CN107245073B

CN107245073B - 4- (aromatic heterocycle substituted) amino-1H-3-pyrazolecarboxamide FLT3 inhibitor and application thereof

Info

Publication number: CN107245073B
Application number: CN201710567840.XA
Authority: CN
Inventors: 卢帅; 王越; 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: 2017-07-11
Filing date: 2017-07-11
Publication date: 2020-04-17
Anticipated expiration: 2037-07-11
Also published as: CN107245073A

Abstract

The invention relates to a novel 4- (aromatic heterocyclic substituted) amino-1H-3-pyrazole formamide compound or pharmaceutically acceptable salt, solvate, isomer, ester, acid, metabolite or prodrug thereof, a preparation method thereof, a medicinal composition containing the compound and medical application thereof.

Description

4- (aromatic heterocycle 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

Protein kinases are enzymatic components of signal transduction pathways that catalyze the transfer of the terminal phosphate of ATP to the hydroxyl group of tyrosine, serine, and/or threonine residues of proteins. Overexpression or inappropriate expression of normal or mutant protein kinases in mammals has been the subject of extensive research and has been shown to play a significant role in the development of a number of diseases, including diabetes, angiogenesis, psoriasis, restenosis, ocular diseases, schizophrenia, rheumatoid arthritis, atherosclerosis, cardiovascular diseases and cancer. Inhibitors of protein kinases therefore have particular application in the treatment of human and animal diseases.

In recent years, protein kinase inhibitors have been a hot spot for research and development of antitumor drugs, 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(Fms-like tyrosine Kinase, FLT3), also known as Fetal Liver Kinase-2 (Fetal Liver Kinase-2, FLK2) and Stem Cell Kinase-1 (Stem Cell Kinase-1, STK1), are a third class of receptor tyrosine kinases. FLT3 gene, located at chromosome 13q12, is an early hematopoietic growth factor receptor gene discovered in 1991, and its encoded FLT3 receptor belongs to a member of tyrosine kinase receptor family. Upon binding of the extracellular domain of FLT3 receptor to its endogenous ligand, FLT3 forms a homo-or heterodimeric complex, causing its tyrosine kinase activity to be activated, the activation loop to open, promoting the attachment of the substrate protein to the ATP binding site, further catalyzing the phosphorylation of the substrate protein, thereby mediating a series of downstream signal transduction, leading to the proliferation and differentiation of cells. FLT3 receptor is widely distributed in various tissues such as bone marrow hematopoietic stem/progenitor cells, thymus, lymph, placenta, brain, gonads, etc. However, the FLT3 gene mutation (mainly including the internal tandem repeat mutation of the juxtamembrane domain and the point mutation of the tyrosine kinase domain) and over-expression can cause various hematological malignant diseases such as acute myelogenous leukemia. Thus, targeting FLT3 for tumor therapy has been a focus of research.

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, in hematological malignancies FLT3 expression at high levels or FLT3 mutations cause uncontrolled FLT3 receptors and downstream molecular channels to induce activation of the likely RAS, JAK, PI3K, etc. 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 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. Currently, targeted inhibition of FLT3 and mutant FLT3 becomes a research hotspot, mainly aiming at developing small-molecule tyrosine kinase inhibitors, and inhibiting the activity of the inhibitors by competing with the FLT3 tyrosine kinase for ATP binding sites. Kinase inhibitors that inhibit FLT3 that have been clinically developed 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 effective doses for inhibiting FLT3 in AML patients over the range of drug-tolerance levels. 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, the FLT3 mutation was the 1 st mutation 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, only the FLT3 inhibitor PKC412 developed by nova was marketed for oral treatment of adult patients with newly diagnosed acute myeloid leukemia with specific genetic mutation FLT 3. 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 of the invention has excellent FLT3 selectivity and inhibition activity. Has strong inhibitory activity to various tumor cell strains 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 the general formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is¹Represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, heterocycloalkyl, alkylamino, alkoxyalkyl, alkoxyformyl, alkylcarbamoyl, aralkyl, diarylalkyl, arylheterocycloalkyl, aryl or heteroaromatic;

R²represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, heterocycloalkyl, alkylamino, alkoxyalkyl, alkoxyformyl, alkylcarbamoyl, aralkyl, aryloxy, arylamino, arylthio, aralkyloxy, aralkylamino, aralkylthio, diarylalkyl, arylheterocycloalkyl, aryl or arylheterocycle;

x, Y, Z each independently represents an N atom or a CH group; wherein the CH radical may optionally be substituted by R³Substituted, R³Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

A¹each independently represents NH, O, S or an alkylene radical; wherein each NH radical or alkylene radical independently may be optionally substituted by R⁴Substituted, R⁴Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

A²each 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 with R⁵Substituted, R⁵Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

Q¹is selected from aryl or aromatic heterocycle, wherein aryl or aromatic heterocycle independently of each other may be optionally substituted by one or more R⁶Substituted, R⁶Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

Q²is selected from aryl, aromatic heterocycle, C₃-C₈Aliphatic carbocycle, heterocycloalkyl, -NH of₂or-OH, wherein aryl, aromatic heterocycle, C₃-C₈Each independently of the other, may be optionally substituted with one or more R⁷Substituted, R⁷Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic; -NH₂or-OH independently of one another may optionally be substituted by one or more R⁸Substituted, R⁸Can be hydrogen, alkyl, haloalkyl, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic;

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 losing one hydrogen atom to a cyclic saturated hydrocarbon group having 3 to 8 carbon atoms to which a straight or branched 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 (wherein m is an integer of 0 to 2), the remaining ring atoms being 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 bonded; 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 or pyridazinyl; or a bicyclic heterocycle selected from quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2, 3-dihydrobenzo [1, 4] dioxinyl or 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:

wherein R is¹Represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkylamino, heterocycloalkyl, alkoxyalkyl, alkoxyformyl or alkylcarbamoyl;

x, Y, Z each independently represents an N atom or a CH group; wherein the CH radical may optionally be substituted by R²Substituted, R³Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

A¹each independently represents NH, O, S or an alkylene radical; wherein each NH radical or alkylene radical independently may be optionally substituted by R⁴Substituted, R⁴Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

A²each 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 R⁵Substituted, R⁵Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

Q¹is selected from aryl or aromatic heterocycle, wherein aryl or aromatic heterocycle independently of each other may be optionally substituted by one or more R⁶Substituted, R⁷Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio or alkoxyalkyl;

Q²is selected from aryl, aromatic heterocycle, C₃-C₈Aliphatic carbocycle, heterocycloalkyl, -NH of₂or-OH, wherein aryl, aromatic heterocycle, C₃-C₈Each independently of the other, may be optionally substituted with one or more R⁷Substituted, R⁷Can be hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic; -NH₂or-OH independently of one another may optionally be substituted by one or more R⁸Substituted, R⁸May be hydrogen, alkyl, haloalkyl, alkoxyalkyl, aralkyl, diarylalkyl, aryl, or heteroaromatic.

Another preferred embodiment of the present invention is:

wherein R is¹Represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxyl, mercapto, alkoxy, alkylthio or alkylamino;

R²represents hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxy, mercapto, alkoxy, alkylthio, heterocycloalkyl, alkylamino, alkoxyalkyl, alkoxyformyl, mercapto, or a salt thereof,An alkylcarbamoyl group, an aralkyl group, an aryloxy group, an arylamino group, an arylthio group, an aralkyloxy group, an aralkylamino group, an aralkylthio group, a diarylalkyl group, an arylheterocycloalkyl group, an aryl group, or an arylheterocycle;

x, Y, Z each independently represents an N atom or a CH group;

A¹each independently represents NH, O, S or an alkylene radical;

A²each 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;

Another preferred embodiment of the present invention is:

wherein R is¹Represents hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyano or halogen;

wherein R is²An alkyl group selected from: cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, or a heterocycloalkyl group of: oxetanyl, oxacyclopentyl, oxacyclohexenylAlkyl, tetrahydropyrrolyl, piperidinyl, N-methylpiperidin-4-yl, morpholinyl, N-methylpiperazinyl, 3-methylpiperidin-1-yl, piperazinyl, or alkoxy and alkylamino of the following: cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, cyclopentylmethoxy, cyclopentylethoxy, cyclopropylamino, cyclobutylamino, cyclopentylamino, cyclohexylamino, cycloheptylamino, or the following aryl and heteroaryl rings: phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, pyrrolyl, furanyl, thienyl, pyridyl, pyrimidinyl, or aryloxy and arylamino groups of: phenoxy, naphthoxy, anilino, naphthylamino, or the following aralkyloxy and aralkylamino groups: benzyloxy, benzylamino;

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

A¹represents NH;

A²each independently represents a bond, alkylene, or C (O);

Q¹is 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;

Q²is 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 a substituted amino group, a substituted oxy group selected from: n, N-dipropylamino, N-diethylamino, N-dimethylamino, N-butylamino, 2-methoxyethoxy, 2-hydroxyethylamino, N-di (2-methoxyethyl) amino.

Another preferred embodiment of the present invention is:

wherein R is¹Each independently represents hydrogen or methyl;

R²each independently represents cyclopropyl, cyclohexyl, cyclopentyl, cycloheptylPhenyl, furanylpyridyl, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, phenoxy, cyclopentylamino, cyclopentylmethoxy or benzyloxy;

A¹and (b) represents NH;

A²each independently represents a bond or CH₂；

Q¹Each independently represents a phenyl group;

Q²is an aliphatic heterocycle selected from: tetrahydropyrrolyl, morpholinyl, N-methylpiperazinyl, piperazinyl;

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-phenylpyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-1)

4- ((6-cyclopentylpyrimidin-4-yl) amino) -N- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -1H-pyrazole-3-carboxamide (I-2)

4- ((6-cycloheptylpyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-3)

4- ((6-cyclopentylpyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-4)

4- ((6-Cyclopropylpyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-5)

4- ((6-Cyclopentylpyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-6)

4- ((6-cyclopentylpyrimidin-4-yl) amino) -N- (4- (morpholinomethyl) phenyl) -1H-pyrazole-3-carboxamide (I-7)

4- ((6- (furan-3-yl) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-8)

4- ((6- (furan-2-yl) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-9)

4- ((6-cyclopentylpyrimidin-4-yl) amino) -N- (4- (pyrrolidin-1-ylmethyl) phenyl) -1H-pyrazole-3-carboxamide (I-10)

4- ((4-cyclopentyl-6-methylpyrimidin-2-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-11)

4- ((4-cyclopentyl-6-methylpyrimidin-2-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-12)

4- ((4-cyclopentylpyrimidin-2-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-13)

N- (4-morpholino) -4- ((6-phenylpyrimidin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-14)

4- ((4-Cyclopentylpyrimidin-2-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-15)

4- ((6-cyclopentylpyrimidin-4-yl) amino) -N- (4- (4-methylpiperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-16)

4- ((2-methyl-6-phenoxypyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-17)

4- ((6- (cyclohexyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-18)

4- ((6- (cycloheptyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-19)

4- ((6- (2-bromophenoxy) pyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-20)

4- ((6- (cyclopentylmethoxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-21)

4- ((6- (cyclopentylamino) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-22)

4- ((6- (cyclopentyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-23)

4- ((4-phenoxypyrimidin-2-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-24)

4- ((6- (2-bromophenoxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-25)

4- ((4-methyl-6-phenoxypyrimidin-2-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-26)

4- ((6-phenoxypyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-27)

N- (4-morpholino) -4- ((6-phenoxypyrimidin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-28)

4- ((6- (cyclohexyloxy) pyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-29)

4- ((6- (Cyclopentyloxy) pyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-30)

4- ((6- (cyclopentylamino) pyrimidin-4-yl) amino) -N- (4-morpholinophenyl) -1H-pyrazole-3-carboxamide (I-31)

N- (4- ((4-methylpiperazin-1-yl) methyl) phenyl) -4- ((6-phenoxypyrimidin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-32)

N- (4- (morpholinomethyl) phenyl) -4- ((6-phenoxypyrimidin-4-yl) amino) -1H-pyrazole-3-carboxamide (I-33)

4- ((6- (benzyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-34) moieties of the invention

The preparation method of the compound 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 CDK2, CDK4, CDK6 and FLT3, so that the compound shown in the general formula I and the pharmaceutically acceptable salt thereof can be used for treating clinical symptoms related to the kinases. 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 their inhibitory activity against FLT3, CDK2, 4, 6 by Fluorescence Resonance Energy Transfer (FRET) method, and compounds with better activity were selected as compared with positive control. 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 appropriate concentration with a kinase diluent and used. The kinase reaction mixture contains FLT3, peptespbstrate, HEPES (pH 7.5), BRIJ-35, MgCl₂And 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. The reaction was continued at room temperature for 1h and stopped by the addition of Stop Reagent. Excitation wavelength 400nm and detection wavelength 445nFluorescence intensities of m (coumarin) and 520nm (fluoroescein). 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)^-6mol/L)

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

The MTT method is used for determining the inhibition effect on tumor cell strains such as a gastric cancer cell strain MGC803, a leukemia cell strain K562, a breast cancer cell strain MCF7, a leukemia cell strain MV4-11, a lung cancer cell strain A549, a colon cancer cell strain HCT116 and the like.

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. Then, the purple crystal in the cells is dissolved by dimethyl sulfoxide (DMSO) or triple liquid (10% SDS-5% isobutanol-0.01 mol/L HCL), and the OD value of the purple crystal is measured by an enzyme-linked immunosorbent assay detector at the wavelength of 570nm to indirectly reflect the living cell amount.

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 according to a certain cell amount, culturing for 24h, adding the screened sample (directly adding the suspension cells after connecting the plate), and culturing the cells at 37 ℃ with 5% CO₂After 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: leukemias, lymphomas (non-hodgkin lymphoma), hodgkin's disease (also known as hodgkin lymphoma) and myelomas such as, for example, Acute Lymphocytic Leukemia (ALL), acute myeloblastic or Acute Myeloid Leukemia (AML), Acute Promyelocytic Leukemia (APL), Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML), Chronic Neutrophilic Leukemia (CNL), Acute Undifferentiated Leukemia (AUL), operational developmental large cell lymphoma (ALCL), adult T-cell ALL, AML with trilineage (trilineage) myelodysplasia (AML/TMDS), Mixed Lineage Leukemia (MLL), myelodysplastic syndrome (MDSs), Myelodysplasia (MPD), Multiple Myeloma (MM) and myxosarcoma, lung cancer, melanoma, liver cancer, kidney cancer, leukemia, non-small cell lung cancer, leukemia, myelodysplasia, and myelosarcoma, Prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndromes, lymphomas, esophageal cancer, thyroid follicular cancer, gastrointestinal cancer, tumors of the central or peripheral nervous system (e.g., astrocytomas, neuroblastomas, gliomas, or schwannomas), mesotheliomas, type II or non-insulin dependent diabetes, autoimmune diseases.

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;¹HNMR was performed using JEOL FX90Q model Fourier transform nuclear magnetic resonance apparatus, BRUKER ACF-300 model 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)

In 500mL10g (46.3mmol) of p-nitrobenzyl bromide and 100mL of dichloromethane are added into a single-neck bottle, a mixed solution of 4.7g (47.0mmol) of N-methylpiperazine and 7.1g (70.3mmol) of triethylamine in 20mL of dichloromethane is slowly added dropwise under an ice water bath (0-5 ℃), and after the addition, the mixture is heated and refluxed for 1h, and the raw materials disappear 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 30 min. 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 distilled off under reduced pressure to obtain 8.5g of light yellow solid, the yield is 78.1 percent, and the product is directly put into the next reaction without further purification.¹H NMR(300MHz，DMSO)δ8.1(d，J＝8.6Hz，2H，ArH)，7.5(d，J＝8.6Hz，2H，ArH)，3.5(s，2H，-CH₂-)，2.3-2.5(br，8H，-CH₂-×4)，2.15(s，3H，-CH₃).

Example 2

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

A500 mL single-necked flask was charged with 8.5g (36.2mmol) 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 thereto to detect the disappearance of 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 (30mL 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.¹H NMR(300MHz，DMSO)δ8.1(d，J＝8.5Hz，2H，ArH)，7.5(d，J＝8.5Hz，2H，ArH)，4.0(s，2H，-NH₂)，3.5(s，2H，-CH₂-)，2.3-2.5(br，8H，-CH₂-×4)，2.1(s，3H，-CH₃)

Example 3

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

A250 mL round bottom flask was charged with 7.5g (36.6mmol) of crude I-a', 6.3g (40.1mmol) of 4-nitro-1H-pyrazole-3-carboxylic acid, 8.4g (44.0mmol) of EDC. HCl, 6.0g (44.4mmol) of HOBt and dry DMF100mL and stirred at room temperature for 24H. TLC detection of disappearance of starting Material (MeOH)1: 10 of chloroform). 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 11.1g of (I-e), wherein the yield is 88.2%. MS [ M + H ]]⁺345.3。¹H 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，-CH₂-)，2.3-2.4(br，8H，-CH₂-×4)，2.2(s，3H，-CH₃).

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-m 6.0g (17.4mmol), 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 the starting material (methanol: chloroform: 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30mL × 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。¹HNMR(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，-NH₂)，3.3(s，2H，-CH₂-)，2.3-2.5(m，8H，-CH₂-×4)，2.1(s，3H，-CH₃).

Example 5

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

10g (70.9mmol) of p-fluoronitrobenzene and 14.6g (106.4mmol) 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 (106.4mmol) in DMSO was slowly added dropwise under an ice-water bath (0-5 ℃ C.), and after the addition, the mixture was heated under reflux for 6 hours, and 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.¹H NMR(300MHz，DMSO)δ8.14(d，J＝8.7Hz，2H，ArH)，7.85(d，J＝8.7Hz，2H，ArH)，3.45(s，4H，-CH₂-×2)，3.39(s，4H，-CH₂-×2)，1.35(s，9H，-CH₃×3)。

Example 6

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

A500 mL single-necked flask was charged with 10g of crude I-e (32.6mmol), 1.0g of FeO (OH)/C catalyst, and 100mL of 95% ethanol, heated under reflux, and a mixture of 10mL of hydrazine hydrate and 20mL of 95% ethanol was slowly added dropwise thereto, followed by TLC detection of disappearance of the starting material (methanol: chloroform: 1: 15). The mixture is filtered while the mixture is hot, the filter cake is washed twice with hot ethanol (30mL 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-f), wherein the yield is 87.1%. The product is directly put into the next reaction without further purification.¹H NMR(300MHz，DMSO)δ6.94(d，J＝8.7Hz，2H，ArH)，6.22(s，2H，-NH₂)，6.15(d，2H，J＝8.7Hz，ArH)，3.45(t，4H，-CH₂-×2)，3.39(t，4H，-CH₂-×2)，1.35(s，9H，-CH₃×3)，

Example 7

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

A250 mL round bottom flask was charged with crude I-f 5g (18.1mmol), 4-nitro-1H-pyrazole-3-carboxylic acid 3.1g (19.9mmol), EDC. HCl 4.1g (21.7mmol), HOBt 2.9g (21.7mmol) and anhydrous DMF50mL 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-g), wherein the yield is 62.4%. MS [ M + H ]]⁺417.2.¹H 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，-CH₂-×2)，3.39(t，4H，-CH₂-×2)，1.45(s，9H，-CH₃×3)。

Example 8

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

A250 mL single-necked flask was charged with I-g 5.0g (12.0mmol), FeO (OH)/C catalyst 0.5g and 95% ethanol 60mL, heated to reflux, and hydrazine hydrate 5mL was slowly added dropwise to remove the starting material by TLC (methanol: chloroform: 1: 10). Filtering while hot, washing the filter cake twice with hot ethanol (30mL × 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)3.9g, yield 85.3%, and MS [ M + H ]]⁺386.2.¹H 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，-NH₂)，6.45(d，J＝8.7Hz，2H，ArH)，3.55(t，4H，-CH2-×2)，3.49(t，4H，-CH₂-×2)，1.45(s，9H，-CH₃×3)。

Example 9

Adding I-H150 mg (0.39mmol), 4-chloro-6-phenylpyrimidine 88.9mg (0.47mmol) and 50% aqueous acetic acid 5mL into a 50mL single-neck bottle, heating and refluxing, detecting by TLC that the raw materials disappear (methanol: chloroform: 1: 10), cooling the reaction solution to room temperature, adjusting the pH to 8-9 with saturated aqueous sodium bicarbonate solution, precipitating a solid, carrying out suction filtration, extracting the filtrate with ethyl acetate for 3 times (50 mL. times.3), combining the extracts, drying over anhydrous magnesium sulfate, carrying out suction filtration, distilling off the solvent under reduced pressure, combining with the filter cake, carrying out column chromatography (methanol: chloroform: 1: 15) on the crude product to obtain (I-1)107mg, obtaining the yield of 63%, [ M + H ], (M-1)]⁺441.2.¹H NMR(300MHz，DMSO)δ13.44(s，1H，pyrazole)，10.08(s，1H，-NH-)，9.51(s，1H，-NHCO-)，8.78(s，1H，ArH)，8.53(s，1H，ArH)，8.16(d，J＝3.7Hz，2H，ArH)，7.71(m，3H，ArH)，7.52(d，J＝3.5Hz，3H，ArH)，6.91(d，J＝8.7Hz，2H，ArH)，3.45(s，1H，-NH-)，3.01(s，4H，-CH₂-×2)，2.83(s，4H，-CH₂-×2).

Example 10

Adding I-d 150mg (0.48mmol), 4-chloro-6-cyclopentylpyrimidine 104mg (0.58mmol) and 50% aqueous acetic acid 5mL into a 50mL single-neck bottle, heating and refluxing, detecting by TLC that the raw materials disappear (methanol: chloroform: 1: 10), cooling the reaction solution to room temperature, adjusting the pH to 8-9 with saturated aqueous sodium bicarbonate solution, precipitating a solid, carrying out suction filtration, extracting the filtrate 3 times with ethyl acetate (50 mL. times.3), combining the extracts, drying over anhydrous magnesium sulfate, carrying out suction filtration, distilling off the solvent under reduced pressure, combining with the filter cake, carrying out column chromatography on the crude product (methanol: chloroform: 1: 15) to obtain (I-2)157mg, a light yellow solid, the yield being 71%, and MS [ M + H ], (M-chloroform: 1: 15)]⁺460.3.¹H NMR(300MHz，DMSO)δ13.40(s，1H，pyrazole)，10.19(s，1H，-NH-)，9.20(s，1H，-NHCO-)，8.60(s，1H，ArH)，8.45(s，1H，ArH)，7.79(d，J＝8.4Hz，2H，ArH)，7.25(d，J＝8.4Hz，2H，ArH)，6.96(s，1H，ArH)，3.45(s，2H，-CH₂-)，3.00(m，1H，-CH-)，2.48(dd，J＝13.1，11.4Hz，8H，-CH₂-×4)，2.28(s，3H，-CH₃)，1.95(dd，J＝16.6，8.4Hz，2H，-CH₂-)，1.69(m，6H，-CH₂-×3).

Example 11

In a similar manner to preparation of I-1, pale yellow solid (I-3) was obtained in 43% yield and MS [ M + H ]]⁺461.3.¹H NMR(300MHz，DMSO)δ13.46(s，1H，pyrazole)，10.09(s，1H，-NH-)，9.25(s，1H，-NHCO-)，8.59(s，1H，ArH)，8.43(s，1H，ArH)，7.73(d，J＝8.8Hz，2H，ArH)，6.96(m，3H，ArH)，3.35(s，4H，-CH₂-×2)，3.20(s，4H，-CH₂-×2)，3.15(s，1H，-NH-)，2.67(s，1H，-CH-)，1.68(m，14H，-CH₂-×7).

Example 12

In a similar manner to preparation of I-1, a pale yellow solid (I-4) was obtained in 44% yield, MS [ M + H%]⁺432.2.¹H NMR(300MHz，DMSO)δ13.42(s，1H，pyrazole)，10.09(s，1H，-NH-)，9.27(s，1H，-NHCO-)，9.07(s，1H，ArH)，8.61(s，1H，ArH)，8.44(s，1H，ArH)，7.73(d，J＝8.8Hz，2H，ArH)，6.97(m，3H，ArH)，3.33(s，4H，-CH₂-×2)，3.23(s，4H，-CH₂-×2)，3.02(m，-NH-)，2.9-3.0(m，1H，-CH-)，1.90(m，2H，-CH₂-)，1.73(s，6H，-CH₂-×3).

Example 13

In a similar manner to preparation of I-1, a pale yellow solid (I-5) was obtained in a yield of 38%, MS [ M + H ]]⁺405.2.¹H NMR(300MHz，DMSO)δ13.4(s，1H，pyrazole)，9.96(s，1H，-NH-)，9.21(s，1H，-NHCO-)，8.45(d，J＝26.5Hz，2H，ArH)，7.66(d，J＝8.9Hz，2H，ArH)，6.91(m，3H，ArH)，3.17(s，1H，-NH-)，3.02(m，4H，-CH₂-×2)，2.85(d，J＝4.8Hz，4H，-CH₂-×2)，1.92(m，1H，-CH-)，0.95(d，J＝7.4Hz，4H，-CH₂-×2).

Example 14

In a similar manner to preparation of I-1, pale yellow solid (I-6) was obtained in a yield of 54%, MS [ M + H ]]⁺434.2.¹H NMR(300MHz，DMSO)δ13.33(s，1H，pyrazole)，10.01(s，1H，-NH-)，9.26(s，1H，-NHCO-)，8.60(s，1H，ArH)，8.44(s，1H，ArH)，7.69(d，J＝8.9Hz，2H，ArH)，6.93(m，3H，ArH)，3.74(m，4H，-CH₂-×2)，3.07(m，4H，-CH₂-×2)，2.9-3.00(m，1H，-CH-)，1.93(s，2H，-CH₂-)，1.69(m，6H，-CH₂-×3).

Example 15

In a similar manner to that for the preparation of I-2, a pale yellow solid (I-7) was obtained in a yield of 56%, MS [ M + H ]]⁺448.2.¹H NMR(300MHz，DMSO)δ13.38(s，1H，pyrazole)，10.19(s，1H，-NH-)，9.21(s，1H，-NHCO-)，8.60(s，1H，ArH)，8.46(s，1H，ArH)，7.79(d，J＝8.4Hz，2H，ArH)，7.26(d，J＝8.4Hz，2H，ArH)，6.97(s，1H，ArH)，3.57(m，4H，-CH₂-×2)，3.42(s，2H，-CH₂-)，2.96(dd，J＝22.1，14.2Hz，1H，-CH-)，2.34(s，4H，-CH2-×2)，1.95(dd，J＝16.6，8.5Hz，2H，-CH₂-)，1.70(dt，J＝20.4，7.8Hz，6H，-CH₂-×3).

Example 16

In a similar manner to preparation of I-1, a pale yellow solid (I-8) was obtained in 52% yield, MS [ M + H]⁺431.2.¹H NMR(300MHz，DMSO)δ10.05(s，1H)，9.45(s，1H)，8.67(s，1H)，8.48(s，1H)，8.39(s，1H)，7.81(s，1H)，7.68(d，J＝9.0Hz，2H)，7.40(s，1H)，7.08(s，1H)，6.91(d，J＝9.1Hz，2H)，3.15(m，1H，-NH-)，3.02(d，J＝5.1Hz，4H)，2.86(s，4H).

Example 17

In a similar manner to preparation of I-1, a pale yellow solid (I-9) was obtained in a yield of 46%, MS [ M + H ]]⁺431.2.¹H NMR(300MHz，DMSO)δ13.4(s，1H，pyrazole)，10.09(s，1H)，9.38(s，1H)，8.66(s，1H)，8.48(s，1H)，8.13(s，1H)，7.94(s，1H)，7.69(d，J＝9.0Hz，2H)，7.37(s，1H)，7.05(s，1H)，6.88(d，J＝9.1Hz，2H)，3.07(m，1H，-NH-)，3.00(d，J＝5.1Hz，4H)，2.76(s，4H).

Example 18

In a similar manner to that for the preparation of I-2, a pale yellow solid (I-10) was obtained in a yield of 61%, MS [ M + H ]]⁺432.2.¹H NMR(300MHz，DMSO)δ13.45(s，1H，pyrazole)，10.14(s，1H，-NH-)，9.07(s，1H，-NHCO-)，8.67(s，1H，ArH)，8.55(s，1H，ArH)，7.69(d，J＝8.4Hz，2H，ArH)，7.15(d，J＝8.4Hz，2H，ArH)，6.91(s，1H，ArH)，3.42(s，2H，-CH₂-)，3.15(m，1H，-CH-)，2.85(s，4H，-CH₂-×2)，2.15(s，4H，-CH₂-×2)，1.82-1.86(m，2H，-CH₂-)，1.69(m，6H，-CH₂-×3).

Example 19

In a similar manner to that for the preparation of I-2, a pale yellow solid (I-11) was obtained in a yield of 67%, MS [ M + H ]]⁺447.3.¹H NMR(300MHz，DMSO)δ13.44(s，1H，pyrazole)，10.14(s，1H，-NH-)，9.31(s，1H，-NHCO-)，9.12(s，1H，ArH)，8.14(s，1H，ArH)，7.69(d，J＝8.8Hz，2H，ArH)，6.95-7.04(m，3H，ArH)，3.31(s，4H，-CH₂-×2)，3.21(s，4H，-CH₂-×2)，3.02(m，-NH-)，2.9-3.0(m，1H，-CH-)，2.65(s，3H，-CH₃)，1.94(m，2H，-CH₂-)，1.81(s，6H，-CH₂-×3).

Example 20

In a similar manner to that for the preparation of I-1, a pale yellow solid (I-12) was obtained in a yield of 45%, MS [ M + H%]⁺419.5.¹H NMR(300MHz，DMSO)δ13.19(s，1H，pyrazole)，10.21(s，1H，-NH-)，9.45(s，1H，-NHCO-)，8.37(d，J＝8.9Hz，2H，ArH)，7.62(d，J＝8.9Hz，2H，ArH)，6.92(m，3H，ArH)，3.17(s，1H，-NH-)，3.12(m，4H，-CH₂-×2)，2.94(m，4H，-CH₂-×2)，2.82(m，1H，-CH-)，2.73(s，3H，-CH₃)，0.88(d，J＝7.4Hz，4H，-CH₂-×2).

Example 21

In a similar manner to that for the preparation of I-1, a pale yellow color is obtainedSolid (I-13) in 42% yield MS [ M + H%]⁺433.2.¹H NMR(300MHz，DMSO)δ13.33(s，1H，pyrazole)，10.12(s，1H，-NH-)，9.35(s，1H，-NHCO-)，8.66(s，1H，ArH)，8.24(s，1H，ArH)，7.68(d，J＝8.8Hz，2H，ArH)，6.95(m，3H，ArH)，3.23(s，4H，-CH₂-×2)，3.09(s，4H，-CH₂-×2)，3.04(m，-NH-)，2.7-2.9(m，1H，-CH-)，1.88(m，2H，-CH₂-)，1.67(s，6H，-CH₂-×3).

Example 22

In a similar manner to preparation of I-1, a pale yellow solid (I-14) was obtained in a yield of 47%, MS [ M + H ]]⁺442.2.¹H NMR(300MHz，DMSO)δ13.42(s，1H，pyrazole)，10.05(s，1H，-NH-)，9.44(s，1H，-NHCO-)，8.76(s，1H，ArH)，8.47(s，1H，ArH)，8.15(d，J＝3.7Hz，2H，ArH)，7.65(m，3H，ArH)，7.49(d，J＝3.5Hz，3H，ArH)，6.92(d，J＝8.7Hz，2H，ArH)，3.65(s，4H，-CH₂-×2)，2.98(s，4H，-CH₂-×2).

Example 23

In a similar manner to preparation of I-1, a pale yellow solid (I-15) was obtained in 55% yield and MS [ M + H ]]⁺434.2.¹H NMR(300MHz，DMSO)δ13.45(s，1H，pyrazole)，10.03(s，1H，-NH-)，9.22(s，1H，-NHCO-)，8.44(s，1H，ArH)，8.36(s，1H，ArH)，7.69(d，J＝8.8Hz，2H，ArH)，6.92(m，3H，ArH)，3.69(s，4H，-CH₂-×2)，3.10(s，4H，-CH₂-×2)，2.81-2.96(m，1H，-CH-)，1.92(m，2H，-CH₂-)，1.72-1.76(m，6H，-CH₂-×3).

Example 24

In a similar manner to that for preparation of I-1, a pale yellow solid (I-18) was obtained in a yield of 29%，¹H NMR(300MHz，DMSO)δ13.42(s，1H，pyrazole)，10.09(s，1H，-NH-)，9.27(s，1H，-NHCO-)，8.61(s，1H，ArH)，8.40(s，1H，ArH)，7.71(d，J＝8.4Hz，2H，ArH)，7.28(d，J＝8.4Hz，2H，ArH)，7.05(s，1H，ArH)，3.87(s，4H，-CH₂-×2)，3.01(s，4H，-CH₂-×2)，2.56(s，3H，-CH₃)，1.95(dd，J＝16.6，8.4Hz，2H，-CH₂-)，1.69(m，6H，-CH₂-×3).

Example 25

In a similar manner to preparation of I-1, a pale yellow solid (I-17) was obtained in a yield of 25%, MS [ M + H ]]⁺471.2.¹H NMR(300MHz，DMSO)δ13.47(s，1H，pyrazole)，10.15(s，1H，-NH-)，9.33(s，1H，-NHCO-)，8.51(s，1H，ArH)，7.79(d，J＝8.9Hz，2H，ArH)，7.52(t，J＝7.8Hz，2H，ArH)，7.33(t，J＝7.4Hz，1H，ArH)，7.23(d，J＝7.6Hz，2H，ArH)，7.04(d，J＝9.0Hz，2H，ArH)，6.35(s，1H，ArH)，3.39(s，4H，-CH₂-×2)，3.12(s，4H，-CH₂-×2)，3.14(m，1H，-NH-)，2.45(s，3H，-CH₃).

Example 26

In a similar manner to preparation of I-1, a pale yellow solid (I-18) was obtained in 19% yield, MS [ M + H]+463.2.¹HNMR(300MHz，DMSO)δ13.32(br，1H，pyrazole)，9.96(s，1H，-NH-)，9.14(s，1H，ArH)，8.37(s，2H，ArH)，7.65(d，J＝8.9Hz，2H，ArH)，6.90(d，J＝8.9Hz，2H，ArH)，6.31(s，1H，ArH)，4.95(m，1H，-CH-)，3.04(m，4H，-CH₂-×2)，2.88(s，4H，-CH₂-×2)，1.91(s，1H，-NH-)，1.72(m，2H，-CH₂-)，1.34-1.41(m，4H，-CH₂-×2)，1.20(s，4H，-CH₂-×2).

Example 27

In a similar manner to preparation of I-1, a pale yellow solid (I-19) was obtained in a yield of 17%, MS [ M + H ]]+477.3.¹HNMR(300MHz，DMSO)δ13.31(s，1H，pyrazole)，9.97(s，1H，-NH-)，9.13(s，1H，-NHCO-)，8.37(s，2H，ArH)，7.66(d，J＝9.1Hz，2H，ArH)，6.91(d，J＝9.0Hz，2H，ArH)，6.29(s，1H，ArH)，5.14(m，1H，-CH-)，3.08(s，4H，-CH₂-×2)，2.94(s，4H，-CH₂-×2)，1.98(m，1H，-NH-)，1.66(br，4H，CH₂-×2)，1.55(br，4H，-CH₂-×2)，1.44(br，2H，-CH₂-)，1.23(m，2H，-CH₂-).

Example 28

In a similar manner to that for the preparation of I-1, a pale yellow solid (I-20) was obtained in a yield of 30%, MS [ M + H ]]⁺536.1.¹H NMR(300MHz，DMSO)δ13.36(s，1H，pyrazole)，10.03(s，1H，-NH-)，9.40(s，1H，-NHCO-)，8.43(s，1H，ArH)，8.35(s，1H，ArH)，7.72(dd，J＝13.9，7.7Hz，3H，ArH)，7.46(d，J＝7.3Hz，1H，ArH)，7.33(d，J＝6.8Hz，1H，ArH)，7.29-7.21(m，1H，ArH)，6.92(d，J＝8.9Hz，2H，ArH)，6.64(s，1H，ArH)，3.79-3.69(m，4H，-CH₂-×2)，3.07(s，4H，-CH₂-×2).

Example 29

In a similar manner to preparation of I-1, a pale yellow solid (I-21) was obtained in 19% yield, MS [ M + H]⁺463.2.¹H NMR(300MHz，DMSO)δ13.36(s，1H，pyrazole)，10.06(s，1H，-NH-)，9.16(s，1H，-NHCO-)，8.87-8.78(m，1H，ArH)，8.37(s，1H，ArH)，7.72(d，J＝6.5Hz，2H，ArH)，6.98(d，J＝7.4Hz，2H，ArH)，6.36(m，1H，ArH)，3.23(br，4H，-CH₂-×2)，3.17(s，1H，-CH-)，3.08(d，4H，-CH₂-×2)，2.28(m，2H，-OCH₂-)，1.92(s，1H，-NH-)，1.57-1.73(m，8H，-CH₂-×4).

Example 30

In a similar manner to preparation of I-1, a pale yellow solid (I-22) was obtained in 21% yield, MS [ M + H%]⁺448.2.¹H NMR(300MHz，DMSO)δ13.27(s，1H，pyrazole)，9.98(s，1H，-NH-)，9.60(s，1H，-NHCO-)，8.78(s，1H，ArH)，8.22(s，1H，ArH)，8.12(s，1H，ArH)，7.66(d，J＝8.9Hz，2H，ArH)，6.92(d，J＝8.9Hz，2H，ArH)，6.83(d，J＝7.3Hz，1H，ArH)，5.85(m，1H，-NCH-)，3.09(s，4H，-CH₂-×2)，2.94(s，4H，-CH₂-×2)，2.62(m，1H，-NH-)，2.11(s，1H，-NH-)，1.91(br，2H，-CH₂-)，1.67(br，2H，-CH₂-)，1.44-1.54(m，4H，-CH₂-×2).

Example 31

In a similar manner to preparation of I-1, a pale yellow solid (I-23) was obtained in a yield of 15%, MS [ M + H ]]⁺449.2.¹H NMR(300MHz，DMSO)δ13.38(s，1H，pyrazole)，10.07(s，1H，-NH-)，9.14(s，1H，-NHCO-)，8.85(s，1H，ArH)，8.38(s，1H，ArH)，7.70(s，2H，ArH)，6.96(s，2H，ArH)，6.32(s，1H，ArH)，5.29(s，1H，-CH-)，3.17(s，4H，-CH₂-×2)，2.46(s，4H，-CH₂-×2)，1.90(s，1H，-NH-)，1.67(br，6H，-CH₂-×3)，1.18(s，2H，-CH₂-).

Example 32

In a similar manner to that for the preparation of I-1, a pale yellow solid (I-24) was obtained in a yield of 24%, MS [ M + H ]]⁺471.2.¹H NMR(300MHz，DMSO)δ13.25-12.90(s，1H，pyrazole)，9.93(s，1H，-NH-)，9.10(s，1H，-NHCO-)，8.23(d，J＝11.0Hz，1H，ArH)，7.62(d，J＝9.1Hz，2H，ArH)，7.54(t，J＝7.7Hz，2H，ArH)，7.39(d，J＝7.2Hz，1H，ArH)，7.27(d，J＝7.6Hz，2H，ArH)，6.88(d，J＝9.0Hz，3H，ArH)，3.04(d，J＝5.2Hz，4H，-CH₂-×2)，2.91(d，J＝5.2Hz，4H，-CH₂-×2)，2.11(s，1H，-NH-).

Example 33

In a similar manner to preparation of I-1, a pale yellow solid (I-25) was obtained in 19% yield, MS [ M + H%]⁺535.1.¹H NMR(300MHz，DMSO)δ13.36(s，1H，pyrazole)，10.03(s，1H，-NH-)，9.40(s，1H，-NHCO-)，8.43(s，1H，ArH)，8.35(s，1H，ArH)，7.72(dd，J＝13.9，7.7Hz，3H，ArH)，7.46(d，J＝7.3Hz，1H，ArH)，7.23(d，J＝6.8Hz，1H，ArH)，7.29-7.21(m，1H，ArH)，6.92(d，J＝8.9Hz，2H，ArH)，6.64(s，1H，ArH)，3.08(s，4H，-CH₂-×2)，2.94(s，4H，-CH₂-×2)，2.23(s，1H，-NH-).

Example 34

In a similar manner to that for the preparation of I-1, a pale yellow solid (I-26) was obtained in a yield of 17%, MS [ M + H%]⁺471.2.¹H NMR(300MHz，DMSO)δ13.25-12.90(s，1H，pyrazole)，9.93(s，1H，-NH-)，9.10(s，1H，-NHCO-)，8.23(d，J＝11.0Hz，1H，ArH)，7.62(d，J＝9.1Hz，2H，ArH)，7.54(t，J＝7.7Hz，2H，ArH)，7.39(d，J＝7.2Hz，1H，ArH)，7.27(d，J＝7.6Hz，2H，ArH)，6.88(d，J＝9.0Hz，2H，ArH)，6.38(s，1H，ArH)，3.04(d，J＝5.2Hz，4H，-CH₂-×2)，2.91(d，J＝5.2Hz，4H，-CH₂-×2)，2.34(s，3H，-CH₃)，2.20(s，1H，-NH-).

Example 35

In a similar manner to preparation of I-1, a pale yellow solid (I-27) was obtained in a yield of 19%, MS [ M + H ]]⁺457.2.¹H NMR(300MHz，DMSO)δ13.32(s，1H，pyrazole)，10.33(s，1H，-NH-)，9.12(s，1H，-NHCO-)，8.45(s，1H，ArH)，8.31(d，J＝10.0Hz，1H，ArH)，7.71(d，J＝9.3Hz，2H，ArH)，7.64(t，J＝7.7Hz，2H，ArH)，7.41(d，J＝7.2Hz，1H，ArH)，7.37(d，J＝7.6Hz，2H，ArH)，6.81(d，J＝9.0Hz，2H，ArH)，6.37(s，1H，ArH)，3.14(br，4H，-CH₂-×2)，2.93(br，4H，-CH₂-×2)，2.17(s，1H，-NH-).

Example 36

In a similar manner to preparation of I-1, a pale yellow solid (I-28) was obtained in 19% yield, MS [ M + H]⁺458.1.¹H NMR(300MHz，DMSO)δ13.34(s，1H，pyrazole)，10.02(s，1H，-NH-)，9.35(s，1H，-NHCO-)，8.44(s，1H，ArH)，8.37(s，1H，ArH)，8.21(s，1H，ArH)，7.69(m，3H，ArH)，7.41(d，J＝7.3Hz，1H，ArH)，7.32(s，1H，ArH)，7.26-7.29(m，1H，ArH)，6.94(d，J＝8.9Hz，2H，ArH)，6.65(s，1H，ArH)，3.77-3.68(m，4H，-CH₂-×2)，3.08(br，4H，-CH₂-×2).

Example 37

In a similar manner to preparation of I-1, a pale yellow solid (I-29) was obtained in 19% yield, MS [ M + H]⁺464.2.¹H NMR(300MHz，DMSO)δ13.31(br，1H，pyrazole)，10.03(s，1H，-NH-)，9.24(s，1H，-NHCO-)，8.33(s，2H，ArH)，7.67(d，J＝8.9Hz，2H，ArH)，6.91(d，J＝8.9Hz，2H，ArH)，6.32(s，1H，ArH)，4.95(m，1H，-CH-)，3.14(m，4H，-CH₂-×2)，2.87(s，4H，-CH₂-×2)，1.72(m，2H，-CH₂-)，1.34-1.41(m，4H，-CH₂-×2)，1.20(s，4H，-CH₂-×2).

Example 38

In a similar manner to preparation of I-1, a pale yellow solid (I-30) was obtained in 19% yield, MS [ M + H%]⁺450.2.¹H NMR(300MHz，DMSO)δ13.35(s，1H，pyrazole)，10.04(s，1H，-NH-)，9.11(s，1H，-NHCO-)，8.34(s，1H，ArH)，8.27(s，1H，ArH)，7.73(s，2H，ArH)，6.98(s，2H，ArH)，6.34(s，1H，ArH)，4.06(s，1H，-CH-)，3.78(s，4H，-CH₂-×2)，3.12(s，4H，-CH₂-×2)，1.91(s，2H，-CH₂-)，1.68(br，6H，-CH₂-×3).

Example 39

In a similar manner to preparation of I-1, a pale yellow solid (I-31) was obtained in 24% yield, MS [ M + H%]⁺449.2.¹H NMR(300MHz，DMSO)δ13.31(s，1H，pyrazole)，9.96(s，1H，-NH-)，9.62(s，1H，-NHCO-)，8.20(s，1H，ArH)，8.11(s，1H，ArH)，7.69(d，J＝8.9Hz，2H，ArH)，6.91(d，J＝8.9Hz，2H，ArH)，6.82(d，J＝7.3Hz，1H，ArH)，4.56(s，1H，-NH-)，4.15(s，1H，-CH-)，3.78(br，4H，-CH₂-×2)，2.94(s，4H，-CH₂-×2)，1.92(br，2H，-CH₂-)，1.66(br，2H，-CH₂-)，1.41-1.54(m，4H，-CH₂-×2).

Example 40

In a similar manner to preparation of I-1, a pale yellow solid (I-32) was obtained in 19% yield, MS [ M + H%]⁺485.2.¹H NMR(300MHz，DMSO)δ13.36(s，1H，pyrazole)，10.31(s，1H，-NH-)，9.11(s，1H，-NHCO-)，8.42(s，1H，ArH)，8.33(d，J＝10.0Hz，1H，ArH)，7.69(d，J＝9.3Hz，2H，ArH)，7.65(t，J＝7.7Hz，2H，ArH)，7.43(d，J＝7.2Hz，1H，ArH)，7.36(d，J＝7.6Hz，2H，ArH)，6.82(d，J＝9.0Hz，2H，ArH)，6.36(s，1H，ArH)，3.44(s，2H，-CH₂-)，2.47(br，8H，-CH₂-×4)，2.17(s，2H，-CH₃).

EXAMPLE 41

In a similar manner to preparation of I-1, a pale yellow solid (I-33) was obtained in 21% yield, MS [ M + H%]⁺472.2.¹H NMR(300MHz，DMSO)δ13.32(s，1H，pyrazole)，10.23(s，1H，-NH-)，9.17(s，1H，-NHCO-)，8.44(s，1H，ArH)，8.31(d，J＝10.0Hz，1H，ArH)，7.70(d，J＝9.3Hz，2H，ArH)，7.64(t，J＝7.7Hz，2H，ArH)，7.41(d，J＝7.2Hz，1H，ArH)，7.37(d，J＝7.6Hz，2H，ArH)，6.81(d，J＝9.0Hz，2H，ArH)，6.34(s，1H，ArH)，3.44(s，2H，-CH₂-)，3.14(br，4H，-CH₂-×2)，2.46(br，4H，-CH₂-×2)，.

Example 42

4- ((6- (benzyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-34)

In a similar manner to preparation of I-1, a pale yellow solid (I-34) was obtained in 22% yield, MS [ M + H%]⁺471.2.¹H NMR(300MHz，DMSO)δ13.31(s，1H，pyrazole)，10.03(s，1H，-NH-)，9.36(s，1H，-NHCO-)，8.43(s，1H，ArH)，8.35(s，1H，ArH)，8.20(s，1H，ArH)，7.64(m，3H，ArH)，7.53(d，J＝7.3Hz，1H，ArH)，7.39(s，1H，ArH)，7.23(m，1H，ArH)，6.95(s，2H，ArH)，6.63(s，1H，ArH)，4.51(s，2H，-CH₂-)，3.66(m，4H，-CH₂-×2)，3.01(br，4H，-CH₂-×2)，2.14(s，1H，-NH-).

Claims

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

wherein R is¹Each independently represents hydrogen, methyl;

R²each independently represents cyclopropyl, cyclohexyl, cyclopentyl, cycloheptyl, phenyl, furyl, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, phenoxy, cyclopentylamino, cyclopentylmethoxy or benzyloxy;

A¹represents NH;

A²each independently represents a bond or CH₂；

Q¹Represents a phenyl group;

Q²is an aliphatic heterocycle selected from: tetrahydropyrrolyl, morpholinyl, N-methylpiperazinyl, piperazinyl.

2. The compound of claim 1, characterized in that it is the following compound:

4- ((6- (benzyloxy) pyrimidin-4-yl) amino) -N- (4- (piperazin-1-yl) phenyl) -1H-pyrazole-3-carboxamide (I-34).

3. A pharmaceutically acceptable salt of a compound according to any one of claims 1-2, wherein the pharmaceutically acceptable salt comprises an acid addition salt of a compound of formula (I): 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 1-2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a condition associated with FLT 3.

6. Use according to claim 5, wherein the disease associated with FLT3 is the following malignant proliferative disease: leukemia, myelodysplastic syndrome, or lymphoma.