2.51(s，3H，-CH ₃ )，2.06-2.07(m，2H，-CH ₂ -).

Example 24

In a similar manner to preparation I-1, pale yellow solid (I-8) was obtained in 51% yield and MS [ M + H ]] ⁺ 433.2. ¹ H NMR(300MHz，DMSO)δ13.48(s，1H，pyrazole)，10.16(s，1H，-NH-)，9.36(s，1H，-NHCO-)，8.50(s，1H，ArH)，7.71(d，J＝8.6Hz，2H，ArH)，6.99(d，J＝8.6Hz，2H，ArH)，3.33-3.35(m，4H，-CH ₂ -×2)，3.21-3.23(s，2H，-CH ₂ -×2)，2.59-2.61(m，2H，-CH ₂ -)，2.46(s，5H，-CH ₃ ，-CH ₂ -)，1.76-1.84(m，4H，-CH ₂ -×2).

Example 25

In a 50mL single-necked flask were added I-d 123mg (0.39 mmol), 4-chloro-2-methyl-6, 7-dihydro-5H-cyclopenta [ d]79.0mg (0.47 mmol) of pyrimidine and 5mL of 50% acetic acid aqueous solution are heated and refluxed, TLC detection raw materials disappear (methanol: chloroform = 1: 10), reaction liquid is cooled to room temperature, the pH is adjusted to 8-9 by saturated sodium bicarbonate aqueous solution, solid is separated out, suction filtration is carried out, filtrate is extracted by ethyl acetate for 3 times (50 mL multiplied by 3), extract liquor is combined, anhydrous magnesium sulfate is dried, the solvent is evaporated under reduced pressure after suction filtration and is combined with filter cakes, crude products are subjected to column chromatography (methanol: chloroform = 1: 15) to obtain (I-9) 91mg, the yield is 52%, [ M + H ], [] ⁺ 447.2. ¹ H NMR(300MHz，DMSO)δ13.44(s，1H，pyrazole)，10.11(s，1H)，9.42(s，1H)，8.61(s，1H，ArH)，7.72(d，J＝8.2Hz，2H，ArH)，6.96(d，J＝8.2Hz，2H，ArH)，3.42(s，2H，-CH ₂ -)，2.62(s，3H，-CH ₃ )，2.35(s，8H，-CH ₂ -×4)，2.76-2.88(m，4H，-CH ₂ -×2)，2.12(s，3H，-CH ₃ )，2.02-2.05(m，2H，-CH ₂ -).

Example 26

In a similar manner to that for the preparation of I-9, a pale yellow solid (I-10) was obtained in 66% yield, MS [ M + H%] ⁺ 461.2. ¹ H NMR(300MHz，DMSO)δ13.43(s，1H，pyrazole)，10.11(s，1H，-NH-)，9.40(s，1H，-NHCO-)，8.46(s，1H，ArH)，7.69(d，J＝8.4Hz，2H，ArH)，6.97(d，J＝8.4Hz，2H，ArH)，3.42(s，2H，-CH ₂ -)，2.60(s，3H，-CH ₃ )，2.56-2.59(m，2H，-CH ₂ -)，2.38(s，10H，-CH ₂ -×5)，2.11(s，3H，-CH ₃ )，1.76-1.84(m，4H，-CH ₂ -×2).

Example 27

In a similar manner to preparation I-1, pale yellow solid (I-11) was obtained in 64% yield and MS [ M + H ]] ⁺ 473.2. ¹ H NMR(300MHz，DMSO)δ13.42(s，1H，pyrazole)，10.09(s，1H，-NH-)，9.21(s，1H，-NHCO-)，8.54(s，1H，ArH)，7.73(d，J＝8.4Hz，2H，ArH)，6.99(d，J＝8.4Hz，2H，ArH)，3.45(m，4H，-CH ₂ -×2)，3.01(s，4H，-CH ₂ -×2)，2.75-2.81(m，4H，-CH ₂ -×2)，2.06-2.07(m，2H，-CH ₂ -).

Example 28

In a similar manner to that for preparation I-1, pale yellow solid (I-12) was obtained in 49% yield and MS [ M + H ]] ⁺ 487.2. ¹ H NMR(300MHz，DMSO)δ13.42(s，1H，pyrazole)，10.14(s，1H，-NH-)，9.33(s，1H，-NHCO-)，8.56(s，1H，ArH)，7.66(d，J＝8.2Hz，2H，ArH)，6.94(d，J＝8.2Hz，2H，ArH)，3.34-3.37(m，4H，-CH ₂ -×2)，3.20-3.23(s，4H，-CH ₂ -×2)，2.59-2.61(m，2H，-CH ₂ -)，2.44(s，2H，-CH ₂ -)，1.74-1.81(m，4H，).

Example 29

In a similar manner to preparation of I-1, pale yellow solid (I-13) was obtained in 67% yield and MS [ M + H ]] ⁺ 447.3. ¹ H NMR(300MHz，DMSO)δ13.41(s，1H，pyrazole)，10.11(s，1H，-NH-)，9.22(s，1H，-NHCO-)，8.51(s，1H，ArH)，7.72(d，J＝8.4Hz，2H，ArH)，6.98(d，J＝8.4Hz，2H，ArH)，3.44(m，4H，-CH ₂ -×2)，3.31(m，1H)，3.04(s，4H，-CH ₂ -×2)，2.77-2.84(m，4H，-CH ₂ -×2)，2.04-2.06(m，2H，-CH ₂ -)，1.34(d，J＝6.8Hz，6H).

Example 30

In a similar manner to preparation of I-1, a pale yellow solid (I-14) was obtained in a yield of 76%, MS [ M + H ]] ⁺ 461.3. ¹ H NMR(300MHz，DMSO)δ13.49(s，1H，pyrazole)，10.11(s，1H，-NH-)，9.30(s，1H，-NHCO-)，8.48(s，1H，ArH)，7.69(d，J＝8.4Hz，2H，ArH)，6.98(d，J＝8.4Hz，2H，ArH)，3.35-3.38(m，4H，-CH ₂ -×2)，3.31(m，1H)，3.18-3.23(s，4H，-CH ₂ -×2)，2.56-2.58(m，2H，-CH ₂ -)，2.41(s，2H，-CH ₂ -)，1.70-1.84(m，4H，-CH ₂ -×2)，1.32(d，J＝6.6Hz，6H).

Example 31

In a similar manner to preparation I-1, pale yellow solid (I-15) was obtained in a yield of 57%, MS [ M + H ]] ⁺ 433.2. ¹ H NMR(300MHz，DMSO)δ13.49(s，1H，pyrazole)，10.41(s，1H，-NH-)，9.23(s，1H，-NHCO-)，8.55(s，1H，ArH)，7.77(d，J＝8.4Hz，2H，ArH)，6.97(d，J＝8.4Hz，2H，ArH)，3.41-3.46(m，4H，-CH ₂ -×2)，3.01-3.06(m，4H，-CH ₂ -×2)，2.90(m，2H，-CH ₂ -)，2.76-2.82(m，4H，-CH ₂ -×2)，2.02-2.04(m，2H，-CH ₂ -)，1.37(t，J＝7.4Hz，3H).

Example 32

In a similar manner to preparation of I-1, a pale yellow solid (I-16) was obtained in a yield of 47%, MS [ M + H ]] ⁺ 447.2 ¹ H NMR(300MHz，DMSO)δ13.39(s，1H，pyrazole)，10.08(s，1H，-NH-)，9.32(s，1H，-NHCO-)，8.45(s，1H，ArH)，7.71(d，J＝8.4Hz，2H，ArH)，6.96(d，J＝8.4Hz，2H，ArH)，3.33-3.35(m，4H，-CH ₂ -×2)，3.11-3.15(s，4H，-CH ₂ -×2)，2.91(m，2H，-CH ₂ -)，2.54-2.56(m，2H，-CH ₂ -)，2.44(s，2H，-CH ₂ -)，1.68-1.78(m，4H，-CH ₂ -×2)，1.36(t，J＝7.2Hz，3H).

Example 33

In a similar manner to preparation I-1, pale yellow solid (I-17) was obtained in 45% yield of MS [ M + H ]] ⁺ 439.2. ¹ H NMR(300MHz，DMSO)δ13.49(s，1H，pyrazole)，10.04(s，1H，-NH-)，9.27(s，1H，-NHCO-)，8.50(s，1H，ArH)，7.68(d，J＝8.4Hz，2H，ArH)，6.96(d，J＝8.4Hz，2H，ArH)，3.42(m，4H，-CH ₂ -×2)，3.08(m，4H，-CH ₂ -×2)，2.71-2.78(m，4H，-CH ₂ -×2)，2.00-2.04(m，2H，-CH ₂ -).

Example 34

In a similar manner to preparation I-1, pale yellow solid (I-18) was obtained in 29% yield, MS [ M + H]+453.2. ¹ H NMR(300MHz，DMSO)δ13.43(s，1H，pyrazole)，10.05(s，1H，-NH-)，9.26(s，1H，-NHCO-)，8.43(s，1H，ArH)，7.70(d，J＝8.4Hz，2H，ArH)，6.98(d，J＝8.4Hz，2H，ArH)，3.33-3.36(m，4H，-CH ₂ -×2)，3.18-3.22(s，4H，-CH ₂ -×2)，2.58-2.61(m，2H，-CH ₂ -)，2.42(s，2H，-CH ₂ -)，1.70-1.78(m，4H，).

Example 35

In a similar manner to preparation I-1, pale yellow solid (I-19) was obtained in 37% yield and MS [ M + H ]]+445.2. ¹ H NMR(300MHz，DMSO)δ13.41(s，1H，pyrazole)，10.02(s，1H，-NH-)，9.21(s，1H，-NHCO-)，8.42(s，1H，ArH)，7.68(d，J＝8.4Hz，2H，ArH)，6.96(d，J＝8.4Hz，2H，ArH)，3.32-3.34(m，4H，-CH ₂ -×2)，3.16-3.23(s，4H，-CH ₂ -×2)，2.56-2.60(m，2H，-CH ₂ -)，2.42(s，2H，-CH ₂ -)，2.21(m，1H)，1.68-1.74(m，4H，)，1.07(m，4H).

Example 36

In a similar manner to preparation of I-1, a pale yellow solid (I-20) was obtained in 34% yield and MS [ M + H ]] ⁺ 459.3. ¹ H NMR(300MHz，DMSO)δ13.49(s，1H，pyrazole)，10.12(s，1H，-NH-)，9.19(s，1H，-NHCO-)，8.48(s，1H，ArH)，7.71(d，J＝8.4Hz，2H，ArH)，6.94(d，J＝8.4Hz，2H，ArH)，3.35-3.39(m，4H，-CH ₂ -×2)，3.18-3.22(s，4H，-CH ₂ -×2)，2.58-2.61(m，2H，-CH ₂ -)，2.42(s，2H，-CH ₂ -)，2.24(m，1H)，1.74-1.82(m，4H，)，1.11(m，4H).

Example 37

In a similar manner to preparation I-1, pale yellow solid (I-21) was obtained in 42% yield, MS [ M + H] ⁺ 446.5. ¹ H NMR(300MHz，DMSO)δ13.47(s，1H，pyrazole)，10.06(s，1H，-NH-)，9.12(s，1H，-NHCO-)，8.52(s，1H，ArH)，7.74(d，J＝8.4Hz，2H，ArH)，6.98(d，J＝8.4Hz，2H，ArH)，3.53(m，4H，-CH ₂ -×2)，3.01(s，4H，-CH ₂ -×2)，2.74-2.82(m，4H，-CH ₂ -×2)，2.49(s，3H，-CH ₃ )，2.06-2.15(m，6H，-CH ₂ -×3).

Example 38

4- ((6, 7-dihydro-5H-cyclopenta [ d ] pyrimidin-4-yl) amino) -N- (4- (morpholine-4-carbonyl) phenyl) -1H-pyrazole-3-carboxamide (I-22)

In a 50mL single necked flask was added I-q 117mg (0.39 mmol), 4-chloro-6, 7-dihydro-5H-cyclopenta [ d ]]Heating and refluxing 72.4mg (0.47 mmol) of pyrimidine and 5mL of 50% acetic acid aqueous solution, detecting by TLC that raw materials disappear (methanol: chloroform = 1: 10), cooling reaction liquid to room temperature, adjusting pH to 8-9 with saturated sodium bicarbonate aqueous solution, precipitating solid, carrying out suction filtration, extracting filtrate for 3 times (50 mL × 3) with ethyl acetate, combining extract liquor, drying with anhydrous magnesium sulfate, carrying out suction filtration, carrying out reduced pressure evaporation to remove solvent, combining with filter cake, carrying out column chromatography on crude product (methanol: chloroform = 1: 15) to obtain (I-22) 84mg with yield of 48%, [ M + H ] (M + H) ]] ⁺ 434.2. ¹ H NMR(300MHz，DMSO)δ13.44(s，1H，pyrazole)，10.10(s，1H，-NH-)，9.24(s，1H，-NHCO-)，8.53(s，1H，ArH)，8.41(s，1H，ArH)，7.70(d，J＝8.6Hz，2H，ArH)，6.98(d，J＝8.6Hz，2H，ArH)，3.39(s，4H，-CH ₂ -×2)，3.10(s，4H，-CH ₂ -×2)，2.76-2.89(m，4H，-CH ₂ -×2)，1.99-2.02(s，2H，-CH ₂ -).

Drawings

FIG. 1 is a diagram of the general formula (I).

Claims

1. A compound of the general formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is ¹ Is hydrogen, halogenated straight or branched chain saturated hydrocarbon group having 1 to 8 carbon atoms, cyclopropyl, cyclobutyl or halogen;

x and Y represent N atoms;

A ¹ is NH; a. The ² Is a bond or methylene;

Q ¹ represents a phenyl group;

Q ² is piperazinyl, morpholinyl, tetrahydropyrrolyl, piperidinyl, homopiperazinyl, pyranyl, tetrahydrofuranyl, N-methylpiperidin-4-yl, N-methylpiperazinyl or 3-methylpiperidin-1-yl;

the value range of n is 1-4.

2. The compound of claim 1 having a structure selected from the group consisting of:

4- ((2-methyl-6,7,8,9-tetrahydro-5H-cyclohepta [ d ] pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-21).

3. A pharmaceutically acceptable salt of a compound according to any one of claims 1-2, wherein the pharmaceutically acceptable salt is selected from the group consisting of acid addition salts of the above compounds with: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, or mandelic acid.

4. A pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a clinical condition associated with FLT 3.

6. The use according to claim 5, wherein the disease associated with FLT3 is lung cancer, melanoma, liver cancer, kidney cancer, leukemia, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gall bladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, gastrointestinal cancer, astrocytoma, neuroblastoma, glioma, schwannoma, mesothelioma, non-insulin dependent diabetes mellitus, autoimmune disease.