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HK1200162B - Substituted indazole derivatives active as kinase inhibitors - Google Patents

Substituted indazole derivatives active as kinase inhibitors Download PDF

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Publication number
HK1200162B
HK1200162B HK15100468.6A HK15100468A HK1200162B HK 1200162 B HK1200162 B HK 1200162B HK 15100468 A HK15100468 A HK 15100468A HK 1200162 B HK1200162 B HK 1200162B
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HK
Hong Kong
Prior art keywords
indazol
methyl
benzamide
compound
formula
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HK15100468.6A
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Chinese (zh)
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HK1200162A1 (en
Inventor
A‧洛穆巴蒂波吉亚
M‧梅尼奇恩彻里
P‧奥西尼
A‧潘泽利
E‧派罗纳
E‧万诺提
M‧纳希
C‧玛奇昂尼
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内尔维阿诺医学科学有限公司
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Publication of HK1200162A1 publication Critical patent/HK1200162A1/en
Publication of HK1200162B publication Critical patent/HK1200162B/en

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Description

Substituted indazole derivatives active as kinase inhibitors
The application is a divisional application of Chinese patent application No. 200880025455.1 with the same name, the original international application number is PCT/EP2008/058861, and the international application date is 2008/7 and 8.
Technical Field
The present invention relates to certain substituted indazole compounds that modulate the activity of protein kinases. Accordingly, the compounds of the present invention are useful for the treatment of diseases caused by dysregulated protein kinase activity. The invention also provides methods of preparing these compounds, pharmaceutical compositions comprising these compounds, and methods of treating diseases utilizing pharmaceutical compositions comprising these compounds.
Background
Malfunction of Protein Kinases (PKs) is a hallmark of many diseases. Oncogenes and proto-oncogenes involved in human cancer encode a large proportion of PK. Enhanced PK activity is also associated with a number of non-malignant diseases, such as benign prostate hyperplasia, familial adenomatosis, polyposis, neurofibromatosis, psoriasis, vascular smooth cell proliferation associated with atherosclerosis, pulmonary fibrosis, arthritis, glomerulonephritis and post-surgical stenosis and restenosis.
PK is also associated with inflammatory conditions and the proliferation of viruses and parasites. PK may also play an important role in the pathogenesis and development of neurodegenerative diseases.
For general references on PK failure or malfunction, see, e.g., Current Opinion in chemical biology1999, 3, 459-.
One group of PKs is a group of membrane receptors with intrinsic protein tyrosine kinase activity (RPTK). Upon binding of growth factors, RPTK becomes activated and phosphorylates itself and a range of substrates in the cytoplasm. By this mechanism, they can transduce intracellular signaling for proliferation, differentiation, or other biological changes. Structural abnormalities, overexpression and activation of RTPKs are commonly observed in human tumors, suggesting that constitutive firing of signal transduction leading to cell proliferation may lead to malignant tumor transformation. Anaplastic lymphoma enzyme (ALK) is a tyrosine kinase receptor belonging to the insulin receptor subfamily of RTKs: the ALK gene is located on chromosome 2 and is expressed primarily on neuronal cells, particularly during development. The ALK gene is involved in a chromosomal translocation in Anaplastic Large Cell Lymphoma (ALCL) that is balanced with the Nuclear Phosphoprotein (NPM) gene on chromosome 5. In ALK + ALCL, the NPM ubiquitous promoter drives ectopic expression of fusion proteins due to translocation, where the NPM partially dimerizes and the ALK kinase domain autophosphorylates and becomes constitutively active.
Many data from the literature demonstrate that NPM-ALK fusion proteins have strong oncogenic potential and that ectopic expression thereof is responsible for cell transformation. Furthermore, constitutive expression of human NPM-ALK in mouse T cell lymphocytes is sufficient for the development of lymphoma formation in transgenic animals after a short latency period.
ALCL is a defined class of diseases characterized by the surface expression of the cell differentiation antigen CD30(Ki-1) and is responsible for 2% of adults and 13% of pediatric non-hodgkin's lymphomas, primarily affecting young male patients. ALK + ALCL accounts for 70% of all ALCLs and is an erosive disease with systemic signals and is a common extranodal involvement (bone marrow, skin, bone, soft tissue).
Approximately 15-20% of ALK-expressing ALCLs were found to carry a different chromosomal translocation, involving the cytoplasmic portion of ALK, with a different N-terminal portion, all of which resulted in constitutive activation of the ALK kinase domain.
In addition, cell lines established from solid tumors of ectodermal origin (e.g., melanoma, breast tumors, and neuroblastoma, glioblastoma, ewing's sarcoma, retinoblastoma) were found to express ALK receptors.
In summary, interfering with ALK signaling may represent a specific and effective approach for blocking tumor cell proliferation in ALCL and possibly other indications.
Insulin-like growth factor type 1 receptors (IGF-1R, IGF1R) are also members of the insulin receptor subfamily of RTKs.
There are several lines of evidence suggesting that IGF-1R signaling may contribute to tumorigenesis and interfere with IGF-1R function represents an effective therapeutic option for cancer. For a detailed description of reviews of IGF and IGF-1R signaling, physiological functions, and evidence supporting the association of this system in human cancer and other pathologies outlined above, the reader is referred to many reviews and references contained therein on this subject, e.g., Baserga R, et al, Biochim biophysicaca vol.1332, pages F105-F126, 1997; khandwala h.m. et al, Endocr Rev vol.21, pages 215-44, 2000; le Roith D, et al, Endocr Rev vol.22, pp 53-74, 2001; valentinis B.et al, Mol Patholvol.54, pp.133-7, 2001; wang Y, et al, Curr Cancer Drug Targets vol.2, pp.191-207, 2002; laron, Z.J Clin Endocrinol Metab vol.89, pp.1031-1044, 2004; hofmann F et al, drug Discov Today vol.10, pages 1041-7, 2005.
Disclosure of Invention
3-amino and 3-acylaminoindazole derivatives for the treatment of neurodegenerative diseases, cerebrovascular accidents, obesity, cardiovascular diseases and cancer are disclosed in WO2006003276, WO2004022544 and WO2003078403 in the name of Aventis Pharma SA.
Indazolylamide derivatives for the treatment of diabetes, neurodegenerative diseases, such as alzheimer's disease and parkinson's disease are disclosed in WO2003051847 in the name of SmithKline Beecham p.l.c.
In WO2008003396 in the name of Merck GMBH indazole derivatives for the treatment of tumor diseases, viral diseases, immunosuppression in transplantation, cystic fibrosis and angiogenesis-related diseases are disclosed.
Despite these developments, there remains a need for more effective agents for treating such diseases.
We have now found that a range of indazoles are potent inhibitors of protein kinases and are therefore useful in anticancer therapy.
Accordingly, a first aspect of the present invention provides a substituted indazole compound represented by formula (I),
wherein:
x is-CH2-, -CH (OH) -, -CH (OR ') -, OR-C (R ' R ') -, in which:
r' is optionally further substituted straight or branched chain C1-C6Alkyl, R' is hydrogen or optionally further substituted straight or branched chain C1-C6An alkyl group;
ar is aryl or heteroaryl optionally substituted with one or more substituents selected from: halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein:
r4 is hydrogen, C2-C6Alkenyl radical, C2-C6Alkynyl, NR5R6, OR7, SR7, R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substitutedSubstituted straight or branched chain C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl;
r5 and R6 are independently hydrogen, C2-C6Alkenyl radical, C2-C6Alkynyl, R8R9N-C2-C6Alkyl radical, R8O-C2-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, or R5 and R6 together with the bound nitrogen atom form an optionally substituted heterocyclyl;
r7 is hydrogen, C2-C6Alkenyl radical, C2-C6Alkynyl, COR4, SOR10, SO2R10，R8R9N-C2-C6Alkyl radical, R8O-C2-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein R4 is as defined above;
r8 and R9 are independently hydrogen, C2-C6Alkenyl radical, C2-C6Alkynyl, COR4, optionally further substituted straight or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, or R8 and R9 together with the bound nitrogen atom form an optionally substituted heterocyclyl, wherein R4 is as defined above;
r10 is hydrogen, C2-C6Alkenyl radical, C2-C6Alkynyl, NR5R6, OR7, R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein R5, R6, R7, R8 and R9 are as defined above;
r is optionally substituted straight or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl;
r1, R2 and R3 are independently hydrogen, halogen, nitro, optionally substituted straight or branched C1-C6Alkyl, NR5R6 OR7, wherein R5, R6 and R7 are as defined above;
or an isomer, tautomer, prodrug or pharmaceutically acceptable salt thereof.
The present invention also provides methods for the synthesis of substituted indazole derivatives of formula (I) prepared by a method consisting of standard synthetic transformations.
The invention also provides methods of treating diseases caused by and/or associated with dysregulated protein kinase activity, in particular the PLK family, the different isoforms of the protein kinases C, Met, PAK-4, PAK-5, ZC-1, STLK-2, DDR-2, Aurora1, Aurora2, Bub-1, Chk1, Chk2, HER2, raf1, MEK1, MAPK, EGF-R, PDGF-R, FGF-R, FLT3, JAK2, IGF-R, ALK, PI3K, wel kinases, Src, Abl, Akt, MAPK, ILK, MK-2, IKK-2, Cdc7, Nek, Cdk/cyclin kinase family, more particularly Aurora2, IGF-1R and ALK activity, and further in particular ALK activity, comprising administering to a mammal in need thereof an effective amount of a substituted indazole compound represented by formula (I) as defined above.
A preferred method of the invention is the treatment of a disease caused by and/or associated with dysregulated protein kinase activity, said disease being selected from the group consisting of cancer and cell proliferative disorders.
Another preferred method of the invention is the treatment of specific types of cancer, including hematopoietic tumors of myeloid or lymphoid lineage, tumors of mesenchymal origin, tumors of the central and peripheral nervous system, melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, angular warts of the cornea, thyroid follicular cancer and kaposi's sarcoma.
Another preferred method of the invention is the treatment of a particular type of cancer, such as, but not limited to, breast cancer, lung cancer, colorectal cancer, prostate cancer, ovarian cancer, endometrial cancer, gastric cancer, clear cell renal cell carcinoma, uveal melanoma, multiple myeloma, rhabdomyosarcoma, ewing's sarcoma, kaposi's sarcoma, and medulloblastoma.
Another preferred method of the invention is the treatment of ALK + Anaplastic Large Cell Lymphoma (ALCL) or other possible indications where ALK activity may play a role, such as neuroblastoma, rhabdomyosarcoma, glioblastoma, inflammatory myofibroma and certain types of melanoma, breast cancer, ewing's sarcoma, retinoblastoma and non-small cell lung cancer (NSCLC).
Another preferred method of the invention is the treatment of cell proliferative diseases such as, but not limited to, benign prostate hyperplasia, familial adenomatosis, polyposis, neurofibromatosis, psoriasis, atherosclerosis and diseases associated with vascular smooth cell proliferation and neointimal formation, such as restenosis after angioplasty or surgery, pulmonary fibrosis, arthritis, glomerulonephritis, retinopathies, including diabetic and neonatal retinopathy and age-related macular degeneration, graft vascular diseases (such as may occur following vascular or organ transplantation), acromegaly and conditions secondary to acromegaly, and other hypertrophic diseases involving IGF/IGF-1R signaling, such as fibrotic lung diseases, pathological conditions involving chronic or acute oxidative stress or hyperoxia-induced tissue damage, and metabolic disorders in which elevated IGF levels or IGF-1R activity are involved, such as obesity.
In addition, the methods of the invention also provide for inhibition of tumor angiogenesis and metastasis.
In another preferred embodiment, the methods of the invention further comprise subjecting the mammal in need thereof to a radiation or chemotherapy regimen in combination with at least one cytostatic or cytotoxic agent.
The present invention also provides a method of inhibiting an active ALK protein comprising contacting the protein with an effective amount of a compound of formula (I).
The invention also provides a pharmaceutical composition comprising one or more compounds of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, carrier or diluent.
The invention further provides pharmaceutical compositions comprising compounds of formula (I) in combination with one or more chemotherapeutic agents or radiation therapy. Such drugs may include, but are not limited to, anti-hormonal agents (e.g., antiestrogens, antiandrogens, and aromatase inhibitors), topoisomerase I inhibitors, topoisomerase II inhibitors, microtubule-targeting agents, platinum-based agents, alkylating agents, DNA damaging or intercalating agents, antitumor antimetabolites, other kinase inhibitors, other anti-angiogenic factors, inhibitors of kinesin, therapeutic monoclonal antibodies, inhibitors of mTOR, histone deacetylase inhibitors, farnesyl transferase inhibitors, and inhibitors of hypoxia response.
In addition, the present invention provides a product or kit comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above or a pharmaceutical composition thereof and one or more chemotherapeutic agents as a combined preparation for simultaneous, separate or sequential use in anticancer therapy.
In another aspect, the present invention provides a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, for use as a medicament.
The invention also provides the use of a compound of formula (I) as defined above, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament having anti-tumour activity.
Finally, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined above, in a method of treating cancer.
Detailed Description
The compounds of formula (I) may have one or more asymmetric centers and may thus exist as individual optical isomers or as racemic mixtures. Thus, all possible isomers of the compounds of formula (I), as well as mixtures thereof, are within the scope of the present invention.
Derivatives of the compounds of formula (I) that are metabolically derived in mammals, as well as pharmaceutically acceptable biological precursors (otherwise known as prodrugs) of the compounds of formula (I), are also within the scope of the invention.
In addition to the above, as known to those skilled in the art, the unsubstituted nitrogen on the pyrazole ring of the compounds of formula (I) equilibrates rapidly in solution to form a mixture of tautomers, as described below:
wherein X, Ar, R1, R2 and R3 are as defined above.
Thus, in the present invention, where only one tautomer of the compound of formula (I) is represented, the other tautomer (Ia) is also within the scope of the invention, unless explicitly noted otherwise.
Unless otherwise indicated, general terms used herein have the meanings reported below.
The term "straight or branched C1-C6Alkyl "means saturated aliphatic hydrocarbon groups including straight and branched chain groups of 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituents are preferably 1-3, independently selected from halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted C3-C6Cycloalkyl radicalsHeterocyclyl and aryl, wherein R4, R5, R6, R7, R8, R9 and R10 are as defined above.
The term "C3-C6Cycloalkyl "means a 3-to 6-membered all carbon monocyclic ring which may contain one or more double bonds but which does not have a completely conjugated pi-electron system. Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl and cyclohexadienyl, but are not limited thereto. Cycloalkyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably 1 or 2 substituents independently selected from halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl and aryl, wherein R4, R5, R6, R7, R8, R9 and R10 are as defined above.
The term "heterocyclyl" means a 3-to 7-membered saturated or partially unsaturated carbocyclic ring in which one or more carbon atoms are replaced by heteroatoms, such as nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclic groups are, for example, oxiranyl, aziridinyl, glycidylalkyl, azetidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, pyranyl, dihydropyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, pyrazolinyl, isoxazolyl, and the likeOxazolidinyl, isoOxazolinyl, thiazolidinyl, thiazolinyl, isothiazolinyl, and isothiazolinylAlkyl radicalPiperazinyl, morpholinyl, thiomorpholinyl, hexamethyleneimino (exomethyleneimino), homopiperazinyl, and the like. The heterocyclic group may be substituted or unsubstituted. When substituted, the substituents are preferably 1 or 2 substituents independently selected from halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl and aryl, wherein R4, R5, R6, R7, R8, R9 and R10 are as defined above.
The term "aryl" means a mono-, bi-or multi-carbocyclic hydrocarbon having from 1 to 4 ring systems, wherein the individual rings are optionally further fused or connected to each other by single bonds, wherein at least one carbocyclic ring is "aromatic", wherein the term "aromatic" means a completely conjugated pi-electron bond system. Non-limiting examples of such aryl groups are phenyl, alpha-or beta-naphthyl or biphenyl.
The term "heteroaryl" means an aromatic heterocycle, typically a 5-to 7-membered heterocycle having 1-3 heteroatoms selected from N, O or S; the heterocyclic ring may optionally be further fused or linked to aromatic and non-aromatic carbocyclic and heterocyclic rings. Non-limiting examples of such heteroaryl groups are, for example, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, imidazolyl, thiazolyl, isothiazolyl, pyrrolyl, phenyl-pyrrolyl, furyl, phenyl-furyl,azolyl radical, isoAzolyl, pyrazolyl, thienyl, benzothienyl, isoindolinyl, benzimidazolyl, quinolyl, isoquinolyl, 1,2, 3-triazolyl, 1-phenyl-1, 2, 3-triazolyl, 2, 3-indolinyl, 2, 3-dihydrobenzofuranA group, 2, 3-dihydrobenzothienyl; benzopyranyl, 2, 3-dihydrophenylpropylOxazinyl, 2, 3-dihydroquinoxalinyl, and the like.
Aryl and heteroaryl groups may be optionally substituted by one or more, preferably 1,2 or 3 substituents independently selected from halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substituted, linear or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl and aryl, wherein R4, R5, R6, R7, R8, R9 and R10 are as defined above.
The term "halogen" denotes fluorine, chlorine, bromine or iodine.
The term "C2-C6Alkenyl "denotes an aliphatic C group which contains at least one carbon-carbon double bond and which may be straight-chain or branched2-C6A hydrocarbon chain. Representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-or 2-butenyl, and the like.
The term "C2-C6Alkynyl "denotes an aliphatic C group containing at least one carbon-carbon triple bond and which may be straight or branched2-C6A hydrocarbon chain. Representative examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-or 2-butynyl, and the like.
The term "cyano" denotes a-CN residue.
The term "nitro" denotes-NO2A group.
The term "pharmaceutically acceptable salts" of the compounds of formula (I) refers to those salts which retain the biological effectiveness and properties of the parent compound. Such salts include those intended to retain the biological effectiveness and properties of the parent compound. Examples of such salts include: acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid and the like, or with organic acids such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, lactic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, benzoic acid, p-toluenesulfonic acid, salicylic acid, cinnamic acid, mandelic acid, tartaric acid, citric acid, succinic acid, malonic acid and the like; salts formed when the acidic proton present in the compound of formula (I) is replaced by a metal ion, such as an alkali metal ion, for example sodium or potassium, or an alkaline earth metal ion, for example calcium or magnesium, or is coordinated with an organic base, such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.
The compound of formula (I) is represented by general formula (I)A) Wherein X is-CH2-：
The compound of formula (I) is represented by general formula (I)B) Wherein X is-CH (OH) -:
the compound of formula (I) is represented by general formula (I)C) Wherein X is-CH (OR') -:
the compound of formula (I) is represented by general formula (I)D) Wherein X is-C (R ' R ' ') -:
a preferred class of compounds of formula (I) is the following compounds, wherein:
x is-CH2-, -CH (OH) -, -CH (OR ') -, OR-C (R' R ') -, in which R' is C1-C3Alkyl and R' is hydrogen or C1-C3An alkyl group;
r is optionally substituted C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, and
r1, R2 and R3 are independently hydrogen, halogen or hydroxy.
Another preferred type of compound of formula (I) is the following compound, wherein:
x is-CH2-, -CH (OH) -, -CH (OR ') -, OR-C (R ' R ') -, wherein R ' is methyl and R ' is hydrogen OR methyl, and
r1, R2 and R3 are hydrogen.
Another preferred type of compound of formula (I) is the following compound, wherein:
r is optionally substituted aryl or heteroaryl.
A more preferred class of compounds of formula (I) is the following compounds, wherein:
ar is a group of the formula:
wherein Ra, Rb and Rc are independently hydrogen, halogen, C2-C6Alkenyl radical, C2-C6Alkynyl, cyano, nitro, NHCOR4, COR4, NR5R6, NR5COR4, OR7, SR7, SOR10, SO2R10，NHSOR10，NHSO2R10，R8R9N-C1-C6Alkyl radical, R8O-C1-C6Alkyl, optionally further substitutedStraight or branched C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein R4, R5, R6, R7, R8, R9 and R10 are as defined above, and
r is an optionally substituted aryl group.
Another more preferred type of compound of formula (I) is the following compound, wherein:
ar is a group of the formula:
wherein Ra and Rb are as defined above.
The most preferred type of compound of formula (I) is the following compound, wherein:
ar is a group of the formula:
wherein Ra is hydrogen, halogen, nitro, NHCOR4 OR NR5R6, and Rb is hydrogen, nitro, NR5R6, OR7 OR R8R9N-C1-C6Alkyl groups, wherein R4, R5, R6, R7, R8 and R9 are as defined above.
Specific compounds of the invention (cpd.) are listed below:
n- (5-benzyl-1H-indazol-3-yl) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (2, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitrobenzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide;
2-amino-N- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- [5- (2, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (1-methyl-piperidin-4-ylamino) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (1-methyl-piperidin-4-ylamino) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide:
n- [5- (2, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
2-cyclohexylamino-N- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
2-cyclohexylamino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (4-hydroxy-cyclohexylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (4-hydroxy-cyclohexylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2-isobutylamino-4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-isobutylamino-4- (4-methyl-piperazin-1-yl) -benzamide;
2-benzylamino-N- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
2-benzylamino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- ((S) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- ((S) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- ((R) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- ((R) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2- (3-fluoro-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- (3-fluoro-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-phenylamino-benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-phenylamino-benzamide;
43.1H-pyrrole-2-carboxylic acid [2- [5- (3-fluoro-benzyl) -1H-indazol-3-ylcarbamoyl ] -5- (4-methyl-piperazin-1-yl) -phenyl ] -amide;
44.1H-pyrrole-2-carboxylic acid [2- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylcarbamoyl ] -5- (4-methyl-piperazin-1-yl) -phenyl ] -amide;
45.1H-pyrrole-3-carboxylic acid [2- [5- (3-fluoro-benzyl) -1H-indazol-3-ylcarbamoyl ] -5- (4-methyl-piperazin-1-yl) -phenyl ] -amide;
46.1H-pyrrole-3-carboxylic acid [2- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylcarbamoyl ] -5- (4-methyl-piperazin-1-yl) -phenyl ] -amide;
n- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -2-methanesulfonylamino-4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-methanesulfonylamino-4- (4-methyl-piperazin-1-yl) -benzamide;
49.2-fluoro-N- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -5- (tetrahydro-pyran-4-ylamino) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5- (tetrahydro-pyran-4-ylamino) -benzamide;
51.2-fluoro-N- [5- (3-fluoro-benzyl) -1H-indazol-3-yl ] -5- (2-methoxy-ethylamino) -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5- (2-methoxy-ethylamino) -benzamide;
4- [ (3-dimethylamino-propyl) -methyl-amino ] -N- [5- (3-ethoxy-benzyl) -1H-indazol-3-yl ] -2-nitro-benzamide;
54.2-amino-4- [ (3-dimethylamino-propyl) -methyl-amino ] -N- [5- (3-ethoxy-benzyl) -1H-indazol-3-yl ] -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (3-dimethylamino-propyl) -methyl-amino ] -2- (tetrahydro-pyran-4-ylamino) -benzamide;
56.N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (3-dimethylamino-propyl) -methyl-amino ] -2- (2-methoxy-1-methoxymethyl-ethylamino) -benzamide;
57.2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (3-dimethylamino-propyl) -methyl-amino ] -benzamide;
58.N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (3-dimethylamino-propyl) -methyl-amino ] -benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (3-dimethylamino-propyl) -methyl-amino ] -2-nitro-benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
61.N- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -benzamide;
63.N- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
65.N- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
66.N- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3-ethoxy-phenyl) -hydroxy-methyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide;
68.N- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -benzamide;
71.N- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
73.N- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl ] -1H-indazol-3-yl } -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [1- (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
76.N- {5- [1- (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
77.N- {5- [ (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -benzamide;
78.N- {5- [ (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
79.N- {5- [ (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
80.N- {5- [ (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [ (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
82.N- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -benzamide;
85.N- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
86.N- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -2- (2-methoxy-1, 1-dimethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
88.N- {5- [1- (3, 5-difluoro-phenyl) -1-methyl-ethyl ] -1H-indazol-3-yl } -2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
89.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methyl-1, 4-diazepan-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
90.N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- (tetrahydro-pyran-4-ylamino) -benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [4- (dimethylamino) piperidin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
92.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
93.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -3- (4-methylpiperazin-1-yl) benzamide;
94.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2S) -1-methylpyrrolidin-2-yl ] methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
95.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (1-methylpiperidin-4-yl) oxy ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
96.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [2- (dimethylamino) ethoxy ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (3S) -1-methylpyrrolidin-3-yl ] oxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
98.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (piperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
99.N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methylpiperazin-1-yl) -2- { [ cis-4- (trifluoromethyl) cyclohexyl ] amino } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methylpiperazin-1-yl) -2- { [ trans-4- (trifluoromethyl) cyclohexyl ] amino } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-fluoro-4- (4-methylpiperazin-1-yl) benzamide 101;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -1- (piperidin-4-yl) -1H-pyrazole-4-carboxamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- [ (cis-4-hydroxycyclohexyl) amino ] -4- (4-methylpiperazin-1-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- [ (trans-4-hydroxycyclohexyl) amino ] -4- (4-methylpiperazin-1-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- [ (2-hydroxyethyl) amino ] -4- (4-methylpiperazin-1-yl) benzamide;
106.2- [ (azetidin-3-ylmethyl) amino ] -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methylpiperazin-1-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- { [ (1-methylazetidin-3-yl) methyl ] amino } -4- (4-methylpiperazin-1-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (1-methylpiperidin-4-yl) amino ] -2- [ tetrahydro-2H-pyran-4-ylamino ] benzamide;
109.4- [ (azetidin-3-ylmethyl) amino ] -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- [ (1-methylpiperidin-4-yl) amino ] benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- [ (1-methylpiperidin-4-yl) amino ] -4- (morpholin-4-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-methoxy-4- (4-methylpiperazin-1-yl) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -5- (4-methylpiperazin-1-yl) -3- (tetrahydro-2H-pyran-4-ylamino) pyridine-2-carboxamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -6- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) pyridine-3-carboxamide;
115.1- [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) benzyl ] piperidine;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2-methoxyethyl) (methyl) amino ] methyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (pyrrolidin-1-ylmethyl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (morpholin-4-ylmethyl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
119.4- (azetidin-1-ylmethyl) -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5- (4-methyl-piperazin-1-ylmethyl) -benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-fluoro-5- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] methyl } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-fluoro-5- (morpholin-4-ylmethyl) benzamide;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5- ((S) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [4- (pyrrolidin-1-yl) piperidin-1-yl ] carbonyl } benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
129.N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]-N4-methyl-2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [4- (propan-2-yl) piperazin-1-yl ] carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
131.N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]-2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (4-methylpiperazin-1-yl) carbonyl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [4- (dimethylamino) piperidin-1-yl ] carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
134.N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- (1-methylpiperidin-4-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide;
n- [5- (2-methyl-5-fluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
4- (4-methylpiperazin-1-yl) -N- [5- (pyridin-3-ylmethyl) -1H-indazol-3-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [ 5-benzyl-1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide;
138.4- { [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl ] amino } piperidine-1-carboxylic acid ethyl ester;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methylpiperazin-1-yl) -2- (piperidin-4-ylamino) benzamide;
ethyl 5- (3, 5-difluorobenzyl) -3- ({ [4- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl ] carbonyl } amino) -1H-indazole-1-carboxylate;
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2- ((S) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2R) -1-methylpyrrolidin-2-yl ] methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (3R) -1-methylpyrrolidin-3-yl ] oxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide;
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-fluoro-5- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] methyl } benzamide, and
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5- ((R) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -benzamide.
Preferred specific compounds of the invention are:
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide.
The present invention also provides a process for the preparation of a compound of formula (I) as defined above, characterised in that it comprises:
i) reducing a carbonyl compound of formula (II):
wherein Ar, R, R1, R2, and R3 are as defined above, to give formula (I)A)，(IB) Or (I)C) The compound of (1):
wherein Ar, R, R1, R2, R3 and R' are as defined above;
or
i') reaction of formula (III)A)，(IIIB)，(IIIC) Or (III)D) The compound of (1):
wherein R, R1, R2, R3, R ' and R ' ' are as defined above, with a compound of formula (IV):
wherein Ar is as defined above and Y represents hydroxy or a suitable leaving group, for example halogen, to give a compound of formula (I) as defined above;
or
i') formula (XXII)A)，(XXIIC) Or (XXII)D) Deprotection of the compound of (1):
wherein Ar, R, R1, R2, R3, R ' and R ' ' are as defined above and PG is a suitable protecting group, such as benzyl, p-methoxybenzyl, o, p-dimethoxybenzyl or triphenylmethyl, to give the formula (I)A)，(IC) Or (I)D) The compound of (1):
wherein Ar, R1, R2, R3, R' and R "are as defined above, optionally separating the resulting compound into single isomers, converting the compound of formula (I) into a different compound of formula (I), and/or if desired into a pharmaceutically acceptable salt.
The invention further provides a process for the preparation of formula (I) as defined aboveA),(IB) Or (I)C) Characterized in that a compound of formula (II) as defined above is prepared according to the following steps:
a) reacting a compound of formula (XII):
wherein R1, R2 and R3 are as defined above, with an organometallic compound of the formula RMgZ (XIII) (i.e. a Grignard reagent) wherein R is as defined above and Z is halogen, to give a compound of the formula (XI):
wherein R, R1, R2 and R3 are as defined above;
b) oxidizing the resulting compound of formula (XI) to obtain a compound of formula (X):
wherein R, R1, R2 and R3 are as defined above;
c) reacting the resulting compound of formula (X) with hydrazine hydrate to give a compound of formula (IX):
wherein R, R1, R2 and R3 are as defined above;
d) protecting the resulting compound of formula (IX) to give a compound of formula (VIII):
wherein R, R1, R2 and R3 are as defined above, and PG1Are suitable protecting groups, such as trifluoroacetyl;
e) protecting the resulting compound of formula (VIII) to yield a compound of formula (VII):
wherein R, R1, R2, R3, PG and PG1As defined above;
f) removing the protecting group PG from the resulting compound of formula (VII)1To give a compound of formula (VI):
wherein R, R1, R2, R3 and PG are as defined above;
g) reacting the compound of formula (VI) obtained with a compound of formula (IV) as defined above to obtain a compound of formula (V):
wherein Ar, R, R1, R2, R3 and PG are as defined above;
h) the compound of formula (V) obtained is then subjected to a de-inclusion to obtain the compound of formula (II) as defined above.
The invention further provides a process for the preparation of formula (I) as defined aboveA) Characterized in that the compound of formula (III) as defined above is prepared according to the following procedureA) The compound of (1):
j) in appropriate reagents, e.g. NaI and Me3Reducing a compound of formula (XI) as defined above in the presence of SiCl to give a compound of formula (XIV):
wherein R, R1, R2 and R3 are as defined above;
or
k) Reacting a boronic acid compound of formula (XV):
wherein R1, R2 and R3 are as defined above, with a compound of formula (XVI):
wherein R is as defined above and W represents a halogen atom, for example bromine or iodine, or a suitable leaving group, such as a sulfonate, for example a mesylate or triflate, or a phosphate ester, in the presence of a suitable catalyst, for example a palladium catalyst, to give a compound of formula (XIV) as defined above;
l) reacting the compound of formula (XIV) obtained with hydrazine hydrate to obtain the compound of formula (III) as defined aboveA) The compound of (1).
The invention further provides a process for the preparation of formula (I) as defined aboveB) Characterized in that the compound of formula (III) as defined above is prepared according to the following procedureB) The compound of (1):
l') reacting a compound of formula (XI) as defined above with hydrazine hydrate to give a compound of formula (III) as defined aboveB) Is/are as followsA compound is provided.
The invention further provides a process for the preparation of formula (I) as defined aboveC) Characterized in that the compound of formula (III) as defined above is prepared according to the following procedureC) The compound of (1):
m) reacting a compound of formula (XI) as defined above with an electrophilic alkylating agent of formula (XVIII):
R'-W'
(XVIII)
wherein R 'is as defined above and W' represents a halogen atom, for example chlorine, bromine or iodine, or a suitable leaving group, such as a sulfonate, for example mesylate or triflate, to give a compound of formula (XVII):
wherein R, R1, R2, R3 and R' are as defined above;
l '') reacting the compound of formula (XVII) obtained with hydrazine hydrate to obtain the compound of formula (III) as defined aboveC) The compound of (1).
The invention further provides a process for the preparation of formula (I) as defined aboveD) Characterized in that the compound of formula (III) is prepared according to the following procedureD1) Wherein R' is hydrogen:
wherein R, R1, R2, R3 and R' are as defined above:
n) reacting a compound of formula (XIV) as defined above with a compound of formula (XVIII) as defined above;
l ' ' ') the resulting compound of formula (XIX)D1) Transformation ofCompound (a):
wherein R, R1, R2, R3 and R' are as defined above, with hydrazine hydrate to give formula (III) as defined aboveD1) A compound of (1);
or
o) reacting a compound of formula (XXI):
wherein R1, R2, R3 and R' are as defined above, with a compound of formula (XIII) as defined above to give a compound of formula (XX):
wherein R, R1, R2, R3 and R' are as defined above;
p) reduction of the resulting compound of formula (XX) to give the compound of formula XIX as defined aboveD1The compound of (1).
The invention further provides a process for the preparation of formula (I) as defined aboveD) Characterized in that a compound of formula (III) having the following formula is prepared according to the following procedureD2) Wherein R "is as defined above, but is not hydrogen:
wherein R, R1, R2, R3 and R' are as defined above:
q) is as defined aboveFormula (XIX)D1) With an electrophilic alkylating agent of formula (XXIII):
R''-W'
(XXIII)
wherein R 'and W' are as defined above, to give formula (XIX)D2) The compound of (1):
wherein R, R1, R2, R3 and R' are as defined above and R "is as defined above, but is not hydrogen;
liv) The obtained formula (XIX)D2) With hydrazine hydrate to give the compound of formula (III) as defined aboveD2) The compound of (1).
The invention further provides a process for the preparation of formula (I) as defined aboveA),(IC) Or (I)D) Characterized in that a compound of the formula (XXII) as defined above is prepared according to the following procedureA)，(XXIIC) Or (XXII)D) The compound of (1):
r) protection of formula (III) as defined aboveA)，(IIIC) Or (III)D) To give a compound of the formula (XXIV)A)，(XXIVC) Or (XXIV)D) The compound of (1):
wherein R, R1, R2, R3, R, R '' and PG1As defined above;
s) protected by a compound of formula (XXIV)A)，(XXIVC) Or (XXIV)D) To give a compound of the formula (XXV)A)，(XXVC) Or (XXV)D) The compound of (1):
wherein R, R1, R2, R3, R, R '', PG and PG1As defined above;
t) from the resulting compound of formula (XXV)A)，(XXVC) Or (XXV)D) Removing the protecting group PG from the compound of (1)1Obtaining the formula (XXVI)A)，(XXVIC) Or (XXVI)D) The compound of (1):
wherein R, R1, R2, R3, R, R '' and PG are as defined above;
u) the resulting compound of formula (XXVI)A)，(XXVIC) Or (XXVI)D) With a compound of formula (IV) as defined above, to give a compound of formula (XXII) as defined aboveA)，(XXIIC) Or (XXII)D) The compound of (1).
Note that the compound of formula (V) as defined above may be in either of its isomeric forms a or b or a mixture of both:
likewise, formula (XXII) as defined aboveA)，(XXIIC)，(XXIID)，(XXVA)，(XXVC)，(XXVD)，(XXVIA)，(XXVIC) And (XXVI)D) The compound of (b) may be in any of its isomeric forms a or b.
The compounds of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) To another compound of formula (II),(V)，(XXIIA)，(XXIIC) And (XXII)D) The conversion being carried out by one or more of the following reactions:
1) reduction of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein Ar is substituted aryl and one of the substituents is NO2To give compounds of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein the substituent is NH2；
2) Acylated (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein Ar is substituted aryl and one of the substituents is NH2By reaction with an acylating agent of the formula (XXVII) or (XXVIII):
wherein R4 and Y are as defined above, to give compounds of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein the substituent is NHCOR4 or NHSO2R4 residue, wherein R4 is as defined above;
3) the compounds of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein Ar is substituted aryl and one of the substituents is NH2With the appropriate aldehyde or ketone in the presence of a reducing agent to give compounds of the formulae (II), (V), (XXII)A)，(XXIIC) And (XXII)D) Wherein the substituent is a NR5R6 group, wherein one of R5 or R6 is hydrogen and the other is optionally further substituted straight or branched chain C1-C6Alkyl radical, C3-C6Cycloalkyl, heterocyclyl, aryl, R8R9N-C2-C6Alkyl radical, R8O-C2-C6Alkyl, wherein R8 and R9 are as defined above.
A compound of formula (I) may be converted to another compound of formula (I) by one or more of the following reactions:
4) reduction of a compound of formula (I) wherein Ar is substituted aryl and one of the substituents is NO2To give a compound of formula (I) wherein the substituent is NH2；
5) Acylating a compound of formula (I) wherein Ar is substituted aryl and one of the substituents is NH2By reaction with a compound of formula (XXVII) or (XXVIII) as defined above, followed by selective deprotection of the acyl group on the pyrazole ring, to give a compound of formula (I) wherein the substituent is NHCOR4 or NHSO2R4 residue, wherein R4 is as defined above;
6) reacting a compound of formula (I) wherein Ar is substituted aryl and one of the substituents is NH2With a suitable aldehyde or ketone in the presence of a reducing agent to give a compound of formula (I) wherein the substituent is a NR5R6 group wherein one of R5 or R6 is as defined in conversion 3).
The synthesis of the compounds of formula (I) may be carried out in a stepwise manner according to the synthetic procedures described above, whereby each intermediate is isolated and purified by standard purification techniques, such as column chromatography, before carrying out the subsequent reaction. Alternatively, two or more steps of the synthetic sequence may be performed in a so-called "one-shot" operation, as known in the art, whereby only the compounds resulting from the two or more steps are isolated and purified.
Schemes 1-4 shown below represent the preparation of compounds of formula (I) wherein X, Ar, R1, R2 and R3 have the above-described meanings.
Scheme 1
Scheme 2
Scheme 3
Scheme 4
According to step I), the compound of formula (II) may be reduced by various means and experimental conditions well known in the art to give formula (I)A)，(IB) Or (I)C) The compound of (1). The above reduction is preferably carried out in the presence of sodium borohydride, sodium cyanoborohydride, sodium borohydride/trifluoroacetic acid, zinc/hydrochloric acid, tin chloride/acetic acid in a suitable solvent such as toluene, dichloromethane, chloroform, diethyl ether, tetrahydrofuran, 1, 4-bisAlkane, methanol, ethanol, isopropanol, acetic acid, reaction temperature from about-10 ℃ to reflux temperature, and reaction time varied from about 1 hour to about 96 hours. Depending on the experimental conditions, the formula (I)A)，(IB) Or (I)C) The compound of (a) is isolated as the main product.
According to step i'), the compound of formula (III)A)，(IIIB)，(IIIC) Or (III)D) With a compound of formula (IV) according to various means and experimental conditions for condensation reactions widely known in the art to give compounds of formula (I)A)，(IB)，(IC) Or (I)D) The compound of (1). The compound of formula (IV) wherein Y is hydroxy is preferably converted to its corresponding compound in the presence of thionyl chloride or oxalyl chloride in a suitable solventWherein Y is chlorine, and suitable solvents are, for example, toluene, dichloromethane, chloroform, diethyl ether, tetrahydrofuran, 1, 4-bisAlkane, reaction temperature from about-10 ℃ to reflux temperature, reaction time varying between about 1 hour to about 96 hours. Isolation of the acid chloride by evaporation of the solvent and further reaction with (III)A)，(IIIB)，(IIIC) Or (III)D) In the presence of a base such as pyridine, triethylamine or N-ethyldiisopropylamine in a suitable solvent such as toluene, dichloromethane, chloroform, diethyl ether, tetrahydrofuran, 1, 4-bis (methylene chloride)Alkane, at a reaction temperature of from about-40 ℃ to reflux temperature, for a reaction time of from about 1 hour to about 96 hours. Alternatively, reacting a compound of formula (IV) with a compound of formula (III)A)，(IIIB)，(IIIC) Or (III)D) In the presence of an activating agent such as hydroxybenzotriazole, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-ethyl-3- (3' -dimethylamino) carbodiimide hydrochloride. Preferably, the reaction is carried out in a suitable solvent, such as tetrahydrofuran, dichloromethane, toluene, 1, 4-bisAn alkane and the proton scavenger is, for example, pyridine, triethylamine, N-diisopropylethylamine, the reaction temperature is from about room temperature to reflux, and the reaction time is from about 30 minutes to about 96 hours.
According to step i ″), formula (XXII) can be reacted by various means and experimental conditions widely known in the artA)，(XXIIC) Or (XXII)D) Deprotection of the compound of formula (I) to giveA)，(IC) Or (I)D) The compound of (1). Preferably, for acyl residues, the reaction is carried out under basic conditions, e.g., in the presence of sodium hydroxide, potassium hydroxide, sodium hydroxide,Lithium or barium hydroxide or a tertiary amine, such as triethylamine or diisopropylethylamine or hydrazine, in a suitable solvent, such as methanol, ethanol, tetrahydrofuran, N, N-dimethylformamide, water and mixtures thereof. Generally, the reaction is carried out at room temperature to reflux temperature for a time ranging from about 30 minutes to about 96 hours. Where PG represents a suitable protecting group, for example benzyl, p-methoxybenzyl, o, p-dimethoxybenzyl or triphenylmethyl, the conversion may be carried out under the same conditions as reported in step h).
According to step a), the compound of formula (XII) can be converted into the compound of formula (XI) according to conventional methods widely known in the literature by using a grignard reagent of formula (XIII) in various ways and experimental conditions. Preferably, the compound of formula (XII) is reacted with an organometallic reagent in a suitable solvent, e.g., tetrahydrofuran, 1, 4-bisIn an alkane and diethyl ether at a temperature of-78 ℃ to room temperature for about 30 minutes to 96 hours.
According to step b), the compound of formula (XI) can be oxidized to the compound of formula (X) in various ways according to conventional methods for oxidizing alcohol compounds to ketone compounds. Preferably, the reaction is carried out in a suitable solvent, such as methanol, ethanol, t-butanol, water, tetrahydrofuran, 1, 4-bisAlkanes, toluene, acetic acid, trifluoroacetic acid, dichloromethane, dichloroethane, acetonitrile, dimethyl sulfoxide or mixtures thereof, such as 3-chloroperbenzoic acid, hydrogen peroxide, Dess-Martin oxidizer (Dess-Martin periodinane), ozone, potassium permanganate, sodium periodate, periodic acid, and the catalysts are chromium (VI) oxide, tetrapropylammonium perruthenate, ruthenium chloride. Generally, the reaction is carried out at-78 ℃ to reflux temperature for about 30 minutes to about 96 hours.
According to step c), the compound of formula (X) can be converted into the compound of formula (IX) by various means and experimental conditions widely known in the art for the preparation of 3-aminoindazoles. Preferably, the reaction of the compound of formula (X) with hydrazine is carried out in a suitable solvent, e.g. toluene, tetrahydrofuran, 1, 4-bisThe alkane, dimethylsulfoxide, acetonitrile, methanol, ethanol or n-butanol neutralization is carried out at 0 ℃ to reflux temperature for about 1 hour to about 96 hours. To catalyze the reaction, it may be necessary to add an acid, for example, preferably hydrochloric acid or acetic acid.
According to step d), the compound of formula (IX) can be converted into the compound of formula (VIII) by various means and experimental conditions for protecting primary amino groups, which are widely known in the art. The reaction is preferably carried out by treatment with an excess of trifluoroacetic anhydride or trifluoroacetyl chloride in a suitable solvent, for example acetonitrile, tetrahydrofuran, toluene, dichloromethane. Generally, the reaction is carried out at a temperature of from 0 ℃ to about 110 ℃ for from about 30 minutes to about 96 hours. Work-up of the reaction mixture with a protic solvent, for example, water, methanol, ethanol or mixtures thereof or with aqueous sodium bicarbonate results in selective hydrolysis of the trifluoroacetyl group on the indazole ring. For the preparation of phthalimide derivatives, the reaction is carried out by treatment with phthalic anhydride under basic conditions, for example in the presence of 1, 8-diazabicyclo [5.4.0] undec-7-ene, N-dimethylaminopyridine, pyridine, triethylamine and in a suitable solvent, for example acetonitrile, tetrahydrofuran, N-dimethylformamide, toluene, dichloromethane, water and mixtures thereof. Generally, the reaction is carried out at a temperature of from room temperature to about 110 ℃ for from about 30 minutes to about 96 hours.
The reaction according to step e) to obtain the compound of formula (VII) from the compound of formula (VIII) can be carried out in various ways and under experimental conditions. When PG is a triphenylmethyl group, the reaction is preferably carried out by treatment with a trityl chloride in a suitable solvent and in the presence of a proton scavenger, said reaction being carried outSolvents such as tetrahydrofuran, dichloromethane, toluene, 1, 4-bisAn alkane, preferably a proton scavenger such as 1, 8-diazabicyclo [5.4.0]]Undec-7-ene, triethylamine, N, N-diisopropylethylamine, pyridine, reaction temperature from room temperature to reflux temperature, reaction time from about 30 minutes to about 96 hours.
According to step f), the compound of formula (VII) can be converted into the compound of formula (VI) according to conventional methods by removing the appropriate protecting group, e.g. trifluoroacetyl group. The reaction is preferably carried out by treatment with an organic or inorganic base such as potassium carbonate, sodium hydroxide, ammonia, triethylamine, N, N-diisopropylethylamine in a suitable solvent such as tetrahydrofuran, dichloromethane, toluene, 1, 4-diisopropylethylamineAlkane, methanol, ethanol, water or mixtures thereof. The reaction temperature is between room temperature and reflux temperature, and the reaction time is between about 30 minutes and about 96 hours.
According to step g), the compound of formula (VI) may be converted to the compound of formula (V) in various ways and experimental conditions widely known in the art for condensation reactions. Preferably the reaction is carried out in the same manner as reported in step i').
According to step h), the compound of formula (V) can be converted to the compound of formula (II) by deprotecting the endocyclic indazole nitrogen atom according to conventional procedures capable of selectively hydrolysing benzyl, 4-methoxybenzyl, 2, 4-dimethoxybenzyl and triphenylmethyl protecting groups. Preferably the reaction is carried out under acidic conditions, preferably in the presence of an inorganic or organic acid, such as hydrochloric acid, trifluoroacetic acid or methanesulfonic acid, and in a suitable solvent, such as dichloromethane, 1, 4-bisAlkanes, lower alcohols (e.g. methanol orEthanol) at a temperature of from room temperature to about 80 deg.c for a time of from about 1 hour to about 48 hours. In the alternative, the reaction is carried out under reducing conditions, for example in the presence of hydrogen and a hydrogenation catalyst, in a suitable solvent, for example ethanol, methanol, ethyl acetate or mixtures thereof. The catalyst is typically a metal, most commonly a palladium derivative, such as palladium hydroxide or palladium black.
According to step j), the compound of formula (XI) can be reduced to the compound of formula (XIV) in various ways according to conventional methods for reducing alcohol compounds to alkanes. Preferably, the reaction is carried out in a suitable solvent, such as methanol, ethanol, tetrahydrofuran, 1, 4-bis, and in the presence of a suitable reducing systemAlkanes, acetic acid, dichloromethane, acetonitrile or mixtures thereof, such as trimethylsilyl chloride/sodium iodide, dichlorodimethylsilane/sodium iodide, triethylsilane/trifluoroacetic anhydride, sodium borohydride/trifluoroacetic acid. Generally, the reaction is carried out at a temperature of-10 ℃ to reflux for about 30 minutes to about 96 hours.
According to step k), the compound of formula (XV) can be converted in various ways into the compound of formula (XIV) according to conventional methods for boron-derivative coupling, i.e. Suzuki-like reactions, in the presence of the compound of formula (XVI). Preferably, the reaction is carried out in a suitable solvent, such as ethanol, water, tetrahydrofuran, bisAn alkane, acetone, N-dimethylformamide, dimethoxyethane, toluene, xylene or a mixture thereof, such as triethylamine, diisopropylethylamine, sodium carbonate, potassium carbonate or cesium carbonate, potassium phosphate, sodium hydroxide or cesium fluoride, at a reaction temperature of-20 ℃ to reflux temperature for a reaction time of about 1 hour to about 96 hours. The catalyst is generally in the presence of a suitable ligand, for example triphenylphosphineThe following metals, most commonly palladium derivatives, such as palladium chloride, palladium acetate.
According to step l), the compound of formula (XIV) can be converted into formula (III) in various ways and experimental conditionsA) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step c).
The compounds of the formula (XI) can be converted into the formula (III) according to step l') in various ways and under experimental conditionsB) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step c).
According to step m), the compound of formula (XI) can be converted in various ways into the compound of formula (XVII) in the presence of the compound of formula (XVIII) according to conventional methods for O-alkylation reactions. Preferably, the reaction is carried out in a suitable solvent, such as tetrahydrofuran, diethyl ether and the like, and in the presence of a suitable baseAlkane, N-dimethylformamide, dimethoxyethane, said base being, for example, triethylamine, diisopropylethylamine, sodium carbonate, potassium carbonate or cesium carbonate, sodium hydride, at a reaction temperature of-78 ℃ to reflux temperature, for a reaction time of about 1 hour to about 96 hours. Alkylating agents are typically halogen or sulfonate derivatives; the most common leaving group is iodine, bromine, triflate, or mesylate.
According to step l "), the compounds of formula (XVII) can be converted into formula (III) in various ways and experimental conditionsC) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step c).
According to step n), the compound of formula (XIV) can be converted into the compound of formula (XIX) in various ways according to conventional methods for C-alkylation reactions in the presence of the compound of formula (XVIII)D1) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step m).
According to step l ' ' ') can be variedMode for the invention and Experimental conditionsD1) Is converted into the compound of formula (III)D1) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step c).
According to step o), a compound of formula (XXI) can be converted into a compound of formula (XX) in the presence of a compound of formula (XIII) in various ways and experimental conditions. Preferably the reaction is carried out in the same manner as reported for step a).
According to step p), the compound of formula (XX) may be converted into formula (XIX) in various ways and experimental conditionsD1) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step j).
According to step q), the compound of the formula (XIX) can be reacted in various ways and under experimental conditions in the presence of a compound of the formula (XXIII)D1) Is converted into the compound of formula (XIX)D2) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step m).
According to step lIV) The formula (XIX) can be used in various ways and under experimental conditionsD2) Into the compound of formula (III)D2) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step c).
According to step r), formula (III) can be reacted in various ways and experimental conditions for protecting primary amino groups, which are widely known in the artA)，(IIIC) Or (III)D) Is converted to the compound of formula (XXIV)A)，(XXIVC) Or (XXIV)D) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step d).
According to step s), the reaction of formula (XXIV) can be carried out in various ways and under experimental conditionsA)，(XXIVC) Or (XXIV)D) To give the compound of formula (XXV)A)，(XXVC) Or (XXV)D) Reaction of the compound (2). Preferably the reaction is carried out in the same manner as reported for step e).
According to step t), the compound of formula (XXV) can be prepared by removing the appropriate protecting group, e.g. trifluoroacetyl group, according to conventional methodsA)，(XXVC) Or (XXV)D) Is converted to the compound of formula (XXVI)A)，(XXVIC) Or (XXVI)D) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step f).
According to step u), formula (XXVI) can be used in various ways and experimental conditions for the condensation reaction widely known in the artA)，(XXVIC) Or (XXVI)D) Is converted to the compound of formula (XXII)A)，(XXIIC) Or (XXII)D) The compound of (1). Preferably the reaction is carried out in the same manner as reported for step i').
According to the transformation described in 1), in various ways, the compounds of the formulae (II), (V), (XXII) are reacted according to conventional methods well known in the literatureA)，(XXIIC) Or (XXII)D) Wherein Ar is substituted aryl and one of the substituents is nitro, to the compound of formula (II), (V), (XXII)A)，(XXIIC) Or (XXII)D) Wherein the substituent is amino. Preferably, the conversion is carried out in a suitable solvent, such as methanol, ethanol, water, tetrahydrofuran, 1, 4-bisAlkane, N-dimethylformamide, acetic acid or mixtures thereof, the reducing agent being, for example, hydrogen and a hydrogenation catalyst, or the conversion being carried out by treatment with cyclohexene or cyclohexadiene or formic acid or ammonium formate and a hydrogenation catalyst or a metal, for example iron or zinc, in the presence of a mineral acid, for example hydrochloric acid, or by treatment with tin (II) chloride, the reaction temperature being from 0 ℃ to reflux temperature, the reaction time being from about 1 hour to about 96 hours. The hydrogenation catalyst is typically a metal, most commonly palladium, which may be used directly or supported on carbon.
According to the transformation described in 2), in various ways, the rootsAcylation of the formula (II), (V), (XXII) by reaction with an acetylating agent of the formula (XXVII) or (XXVIII) according to conventional methods well known in the literatureA)，(XXIIC) Or (XXII)D) Wherein Ar is substituted aryl and one of the substituents is amino, to give compounds of the formula (II), (V), (XXII)A)，(XXIIC) Or (XXII)D) Wherein the substituent is NHCOR4 or NHSO2R4 residue. Preferably, the transformation is carried out under the same conditions as reported for step i').
According to the conversions described in 3), in various ways, the formulae (II), (V), (XXII) are reacted by reaction with the appropriate aldehyde or ketone, according to the customary methods for carrying out reductive alkylationA)，(XXIIC) Or (XXII)D) Wherein Ar is a substituted aryl group and one of the substituents is an amino group. Preferably, the reaction is carried out in a suitable solvent such as methanol, N-dimethylformamide, dichloromethane, tetrahydrofuran or mixtures thereof, in the presence of a suitable reducing agent such as sodium borohydride, tetraalkylammonium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, triacetoxytetramethylammonium borohydride, and an acidic catalyst such as acetic acid or trifluoroacetic acid, at a reaction temperature of from about 0 ℃ to reflux temperature, and for a reaction time varying from about 1 hour to about 96 hours.
According to the transformations described in 4), compounds of formula (I) wherein Ar is a substituted aryl group and one of the substituents is a nitro group, are reduced in various ways to compounds of formula (I) wherein the substituent is an amino group, according to conventional methods known in the literature. Preferably, the transformation is carried out under the same conditions as reported for transformation 1).
Acylating a compound of formula (I) wherein Ar is substituted aryl and one of the substituents is amino by reaction with an acetylating agent of formula (XXVII) or (XXVIII) wherein one of the substituents is NHCOR4 or NHSO according to the transformations described in 5) in various ways according to conventional methods well known in the literature to give compounds of formula (I)2R4 residue. Preferably, the transformation is carried out under the same conditions as reported for transformation 2).
According to the transformations described in 6), in various ways, the compounds of formula (I) in which Ar is a substituted aryl group and one of the substituents is an amino group are reductively aminated by reaction with an appropriate aldehyde or ketone according to conventional methods for carrying out reductive alkylation. Preferably, the transformation is carried out under the same conditions as reported for transformation 3).
It is known to the person skilled in the art that when compounds of formula (IV) or (XXVII) carry functional groups which may interfere with the acylation reaction, such groups must be protected before the reaction is carried out. In particular, when a compound of formula (IV) OR formula (XXVII) is substituted by a compound of formula NR5R6, OR7, SR7, R8R9N-C1-C6Alkyl or R8O-C1-C6When substituted by a residue of an alkyl group, wherein R7 or at least one of R5 and R6 or at least one of R8 and R9 represents hydrogen, such groups may be protected as is well known in the art. It is also known to those skilled in the art that such protecting groups can be removed immediately after the reaction or at a later stage in the synthesis process.
The formula (I), (XXII) can be reacted in various ways according to conventional methods for deprotecting amino groupsA)，(XXIICOr (XXII)D) Wherein Ar is a substituted aryl group and one of the substituents is a protected amino group. Depending on the amino protecting group, this reaction can be carried out in different ways. In one aspect, such reactions may be carried out by treatment with a mineral acid such as hydrochloric acid, sulfuric acid or perchloric acid or an organic acid such as trifluoroacetic acid or methanesulfonic acid in a suitable solvent such as water, methanol, ethanol, 1, 4-bisAlkane, tetrahydrofuran, diethyl ether, diisopropyl ether, acetonitrile, N, N-dimethylformamide, dichloromethane or a mixture thereof at a temperature of-10 ℃ to 80 ℃ for 30 minutes to 48 hours. In another aspect, this isThe analogous reaction can be carried out by treatment with an inorganic base such as lithium or sodium or potassium hydroxide, or sodium or potassium or cesium carbonate, or with anhydrous or hydrazine hydrate in a suitable solvent such as water, methanol, ethanol, 1, 4-diisopropylethylamine, or with an organic base such as triethylamine or N, N-diisopropylethylamineAlkane, tetrahydrofuran, diethyl ether, diisopropyl ether, acetonitrile, N, N-dimethylformamide, dichloromethane or a mixture thereof at a temperature of-10 ℃ to 80 ℃ for 30 minutes to 72 hours.
For example, Smith, Michael-March's Advanced Organic Chemistry: substituted indazole derivatives were prepared by standard methods in organic synthesis reported in interactions mechanisms and structures-5 th edition, Michael b. smith and Jerry March, john wiley & Sons inc, New York (NY), 2001. It is known to those skilled in the art that the conversion of one chemical functionality to another may require that one or more reactive centers on the compound containing this functionality be protected to avoid unwanted side reactions. May be as follows, for example: the standard methods described in Green, Theodora W. and Wuts, Peter G.M. -Protective Groups in Organic Synthesis, Third Edition, John Wiley & Sons Inc., New York (NY), 1999 protect such reaction centers and deprotect at the end of subsequent synthetic transformations.
In the case where the compound of formula (I) contains one or more asymmetric centers, the compound may be separated into individual isomers by methods well known to those skilled in the art. Such methods include standard chromatographic techniques, including chromatography or crystallization using a chiral stationary phase. For example, in Jacques, Jean; collet, Andre; general methods for the isolation of compounds containing one or more asymmetric centers are reported in Wilen, SamuelH., -Enantiomers, Racemates, and solutions, John Wiley & Sons Inc., New York (NY), 1981.
The compounds of formula (I) may also be converted into pharmaceutically acceptable salts according to standard procedures well known to those skilled in the art. Alternatively, the compounds of formula (I) obtained as salts may also be converted into the free base or free acid according to standard methods well known to those skilled in the art.
The starting materials for the process of the invention, i.e.the compounds of the formulae (XII), (XIII), (XV), (XVI), (XVIII), (XXIII) and (XXI), are commercially available or can be prepared by using well-known methods.
For example, the compounds of formula (XIII) are readily obtained according to conventional methods for grignard reagent reactions that are widely known to those skilled in the art, as reported in the following schemes:
RZ+Mg→RMgZ
(XIII)
for example, compounds of formula (XV) are readily prepared from the corresponding halogen derivatives as reported in the following schemes (see, e.g., WANG, X. -J., et al; Org Lett2006, 8(2), 305-307):
for example, the compound of formula (XVI) can be easily obtained by purifying the corresponding alcohol derivative by an operation according to a conventional synthetic method.
For example, the compounds of formula (XXI) are readily obtained by oxidation of the corresponding alcohol derivatives by procedures according to conventional synthetic methods.
It is another object of the present invention to provide formula (III)A')，(IIIB')，(IIIC') Or (III)D') The intermediate (2) of (a):
wherein R is optionally substituted C3-C6Cycloalkyl, aryl or heteroaryl, and
r1, R2, R3, R' and R ″ are as defined above, with the proviso that the following compounds are excluded:
-6- (3-amino-1H-indazol-5-ylmethyl) -3-isopropyl-1- (2,4, 6-trichloro-phenyl) -1, 7-dihydro-pyrazolo [3,4-d ] pyrimidin-4-one, and
-1- [ (3-amino-1H-indazol-5-yl) methyl ] -3- ({1- [2- (dimethylamino) ethyl ] -1H-benzimidazol-2-yl } methyl) -1, 3-dihydro-2H-benzimidazol-2-one.
It is another object of the present invention to provide compounds of formula (XII)A)，(XXIIC) Or (XXII)D) The intermediate (2) of (a):
wherein Ar, R, R1, R2, R3, R ', R ' ' and PG are as defined above.
It is another object of the present invention to provide a compound of formula (XXVII):
wherein Ar, R, R1, R2 and R3 are as defined above and PG2Is ethoxycarbonyl or 2-methoxyethylcarbonyl.
The present invention further provides a process for the preparation of a compound of formula (XXVII) as defined above, characterized in that it comprises:
v) protecting the compound of formula (I) as defined above to give a compound of formula (XXVII):
wherein R, R1, R2, R3 and PG2As defined above.
The protection of the compound of formula (I) to the compound of formula (XXVII) according to step v) can be carried out in various ways and experimental conditions. Preferably, the reaction is carried out by treatment with a base such as lithium diisopropylamide, sodium hydride or lithium bis (trimethylsilyl) amide, sodium bis (trimethylsilyl) amide or potassium bis (trimethylsilyl) amide in a suitable solvent such as toluene, tetrahydrofuran, 1, 4-bisAlkane, diethyl ether, N, N-dimethylformamide and dimethoxyethane, the reaction temperature is between-78 ℃ and room temperature, and the reaction time is between about 10 minutes and about 96 hours. The electrophile is typically a chloroformate derivative, for example, ethyl chloroformate or 2-methoxyethyl chloroformate.
Pharmacology of
The abbreviations and abbreviations used herein have the following meanings:
ci Curie
DMSO dimethyl sulfoxide
ID identity
KDa kilodalton
microCi micro Curie
mg of
microgram of microrog
mL of
micro L micro liter
M mol
mM millimole
Micromolar at μ M
nM nanomolar
Test of
The compounds of the invention were tested in biochemical assays, as described below.
Preparation of ALK cytoplasmic region for biochemical experiments
Cloning and expression
The ALK cytoplasmic region corresponding to residues 1060-1620 (the number of amino acid residues referenced to GenBank accession NP 004295.2) was PCR amplified from a human testis cDNA library.
Using a forward oligonucleotide: 5 'GGGGACAAGTTTGTACAAAAAAGCAGGCTTACTGGAAGTTCTGTTCCAGGGGCCCCGCCGGAAGCACCAGGAGCTG-3'
And a reverse oligonucleotide: 5'GGGGACCACTTTGTACAAGAAAGCTGGGTTTCAGGGCCCAGGCTGGTTCATGCTATT-3' for amplification.
For cloning purposes, the oligonucleotides included attB sites to obtain attB-flanked PCR products suitable for cloning using Gateway technology (Invitrogen). Furthermore, for purification purposes, the forward primer included a PreScission cleavage site (Amersham Biosciences). The resulting PCR product was cloned into Gateway-modified baculovirus expression vector pVL1393 (Invitrogen). For expression and purification purposes, a GST tag is added to the N-terminus of the cytoplasmic region of ALK. Cloning was performed according to the procedure described in the Gateway manual (Invitrogen).
By using BaculogoldTMThe baculovirus was generated by co-transfecting Sf9 insect cells with the expression vector and viral DNA using a transfection kit (Pharmingen).
After 5 days the virus supernatant was recovered and subjected to three rounds of amplification to increase virus titer.
30mL of viral supernatant per billion cells at 1X10 by shaking at 27 deg.C6cells/mLInfected Sf21 insect cells to produce recombinant proteins. After 48 hours of infection, the cells were recovered, pelleted and frozen at-80 ℃.
Protein purification
Cells were resuspended in lysis buffer (Tris-HCl50mM pH8, NaCl150mM, CHAPS0.2%, DTT20mM, Glycerol 20%, "complete" protease inhibitor cocktail (Roche Diagnostics), Na3VO41mM, and lysis was performed by squeezing the liquid out with a Gaulin homogenizer (Niro Soavi Italy). The lysate was clarified by centrifugation at 20000g for 30 minutes and loaded on a glutathione sepharose 4B (Amersham biosciences) column.
After thorough washing, the recombinant protein was eluted with 100mM Tris-HCl pH8, 10% glycerol, containing 10mM glutathione.
Loading of affinity purified GST-ALK on Heparin SepharoseTMFF (Amersham biosciences) column and eluted with 50mM NaCl, 25mM TRIS pH7.5, 2mM DTT, 20% glycerol.
Eluted fractions were combined and dialyzed against 150mM NaCl, 50mM Tris-HCl pH7.4, 2mM DTT, 20% glycerol.
The purified protein was stored at-80 ℃ prior to use in biochemical assays.
Biochemical assay for inhibitors of ALK kinase activity
The ALK enzyme requires preactivation to linearize the reaction kinetics.
i. Kinase Buffer (KB) for ALK
The kinase buffer consisted of 50mM HEPES pH7.5, containing 1mM MnCl2，5mM MgCl2，1mM DTT，3μMNa3VO4And 0.2mg/mL BSA. 3X KB is a buffer with the same composition and pH as KB, but the concentration of each component is three times greater.
Test conditions
The kinase assay was performed at 8. mu.M ATP, 1nM33P-. gamma. -ATP and 2. mu.M MBP were used at a final enzyme concentration of 20 nM. MPB was purchased from Sigma-Aldrich, St.Louis, MO, USA.
Cell-based assays for inhibitors of ALK kinase activity
Western blot analysis of phosphorylation of ALK and STAT3 in Karpas-299, SR-786 and SUP-M2 anaplastic large cell lymphoma cell lines
Karpas-299, SR-786 and SUP-M2 cells (DSMZ, Braunschwiegh, Germany) were cultured at 5X105cells/mL in RPMI-1640 medium +2mM glutamine +10% -15% FCS (EuroClone, Italy) seeded in 6-well tissue culture plates and 5% CO at 37 ℃2And incubated overnight at 100% relative humidity. After incubation, the cells were treated with the desired concentration of compound for 2 hours at 37 ℃. Cells were harvested by centrifugation at 248x g for 5 min, washed with cold PBS, centrifuged again at 248x g for 5 min, and then centrifuged at 100mM Tris-HCl pH7.4, 2% SDS, 1mM Na3VO4Protease inhibitor cocktail [ Sigma-Aldrich product # P8340]Phosphatase inhibitor cocktail [ Sigma-Aldrich products # P2850+ # P5726]) And (4) cracking. After a brief sonication, the cell lysate was clarified by centrifugation at 10,000Xg for 20 minutes at room temperature, and 20. mu.g/channel of clarified lysate protein was run on a NuPAGE gel (NuPAGE4-12% 10-channel Bis-Tris gel, Invitrogen) with MOPS running buffer, and then transferred to a Hybond-ECL nitrocellulose filter (Amersham Biosciences, Little Chalfount, Buckinghamshire, UK) using a MiniProTEAN II chamber (Bio-Rad Laboratories, Hercules, Calif., USA). Filters with transferred proteins were placed in blocking buffer (TBS +5% skimmed milk powder [ #1706404 Bio-rad, Hercules, CA, USA)]+0.1% Tween20) and introduction of probes overnight at 4 ℃ in TBS +5% BSA +0.1% Tween20 containing 1/500 anti-phospho-ALK Tyr1604 antibodies (product #3341Cell Signaling Technology, Beverly, MA, USA) for detection of phosphorylated ALK, or 1/500 mouse anti-ALK antibodies (product #35-4300, zymed laboratory)es, South San Francisco, CA, USA) was probed in TBS +5% BSA +0.1% Tween20 overnight for total ALK detection, or in TBS +5% BSA +0.1% Tween20 containing 1/500 mouse anti-phospho STAT3Tyr705 antibody (product #612357, bdtransmission Laboratories, Canada) overnight for phosphorylated STAT3, or in TBS +5% BSA +0.1% Tween20 containing 1/1000 mouse anti-STAT 3 antibody (product #610190BDTransduction Laboratories, Canada) overnight for total STAT 3.
In all cases, the filters were then washed with several changes of TBS +0.1% Tween20 for 20 minutes and incubated in TBS +5% skim milk powder +0.1% Tween20 containing 1/10000 dilutions of horseradish peroxidase conjugated anti-rabbit or mouse IgG (Amersham, product # NA934) before being washed again and developed using an ECL chemiluminescence system (Amersham) according to the manufacturer's recommendations. Unless otherwise indicated, reagents used were from Sigma-Aldrich, St.Louis, MO, USA.
In vitro cell proliferation assay for inhibitors of ALK kinase activity
Human ALCL cell lines Karpas-299, SR-786 and SUP-M2 at 1X105cells/mL were seeded in RPMI-1640 medium +2mM glutamine +10% -15% FCS (EuroClone, Italy), (100. mu.L/well) in 96-well plates (PerkinElmer, Wellesley, MA, USA) and maintained at 37 ℃ in 5% CO2100% relative humidity. The following day, plates were treated in duplicate with the appropriate dilutions of compounds starting from 10mM stock in DMSO (final DMSO concentration 0.1%). Eight untreated control wells were included in each plate. After 72 hours of treatment, 50 μ L of CellTiter-Glo Assay reagent (Promega, Madison, WI, USA) was added to each well, and the luminescence signal was measured after stirring using an Envision Detector (PerkinElmer Wellesley, MA, USA).
LSW/data analysis was performed using Microsoft Excel sigmoidal curve fitting, calculating IC50The value is obtained.
Preparation of IGF-1R for Biochemical assays
Cloning and expression
Human cDNA was used as a template for amplification by Polymerase Chain Reaction (PCR) of the predicted cytoplasmic portion of IGF-1R (amino acid residue 960-1367 of the precursor Protein; see NCBI Entrez Protein Access # P08069) including the entire kinase domain. PCR was performed using the forward primer sequence 5'-CTCGGATCCAGAAAGAGAAATAACAGCAGGCTG-3' and the reverse primer sequence 5'-CTCGGATCCTCAGCAGGTCGAAGACTGGGGCAGCGG-3'. To facilitate the subsequent cloning step, both primers included a BamHI restriction endonuclease site sequence. This PCR product was cloned in the shell frame using BamHI cohesive ends into the transfer vector of the baculovirus expression system pVL1392(Pharmingen) previously modified by insertion into the pVL1392 multiple cloning site of the sequence encoding the glutathione S-transferase (GST) fusion protein, the PreScission protease cleavage site and a partial MCS cassette from the pGex-6P plasmid (Amersham BioSciences). The above IGF-1R PCR product was inserted into the pGex-6P-derived BamHI site of the modified pVL1392 vector, generating an open reading frame corresponding to pGEX-6P GST protein and PreScission peptide fused to the cytoplasmic domain of human IGF-1R. To obtain the fusion protein, Sf21 insect cells (Invitrogen) were co-transfected with 2. mu.g of purified plasmid and 1. mu.g of viral DNA (BaculoGoldTM Transfection Kit, Pharmingen) as described in Baculovir instruments manual (Pharmingen). First amplification of the Virus 600. mu.l of the co-transfected virus were used, 6X10 in monolayer culture in 12mL medium (TNM-FH Grace medium-Pharmingen)6Sf 21. After 3 days, the medium was collected, centrifuged and transferred to a sterile tube. The same method was used with 3X10 diluted in 40mL of medium72mL on cells to prepare a second expansion. For the third amplification of the virus, for every 3 × 10 diluted in 40mL of medium7Cells, 1mL of supernatant from the second round was used.
After using 14mL of virus/1X 109Protein expression in insect cell (MOI =1.5) infected H5 insect cells, shaken at 27 deg.CThe reaction was carried out under agitation for 65 hours. Cells were harvested by centrifugation at 1200Xg for 10 min.
Protein purification
Cells were resuspended in Phosphate Buffered Saline (PBS), 20mM Dithiothreitol (DTT), 0.2% CHAPS, 20% glycerol, 1mM OVA, "complete" protease inhibitor cocktail (1 tablet/50 mL buffer; Roche Diagnostics, Milan, Italy) and lysed by squeezing the liquid out with a Gaulin homogenizer (Niro Soavi, Italy). The lysate was centrifuged at 14000Xg for 45 minutes and the supernatant was loaded onto a column containing 10mL glutathione Sepharose (Amersham Biosciences). The column was first washed with 5 column volumes of PBS buffer, then 5 column volumes of 100mM Tris pH8.0, 20% glycerol, and finally eluted with 100mM Tris pH8.0, 20% glycerol containing 10mM glutathione. Fractions of 10mL were collected and protein rich fractions were pooled. Typically from 1x10920mg of fusion protein was recovered from the cells, which was typically judged by SDS-PAGE and subsequent Coomassie staining>85% purity. The purified protein was stored at-80 ℃ prior to use in biochemical assays.
Biochemical assays for inhibitors of IGF-1R kinase Activity
The putative kinase inhibitor inhibitory activity as well as the potency of the selected compound was determined using the transphosphorylation assay.
In a trace amount33P-gamma-ATP (gamma phosphate-labeled, Redivue)TMATP coding for AH9968, 1000-3000Ci/mmole, Amersham Biosciences Piscataway, NJ, USA), optimal cofactors and test compounds were incubated with the specific substrate and kinase in the presence of appropriate buffer conditions.
At the end of the phosphorylation reaction, more than 98% of the cold and radioactive ATP was captured by excess Dowex ion exchange resin. The resin is allowed to settle to the bottom of the reaction well by gravity. The supernatant containing the substrate peptide was then withdrawn and transferred to a counting plate and the radioactivity (corresponding to the phosphate incorporated into the peptide) was assessed by beta-counting.
Reagent/assay conditions
Preparation of Dowex resin
500g of wet resin (SIGMA, DOWEX resin 1X8200-400 mesh, 2.5Kg, conventionally prepared) was weighed and diluted into 2L of 150mM sodium formate, pH 3.00.
The resin was allowed to settle for several hours and the supernatant was discarded. This washing operation was repeated three times over two days. Finally, the resin was precipitated, the supernatant discarded and two volumes (relative to the resin volume) of 150mM sodium formate buffer were added. The final pH was about 3.0. The washed resin was kept at 4 ℃ until use and it was stable for more than one week.
Kinase Buffer (KB)
The kinase buffer consisted of 50mM HEPES pH7.9, containing 3mM MgCl2，1mMDTT，3μM Na3VO4And 0.2mg/mL BSA. 3X KB is a buffer with the same composition and pH as KB, but the concentration of each component is three times that of KB.
Enzyme preactivation and preparation of 3X enzyme mixture.
IGF-1R was pre-phosphorylated prior to initiation of the kinase inhibition assay to linearize the reaction kinetics. To this end, the required total amount of enzyme was prepared in KB containing 100. mu.M ATP at an enzyme concentration of 360nM and the preparation was incubated for 30min at 28 ℃. The 3X enzyme mixture was obtained by diluting the pre-activated enzyme 20-fold in 3X KB.
Test conditions
The kinase assay was performed with a final enzyme concentration of 6nM, at 6. mu.M ATP, 1nM33P- γ -ATP and 10 μ M substrate, having the sequence: KKKSPGEYVNIEFGGGGGK-in the presence of a biotinylated peptide at the carboxy terminus of biotin. The peptide is represented by>The 95% Peptide purity was obtained in portions from American Peptide Company, Inc (Sunnyvale, CA, USA).
Automated Dowex test
The test reactions were carried out in a total final volume of 21 μ Ι _ comprising:
a)7 μ L/well of 3 Xenzyme mix (18 nM pre-activated enzyme in 3 Xkinase buffer),
b)7 μ L/well of 3 Xsubstrate/ATP mix (in double distilled Water (ddH)2O) 30. mu.M substrate, 18. mu.M ATP, 3nM33P-γ-ATP)，
c) Dilution to ddH 7. mu.L/well2Test compounds at 3X in O-3% DMSO.
Compound dilution and test protocol is reported below
i. Dilution of the Compound
Test compounds in 100% DMSO 10mM stock solution distribution in 96 hole 12x8 format microtiter plate.
For% inhibition studies, 1mM, 100. mu.M and 10. mu.M dilution plates were prepared in 100% DMSO, then in ddH2O, 3% DMSO diluted to 3X final desired concentration (30, 3 and 0.3 μ M). Multimek96(Beckman Coulter, Inc.4300N.harbor Boulevard, P.O.Box3100Fullerton, CA92834-3100USA) was used to pipette compounds into the assay plates.
For the IC50 assay, starting solutions of 30. mu.M compound in 3% DMSO were obtained from 1mM/100% DMSO stock. These 30. mu.M starting solutions were used in ddH2Additional 9 serial 1/3 dilutions were made in O, 3% DMSO to give a 10-point dilution curve of 3X final assay concentration. Serial dilutions were performed in 96-well plates using a Biomek2000(Beckman Coulter) system. Dilution curves were prepared for 7 compounds per plate, and each plate also included a 10-point dilution curve for the astrosporin and several negative and positive control wells.
Test protocol
mu.L of each test compound (or control) in ddH2Dilution in O, 3% DMSOThe solution was pipetted into each well of a 384 well V-bottom assay plate and then transferred to a PlateTrak12 automated workstation (Perkin Elmer, 45William Street Wellesley, MA02481-4078, USA) equipped with a 384-ended pipetting head for initiating the assay, and a 96-ended head for dispensing the resin, provided with reservoirs containing sufficient 3X enzyme mix and 3X ATP mix (3X) to complete the assay process.
At the start of the experiment, the liquid handling system aspirates 7 μ L of ATP mix, introduces an air gap (5 μ L) into the tip, and then aspirates 7 μ L of 3 Xenzyme mix. To start the reaction, the end contents were dispensed into test wells that already contained 7 μ Ι _ of test compound (at 3X the desired final concentration), followed by 3 mixing cycles to reestablish the desired final concentration of all reaction components.
The plates were incubated at room temperature for 60 minutes, then the reaction was stopped by pipetting 70 μ L of Dowex resin suspension into the reaction mixture, followed by three mixing cycles. After terminating the reaction, the plate was left for 1 hour to maximize capture of ATP. At this point, 20. mu.L of supernatant was transferred from each well to 384-Optiplates (Perkin Elmer) wells containing 70. mu.L/well of Microscint40(Perkin Elmer); after orbital oscillation for 5 minutes, plates were read on a Perkin-Elmer Top Count radioactivity counter.
Analysis of data
Data were analyzed using a specialized version of the "Assay Explorer" software package (Elsevier MDL, San Leandro, CA 94577). For individual compound concentrations, the inhibitory activity is typically expressed as% inhibition obtained in the presence of the compound compared to the total activity of the enzyme obtained in the absence of the inhibitor.
The compounds exhibiting the desired inhibitory effect are further analyzed for passage of IC50Calculated to study the efficacy of the inhibitor. In this case, inhibition data obtained using serial dilutions of the inhibitor was fitted by non-linear regression using the following equation：
Wherein vbIs the baseline velocity, v is the observed reaction velocity, voIs the speed in the absence of inhibitor, and [ I]Is the inhibitor concentration.
Cell-based assays for inhibitors of IGF-1R kinase activity
Western blot analysis of receptor phosphorylation after stimulation with IGF-1 in MCF-7 human breast cancer cells
MCF-7 cells (ATCC # HTB-22) at 2X105Cells/well E-MEM Medium (MEM + Earle's BSS +2mM glutamine +0.1mM non-essential amino acids) +10% FCS seeded in 12-well tissue culture plates and 5% CO at 37 ℃%2And incubated overnight at 100% relative humidity. Cells were then starved by replacing E-MEM +10% FCS with E-MEM +0.1% BSA and cultured overnight. After incubation, wells were treated with the desired concentration of compound for 1 hour at 37 ℃ and then stimulated with 10nM recombinant human IGF-1(Invitrogen, Carlsbad, Calif., USA) for 10 minutes at 37 ℃. The cells were then washed with PBS and lysed in 100. mu.L/well of cell lysis buffer (M-PER mammalian protein extraction reagent [ product #78501, Pierce, Rockford, IL, USA)]+10mM EDTA + protease inhibitor cocktail [ Sigma-Aldrich product # P8340]+ phosphatase inhibitor cocktail [ Sigma-Aldrich products # P2850+ # P5726]) Dissolving in water. Cell lysates were clarified by centrifugation at 10,000Xg for 5 minutes and 10. mu.g/channel of clarified lysate proteins were run on a NuPAGE gel (NuPAGE4-12% 10-channel Bis-Tris gel, Invitrogen) with MOPS running buffer and then transferred to a Hybond-ECL nitrocellulose filter (Amersham Biosciences, Little Chalfount, Buckingshihire, UK) using a Mini PROTEAN II chamber (Bio-Rad Laboratories, Hercules, CA, USA). The filters with transferred proteins were incubated for 1 hour in blocking buffer (TBS +5% BSA +0.15% Tween20) and in a medium containing 1/1000 rabbit anti-phosphateThe probes were introduced in the same buffer of IGF-1R Tyr1131/InsR Tyr1146 antibody (product #3021, Cell Signaling technology, Beverly, MA, USA) for 2 hours for detection of phosphorylated IGF-1R, or in the same buffer containing 1/1000 dilutions of rabbit IGF-Ir β (H-60) antibody (product # sc-9038, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) for 2 hours for detection of total IGF-1R β chains in both cases, the filters were then washed with TBS +0.15% Tween20 for several changes for 30 minutes and incubated in a wash buffer containing 1/5000 degrees of horseradish peroxidase conjugated anti-rabbit IgG (Amersham, product # NA934) for 1 hour, then washed again and developed using the chemiluminescence system (Amersham) according to the manufacturer, unless otherwise stated, Sigma from Sigma, USA.
Growth factor-induced phosphorylation of S6 ribosomal protein in primary human fibroblasts
The phosphorylation of S6 ribosomal protein in response to growth factor stimulation of Normal Human Dermal Fibroblasts (NHDF) was used to assess the efficacy of compounds in inhibiting IGF-1 induced signaling in cells, as well as the selectivity for EGF and PDGF stimulation. NHDF cells from Promocell (Heidelberg, Germany) were maintained at 37 ℃ in the presence of 5% CO2In complete Fibroblast Growth Medium (PromoCell). For the assay, NHDF were seeded at 5000 cells/well in 384-well tissue culture plates (clear flat bottom black plates; Matrix Technologies Inc., Hudson, NH, USA) in serum free medium containing 0.1% Bovine Serum Albumin (BSA) and cultured for 5 days. Starved cells were treated with the required dose of compound for 1 hour, then restimulated with 10nM IGF-1(Invitrogen Corp., CA, USA), 10nM EGF (Gibco BRL, USA) or 1nM PDGF-B/B (Roche Diagnostics GmbH, Germany) for 2 hours. Cells were then fixed in PBS/3.7% paraformaldehyde for 20 minutes at room temperature, washed 2 times with PBS, and permeabilized with PBS/0.3% Triton X-100 for 15 minutes. Wells were then washed with PBS/1% skim milk powder (Bio-Rad Laboratories, Hercules, CA,USA) were saturated for 1 hour, and then the probes were introduced with 1/200 dilutions of anti-phospho-S6 (Ser235/236) antibody (CellSignaling Technology, Beverly, MA, USA, cat. #2211) in PBS/1% milk/0.3% Tween20 at 37 ℃. The wells were then washed twice with PBS and at 37 ℃ with PBS/1% milk/0.3% Tween20+ 1. mu.g/mL DAPI (4, 6-diamidino-2-phenylindole) +1/500 goat anti-rabbit Cy5TMBound secondary antibody (Amersham Biosciences, Little Chalfount, Buckinghamshire, UK) was incubated for 1 hour. Wells were then washed 2 times with PBS and 40 μ Ι _ of PBS was dispensed into each well for immunofluorescence analysis. Obtained automatically at DAPI and Cy5TMFluorescence image in channel using CellomicrsrrrayScanTMIV instrument (Cellomics, Pittsburgh, USA) storage and analysis; quantification of phospho-S6 using Cellomics cytoxicity Algorithm with 10 fields/well per cell (Cy 5)TMSignal parameters: "MeanLyso Mass-pH") and finally expressed as mean population values. Unless otherwise indicated, reagents were from Sigma-Aldrich, St.Louis, MO, USA.
Biochemical assay for inhibitors of Aurora-2 kinase activity
In vitro kinase inhibition assays were performed in the same manner as described for IGF-1R. However, it differs from IGF-1R in that the Aurora-2 enzyme does not require prior activation.
i. Kinase Buffer (KB) for Aurora-2
The kinase buffer consists of 50mM HEPES, pH7.0, 10mM MgCl2，1mM DTT，3μM Na3VO4And 0.2mg/mL BSA.
Test conditions for Aurora-2 (final concentration)
Kinase assay used an enzyme concentration of 2.5nM, 10. mu.M ATP, 1nM33P-gamma-ATP and 8. mu.M substrate, consisting of 4 LRRWSSG repeats
Cell-based assays for inhibitors of Aurora-2 kinase activity
In vitro cell proliferation assay for inhibitors of Aurora-2 kinase activity
Human colon cancer cell line HCT-116 at 5000 cells/cm2Inoculated in 24-well plates (Costar), using F12 medium (Gibco) supplemented with 10% FCS (EuroClone, Italy)2mM L-glutamine and 1% penicillin/streptomycin, and maintained at 37 ℃ with 5% CO2And 96% relative humidity. The following day, plates were treated with 5mL of compound in duplicate at appropriate dilutions starting from a 10mM stock in DMSO. Two untreated control wells were included in each plate. After 72 hours of treatment, the medium was aspirated and 0.5mL of 0.05% (w/v) Trypsin, 0.02% (w/v) EDTA (Gibco) was used to detach the cells from each well. The samples were diluted with 9.5mL of isoton (Coulter) and counted using a Multisizer3 cell counter (Beckman Coulter). Data were evaluated as a percentage of control wells:
CTR% = (treated-blank)/(control-blank).
LSW/data analysis was performed using Microsoft Excel sigmoidal curve fitting, calculating IC50The value is obtained.
From the above experiments it was concluded that the compounds of formula (I) according to the invention lead to a significant protein kinase inhibitory activity, typically with an IC below 10. mu.M50。
For example, see Table 1 below, where experimental data (IC) are reported for certain representative compounds of the invention assayed as ALK, IGF-1R and Aurora-2 kinase inhibitors in biochemical assays50μM)。
Table 1.
As can be seen from all the above results, the novel compounds of formula (I) according to the invention show particular advantages in the treatment of diseases resulting from a dysregulated protein kinase activity, such as cancer.
The compounds of the invention may be administered as the sole active agent or, alternatively, in combination with known anti-cancer therapies such as radiation therapy or chemotherapeutic regimens, for example, with anti-hormonal agents (e.g., anti-estrogenic agents, anti-androgens, and aromatase inhibitors), topoisomerase I inhibitors, topoisomerase II inhibitors, microtubule-targeting agents, platinum-based agents, alkylating agents, DNA damaging or intercalating agents, antineoplastic agents, antimetabolite agents, other kinase inhibitors, other anti-angiogenic factors, inhibitors of kinesin, therapeutic monoclonal antibodies, inhibitors of mTOR, histone deacetylase inhibitors, farnesyl transferase inhibitors, and inhibitors of hypoxia response.
If formulated as a fixed dose, such combination products employ the compounds of the present invention within the dosage ranges described below and other pharmaceutically active agents within the approved dosage ranges.
When the combined preparation is not suitable for use, the compound of formula (I) may be used in combination with a known anticancer agent.
The compounds of formula (I) of the present invention, which are suitable for administration to mammals (e.g., to humans), can be administered by conventional routes, and the dosage level depends on the age, weight and condition of the patient and the route of administration.
For example, a suitable dose for oral administration of a compound of formula (I) may be from about 10 to about 500mg per dose, 1-5 times per day. The compounds of the invention may be administered in a variety of dosage forms, for example orally in the form of tablets, capsules, sugar-or film-coated tablets, liquid solutions or suspensions; rectally in the form of suppositories; parenteral administration is, for example, intramuscular injection, or by intravenous and/or intrathecal and/or intraspinal injection or infusion.
The invention also includes pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, which may be a carrier or diluent.
Pharmaceutical compositions comprising the compounds of the invention are generally prepared according to conventional methods and administered in suitable pharmaceutical forms.
For example, solid oral forms may comprise the active compound, together with diluents, e.g., lactose, glucose, disaccharides, sucrose, cellulose, corn starch or potato starch; lubricants, for example, silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glycol; binders, for example, starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disintegrants, for example, starch, alginic acid, alginates, or sodium starch glycolate; an effervescent mixture; a dye; a sweetener; wetting agents, such as lecithin, polysorbate, lauryl sulfate; and nontoxic and pharmacologically inactive substances commonly used in pharmaceutical preparations. These pharmaceutical preparations can be produced in a known manner, for example, by mixing, granulating, tabletting, sugar-coating, or film-coating methods.
Liquid dispersions for oral administration may be, for example, syrups, emulsions and suspensions.
For example, the syrup may comprise sucrose or sucrose together with glycerol and/or mannitol and sorbitol as a carrier.
Suspensions and emulsions may contain, as carrier, for example, natural gums, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol.
Suspensions or solutions for intramuscular injections may contain the active compound in association with a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and, if desired, a suitable amount of lidocaine hydrochloride.
Solutions for intravenous injection or infusion may contain sterile water as a carrier, or preferably they may be in the form of sterile, aqueous, isotonic saline solutions, or they may contain propylene glycol as a carrier.
Suppositories may contain the active compound together with a pharmaceutically acceptable carrier, for example, cocoa butter, polyethylene glycol, a polyoxyethylene sorbitan fatty acid ester surfactant or lecithin.
Experimental part
For any particular compound of formula (I), optionally in pharmaceutically acceptable salt form, of the invention, see experimental section and claims. For the following examples, the compounds of the invention are synthesized using the methods described herein or other methods well known in the art.
The phrases and abbreviations used herein have the following meanings:
in order to better illustrate the invention, without making any limitation thereto, the following examples are given.
The symbols used herein and the rules used in the methods, protocols and examples are consistent with those used in the existing scientific literature, e.g., the Journal of the American Chemical Society or Journal of Biological Chemistry.
Unless otherwise noted, all starting materials were obtained from commercial suppliers, had the best grades and were used without further purification. Anhydrous solvents, e.g. DMF, THF, CH2Cl2And toluene from Aldrich chemical company. All reactions involving air or moisture sensitive compounds were carried out in a nitrogen or argon atmosphere.
General purification and analytical methods
Flash chromatography was performed on silica gel (Merck grade9395, 60A). HPLC was performed on a Waters X Terra RP18(4.6X50mm, 3.5 μm) column using a Waters2790HPLC system equipped with a 996Waters PDA detector and a Micromass mod.zq single quadrupole mass spectrometer equipped with an Electrospray (ESI) ion source. Mobile phase a was ammonium acetate 5mM buffer (ph 5.5 adjusted with acetic acid-acetonitrile 95: 5) and mobile phase B was water-acetonitrile (5: 95). The gradient was from 10-90% B over 8 min, held at 90% B for 2 min. UV detection was performed at 220nm and 254 nm. The flow rate was 1 mL/min. The injection volume was 10. mu.L. A full scan was performed from a mass range of 100-. The capillary voltage was 2.5 KV; the source temperature was 120 ℃; the cone voltage is 10V. The retention time (HPLC, room temperature) is given in minutes, at 220nm or at 254 nm. The mass is expressed as the m/z ratio.
If desired, compounds were purified by preparative HPLC on a Waters Symmetry C18(19X50mm, 5 μm) column or a Waters X TerraraP 18(30X150mm, 5 μm) column using Waters preparative HPLC600 equipped with a 996Waters PDA detector and a Micromass mod.ZMD single quadrupole mass spectrometer using electrospray ionization, anodic mode. Use of: acid method, mobile phase A is water-0.01% trifluoroacetic acid, mobile phase B is acetonitrile; gradient from 10-90% B in 8 min, hold at 90% B for 2 min; the flow rate was 20 mL/min. Or, alkaline process, mobile phase A is water-0.1% NH3Mobile phase B is acetonitrile; gradient from 10-100% B in 8 min, hold at 100% B for 2 min; the flow rate was 20 mL/min.
1H-NMR spectroscopy was performed on Mercury VX400 operating at 400.45MHz equipped with a 5mm dual resonance probe [1H (15N-31P) ID _ PFGiarian ].
Example 1
Step a
5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -2-fluoro-benzonitrile [ (XI), R1= R2= R3= H, R =3, 5-difluorophenyl ]
To a stirred suspension of magnesium powder (2.6g, 109mmol) in anhydrous tetrahydrofuran (10mL) under an argon atmosphere was slowly added a solution of 1-bromo-3, 5-difluoro-benzene (21g, 109mmol) in dry tetrahydrofuran (90 mL). The reaction mixture was stirred and heated at 90 ℃ until all magnesium was consumed (1 hour). Thereafter, the reaction was cooled at-10 ℃ and a solution of 2-fluoro-5-formyl-benzonitrile (13.5g, 90.6mmol) in 100mL of anhydrous tetrahydrofuran was added over the course of 30 min. After 1 hour, the reaction mixture was quenched by dropwise addition of 200mL of 20% ammonium chloride solution. Ethyl acetate was added, the layers were separated and the aqueous layer was extracted twice with ethyl acetate. The organic layer was collected, washed with brine, dried and evaporated. The crude product was reacted with isopropyl ether/hexane 1: 1(100mL) were triturated together, filtered and washed with the same mixture (50mL) to give 16g of the final product. The resulting organic phase was purified by silica gel chromatography (hexane/EtOAc 4: 1) to give 4.5g of the title compound (total 20.5g, 87% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：5.82(d，J=4.02Hz，1H)6.41(d，J=4.02Hz，1H)7.05-7.12(m，1H)7.12-7.18(m，2H)7.46-7.50(m，1H)7.80(td，J=5.76，2.62Hz，1H)7.97(dd，J=6.34，2.19Hz，1H)
The following compounds were obtained by operating in the same manner:
5- (phenyl-hydroxy-methyl) -2-fluoro-benzonitrile [ (XI), R1= R2= R3= H, R = phenyl ]
ESI(+)MS：m/z245(MNH4 +)。
Step b
5- (3, 5-difluoro-benzoyl) -2-fluoro-benzonitrile [ (X), R1= R2= R3= H, R =3, 5-difluorophenyl ]
A mixture of 5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -2-fluoro-benzonitrile (2.68g, 10.2mmol), 4-methylmorpholine N-oxide monohydrate (2.02g, 15mmol) and tetrapropylammonium perruthenate (35mg, 0.1mmol) in dry dichloromethane (50mL) was stirred at room temperature for 2 hours. The reaction mixture was evaporated and the residue was redissolved in ethyl acetate. The organic phase was washed with 10% sodium bisulfite and saturated ammonium chloride, dried and evaporated. The crude product was purified by silica gel chromatography (EtOAc/hexanes) to give 2.05g of the title compound (77% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.43-7.50(m，2H)7.61-7.68(m，1H)7.72(t，J=9.02Hz，1H)8.17(ddd，J=8.84，5.30，2.32Hz，1H)8.35(dd，J=6.22，2.20Hz，1H)
The following compounds were obtained by operating in the same manner:
5-benzoyl-2-fluoro-benzonitrile [ (X), R1= R2= R3= H, R = phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.59(t，J=7.81Hz，2H)7.72(m，2H)7.78(dd，J=8.30，1.46Hz，2H)8.13(ddd，J=8.79，5.37，2.20Hz，1H)8.28(dd，J=6.10，2.20Hz，1H)
Step c
(3-amino-1H-indazol-5-yl) - (3, 5-difluoro-phenyl) -methanone [ (IX), R1= R2= R3= H, R =3, 5-difluorophenyl ]
A mixture of 5- (3, 5-difluoro-benzoyl) -2-fluoro-benzonitrile (2.05g, 7.84mmol) and hydrazine hydrate (0.73mL, 15.7mmol) in dry tetrahydrofuran (100mL) was stirred at room temperature for 2 hours. The reaction mixture was treated with 37% hydrochloric acid (1.3mL, 15.7mmol) for 30min, then the volatiles were partially evaporated. The reaction mixture was then diluted with water (100mL) and NH was added3Water to reach a neutral pH. The resulting solid was filtered, washed well with water and dried under vacuum at 60 ℃. The title compound was obtained as a yellow solid (1.75g, 80% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：5.75(br.s.，2H)7.33-7.36(m，1H)7.36-7.40(m，2H)7.52-7.59(m，1H)7.75(dd，J=8.84，1.65Hz，1H)8.27(dd，J=1.59，0.73Hz，1H)11.95(br.s.，1H)
The following compounds were obtained by operating in the same manner:
(3-amino-1H-indazol-5-yl) - (3-ethoxy-phenyl) -methanone [ (IX), R1= R2= R3= H, R = 3-ethoxyphenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.36(t，J=7.01Hz，3H)4.11(q，J=6.95Hz，2H)7.20-7.24(m，2H)7.25-7.28(m，1H)7.41(dd，J=8.84，0.55Hz，1H)7.48(td，J=7.68，0.61Hz，1H)7.80(dd，J=8.78，1.59Hz，1H)8.34(d，J=0.85Hz，1H)12.24(br.s.，1H)
(3-amino-1H-indazol-5-yl) -phenyl-methanone [ (IX), R1= R2= R3= H, R = phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.40(dd，J=8.78，0.61Hz，1H)7.57(tt，J=7.68，1.59Hz，2H)7.66(tt，J=7.32，2.07Hz，1H)7.72(dt，J=6.83，1.34Hz，2H)7.78(dd，J=8.78，1.59Hz，1H)8.31(m，1H)12.15(br.s.，1H)
Step d
N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (VIII), R1= R2= R3= H, R =3, 5-difluorophenyl, PG1= trifluoroacetyl group]
A suspension of (3-amino-1H-indazol-5-yl) - (3, 5-difluoro-phenyl) -methanone (2.73g, 10mmol) in dry tetrahydrofuran (120mL) was treated with trifluoroacetic anhydride (4.2mL, 30mmol) and stirred at room temperature for 1 hour. The solution was evaporated, treated with methanol and further evaporated to dryness. The residue was redissolved in ethyl acetate and washed with aqueous bicarbonate solution. The organic phase was separated, dried and evaporated. The solid was triturated with a small amount of dichloromethane and filtered to give 3.25g (88% yield) of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.39-7.46(m，2H)7.56-7.64(m，1H)7.68(dd，J=8.84，0.67Hz，1H)7.86(dd，J=8.84，1.65Hz，1H)8.28-8.32(m，1H)12.16(s，1H)13.50(s，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3-ethoxy-benzoyl) -1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (VIII), R1= R2= R3= H, R = 3-ethoxyphenyl, PG1= trifluoroacetyl group]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34(t，J=6.95Hz，3H)4.10(q，J=6.95Hz，2H)7.19-7.25(m，2H)7.28(d，J=7.56Hz，1H)7.43-7.50(m，1H)7.67(d，J=8.90Hz，1H)7.85(dd，J=8.84，1.52Hz，1H)8.26(s，1H)12.14(s，1H)13.46(s，1H)
Step e
N- [5- (3, 5-difluoro-benzoyl) -1-trityl-1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (VII), R1= R2= R3= H, R =3, 5-difluorophenyl, PG = triphenylmethyl, PG1= trifluoroacetyl group]
N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl ] in dry dichloromethane (300mL) was treated with chlorotriphenylmethane (14.72g, 52.8mmol) and triethylamine (14.55mL, 103.5mmol)]-2,2, 2-trifluoro-acetamide (19.11g, 51.76 mmol). After stirring at room temperature for 2 days, with NH4The reaction was washed with Cl solution, dried and evaporated. The crude product was purified by silica gel chromatography (DCM/MeOH) to give 27.32g of the title compound (86% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：6.57(d，J=8.90Hz，1H)7.20(m，6H)7.29-7.40(m，11H)7.58(m，2H)8.22(d， J=1.10Hz，1H)12.27(s，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3-ethoxy-benzoyl) -1-trityl-1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (VII), R1= R2= R3= H, R = 3-ethoxyphenyl, PG = triphenylmethyl, PG1= trifluoroacetyl group]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33(t，J=6.95Hz，3H)4.10(q，J=6.95Hz，2H)6.56(d，J=9.02Hz，1H)7.17-7.39(m，18H)7.41-7.47(m，1H)7.54(dd，J=9.08，1.65Hz，1H)8.18(d，J=0.98Hz，1H)12.25(s，1H)
Step f
(3-amino-1-trityl-1H-indazol-5-yl) - (3, 5-difluoro-phenyl) -methanone [ (VI), R1= R2= R3= H, R =3, 5-difluorophenyl, PG = triphenylmethyl ]
N- [5- (3, 5-difluoro-benzoyl) -1-trityl-1H-indazol-3-yl ] -2,2, 2-trifluoro-acetamide (6.12g, 10mmol) was reacted at 100 ℃ in isopropanol/tetrahydrofuran 8: 2(100mL) and triethylamine (12.2mL) for 48 hours. The volatile material was partially evaporated, cooled and the resulting mixture filtered. The solid was washed with diethyl ether. After drying in vacuo at 70 ℃ the title compound was obtained as a white solid (5.1g, 99% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：5.98(br.s.，2H)6.35(d，J=8.90Hz，1H)7.20-7.37(m，17H)7.48(dd，J=9.08，1.77Hz，1H)7.50-7.57(m，1H)8.23(d，J=1.10Hz，1H)
The following compounds were obtained by operating in the same manner:
(3-amino-1-trityl-1H-indazol-5-yl) - (3-ethoxy-phenyl) -methanone [ (VI), R1= R2= R3= H, R = 3-ethoxyphenyl, PG = triphenylmethyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33(t，J=6.95Hz，3H)4.07(q，J=6.95Hz，2H)5.93(s，2H)6.36(d，J=9.02Hz，1H)7.12-7.34(m，18H)7.40-7.46(m，2H)8.22(d，J=1.10Hz，1H)
Step g
N- [5- (3, 5-difluoro-benzoyl) -1-trityl-1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (V), R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl, PG = triphenylmethyl ]
To a suspension of 4- (4-methyl-piperazin-1-yl) -2-nitro-benzoic acid hydrochloride (1.5g, 4.97mmol) in dry tetrahydrofuran (80mL) were added oxalyl chloride (1.4mL, 19.9mmol) and N, N-dimethylformamide (1-2 drops). The mixture was stirred at room temperature overnight and then evaporated to dryness. The resulting crude acid chloride was dissolved in toluene and evaporated again, then dissolved in dry tetrahydrofuran (180 mL). A solution of (3-amino-1-trityl-1H-indazol-5-yl) - (3, 5-difluoro-phenyl) -methanone (1.83g.3.55mmol) and N, N-diisopropylethylamine (2.5mL, 14.22mmol) in dry tetrahydrofuran (15mL) was added to the reaction mixture. The mixture was stirred at room temperature overnight and then at 75 ℃ for 2 hours. The volatiles were evaporated and the residue was dissolved in dichloromethane and washed with brine. The organic phase was dried over sodium sulfate and evaporated to dryness. The crude product was purified by silica gel chromatography (DCM/MeOH) to give 2.51g of the title compound as a yellow powder (92% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.22(s，3H)2.40-2.45(m，4H)3.26-3.36(m，4H)6.50(d，J=8.17Hz，1H)7.19-7.50(m，21H)7.56(dd，J=9.15，1.71Hz，1H)8.28-8.30(m，1H)11.22(br.s.，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3-ethoxy-benzoyl) -1-trityl-1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (V), R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl, PG = triphenylmethyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34(t，J=6.95Hz，3H)2.24(m，3H)2.45(m，4H)3.27(m，4H)4.08(q，J=6.95Hz，2H)6.51(d，J=8.17Hz，1H)7.20-7.46(m，22H)7.53(dd，J=9.15，1.71Hz，1H)8.30(m，1H)11.22(br.s.，1H)
Step h
N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (II), R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl ]
A mixture of N- [5- (3, 5-difluoro-benzoyl) -1-trityl-1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide (2.76g, 3.62mmol), trifluoroacetic acid (5.6mL) and dichloromethane (56mL) was stirred at room temperature for 2 hours. The volatiles were evaporated and the residue was dissolved in dichloromethane and washed with saturated sodium bicarbonate solution. The organic phase was evaporated to dryness. The residue was redissolved in ethyl acetate and washed twice with brine. The organic phase obtained is dried over sodium sulfate and evaporated to dryness. The crude product was purified by silica gel chromatography (DCM/MeOH), and the compound thus obtained was triturated from ether to yield 1.47g of the title compound (78% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.25(br.s.，3H)2.47(br.s.，4H)3.29-3.38(m，4H)7.26(dd，J=8.84，2.50Hz，1H)7.37-7.43(m，2H)7.45(d，J=2.44Hz，1H)7.51-7.59(m，1H)7.63(dd，J=8.84，0.55Hz，1H)7.66(br.s.，1H)7.86(dd，J=8.84，1.65Hz，1H)8.36(s，1H)11.13(s，1H)13.21(s，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3-ethoxy-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (II), R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34(t，J=6.95Hz，3H)2.26-2.34(m，3H)2.46-2.59(m，4H)3.28-3.35(m，4H)4.10(q，J=6.95Hz，2H)7.18-7.21(m，1H)7.24-7.26(m，1H)7.27(dd，J=9.33，1.89Hz，1H)7.29-7.32(m，1H)7.45(t，J=7.87Hz，1H)7.46(d，J=2.32Hz，1H)7.62(d，J=9.02Hz，1H)7.66(d，J=9.88Hz，1H)7.84(dd，J=8.78，1.59Hz，1H)8.39(s，1H)11.13(br.s.，1H)13.17(s，1H)
4- [ (3-dimethylamino-propyl) -methyl-amino ] -N- [5- (3-ethoxy-benzoyl) -1H-indazol-3-yl ] -2-nitro-benzamide [ (II) R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- [ (3-dimethylamino-propyl) -methyl-amino ] -2-nitro-phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33(t，J=6.95Hz， 3H)1.60-1.78(m，2H)2.22(s，6H)2.29-2.37(m，2H)3.01(s，3H)3.48(t，J=7.01Hz，2H)4.09(q，J=6.99Hz，2H)6.98(dd，J=8.84，2.50Hz，1H)7.16-7.21(m，2H)7.22-7.25(m，1H)7.27-7.32(m，1H)7.45(t，J=7.93Hz，1H)7.58-7.66(m，2H)7.83(dd，J=8.78，1.59Hz，1H)8.36(s，1H)11.04(s，1H)13.14(s，1H)
Step i
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl]cpd (compound) 6
N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide (3.61g, 6.93mmol) was dissolved in DCM (150mL) under an argon atmosphere and trifluoroacetic acid (150mL) was added with stirring. Sodium borohydride particles (2.62g, 69.3mmol) were added stepwise over 72 hours. The reaction mixture was evaporated, dissolved in mixture MeOH/acetone and stirred for 1 hour. The resulting mixture was evaporated to dryness, redissolved in MeOH and NaOH8N was added to reach a basic pH. The crude product was evaporated and ice/water was added, the solid thus formed was filtered, washed with water and dried in vacuo at 80 ℃ to give 3.22g of the title compound (92% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.23(s，3H)2.42-2.47(m，4H)3.33-3.38(m，4H)4.05(s，2H)6.91-6.97(m，2H)6.97-7.05(m，1H)7.24(dd，J=8.60，1.52Hz，1H)7.27(br.s.，1H)7.41(d，J=8.66Hz，1H)7.44(br.s.，1H)7.63(s，1H)7.66-7.73(m，1H)10.81(br.s.，1H)12.70(s，1H)
The following compounds were obtained by the same procedure:
4- [ (3-dimethylamino-propyl) -methyl-amino]-N- [5- (3-ethoxy-benzyl) -1H-indazol-3-yl]-2-nitro-benzamide [ (I)A) R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- [ (3-dimethylamino-propyl) -methyl-amino]-2-nitro-phenyl]cpd.53
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.26-1.31(m，3H)1.69(t，J=6.77Hz，2H)2.19(s，6H)2.28(br.s.，2H)3.02(s，3H)3.45-3.51(m，2H)3.93-4.00(m，2H)3.96(s，2H)6.70-6.73(m，1H)6.76-6.80(m，1H)6.77(d，J=1.59Hz，1H)6.98(d，J=8.90Hz，1H)7.14-7.19(m，1H)7.19-7.23(m，2H)7.38(d，J=8.66Hz，1H)7.61(s，1H)7.67(d，J=10.00Hz，1H)10.72(br.s.，1H)12.65(s，1H)
N- {5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl]-1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)B) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.60
A mixture of N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide (130mg, 0.226mmol) and sodium borohydride (15mg, 0.39mmol) was dissolved in isopropanol (20mL) at room temperature. The reaction mixture was stirred for 4 hours, quenched with methanol and evaporated to dryness. The crude product was re-dissolved in DCM and washed with brine. After trituration with ether, 59mg of the title compound were recovered (45% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.26-1.41(m，2H)1.89-1.99(m，2H)2.23(s，3H)2.39-2.47(m，4H)3.21-3.29(m，4H)3.45-3.55(m，2H)3.63-3.74(m，1H)3.76-3.86(m，2H)5.81(d，J=4.15Hz，1H)6.12(d，J=4.15Hz，1H)6.14(d，J=2.07Hz，1H)6.24(dd，J=9.08，2.26Hz，1H)6.96-7.04(m，1H)7.05-7.12(m，2H)7.27-7.36(m，1H)7.37-7.43(m，1H)7.64(s，1H)7.80(d，J=9.15Hz，1H)8.31(d，J=7.56Hz，1H)10.09(s，1H)12.63(s，1H)
N- {5- [ (3-ethoxy-phenyl) -hydroxy-methyl]-1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (I)B) R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl]cpd.67
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29(t，J=6.95Hz，3H)2.24(s，3H)2.42-2.47(m，4H)3.36(m，4H)3.97(q，J=6.95Hz，2H)5.70(d，J=3.90Hz，1H)5.85(d，J=3.90Hz，1H)6.72(ddd，J=8.17，2.56，0.73Hz，1H)6.90(d，J=7.68Hz，1H)6.93(dd，J=2.20，1.46Hz，1H)7.17(t，J=7.87Hz，1H)7.26(d，J=8.78Hz，1H)7.28(dd，J=8.72，1.40Hz，1H)7.36(d，J=8.78Hz，1H)7.44(d，J=2.07Hz，1H)7.70(d，J=6.71Hz，1H)7.81(br.s.，1H)10.80(br.s.，1H)12.65(s，1H)。
Example 2
Step j
5- (3, 5-difluoro-benzyl) -2-fluoro-benzonitrile [ (XIV), R1= R2= R3= H, R =3, 5-difluorophenyl ]
5- [ (3, 5-difluoro-phenyl) -hydroxy-methyl ] -2-fluoro-benzonitrile (3.5g, 13.3mmol) and sodium iodide (20g, 133mmol) were stirred under a nitrogen atmosphere and in acetonitrile (50mL) at 60 ℃. Chlorotrimethylsilane (17mL, 134mmol) was added to the reaction mixture stepwise over 8 hours. The mixture was diluted with ethyl acetate and washed with water, saturated aqueous sodium bicarbonate, 10% aqueous sodium thiosulfate, and brine. The crude product was purified by silica gel chromatography (EtOAc/hexane 5: 100) to give 3.1g of the title compound (88% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.02(s，2H)7.02-7.11(m，3H)7.47(t，J=9.08Hz，1H)7.68-7.74(m，1H)7.90(dd，J=6.22，2.19Hz，1H)
The following compounds were obtained by operating in the same manner:
5-benzyl-2-fluoro-benzonitrile [ (XIV), R1= R2= R3= H, R = phenyl ]
ESI(+)MS：m/z229(MNH4 +)。
Step k
5- (3, 5-difluoro-benzyl) -2-fluoro-benzonitrile [ (XIV), R1= R2= R3= H, R =3, 5-difluorophenyl ]
To a dry flask under an argon atmosphere were added 3-cyano-4-fluorophenylboronic acid (1.649g, 10mmol), potassium phosphate powder (4.254g, 20mmol) and Pd (PPh)3)4(231mg, 0.2 mmol). The flask was evacuated and back-filled with argon three times, then toluene (30mL) and 3, 5-difluorobenzyl bromide (1.295mL, 10mmol) were added via syringe through a sieve stopper with stirring. The reaction mixture was heated to 100 ℃ and maintained at this temperature for 1.5 hours in a half hour. The black mixture was dissolved in ether (200mL), washed with saturated aqueous ammonium chloride (2 × 20mL), brine (3 × 30mL), dried over sodium sulfate and evaporated to dryness to give 3.21g of a yellow oil. The crude product was purified by flash chromatography on silica gel with n-hexane/ethyl acetate 95: 5 elution gave 1.89g (76.4% yield) of a whitish solid.
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.02(s，2H)7.02-7.11(m，3H)7.47(t，J=9.08Hz，1H)7.68-7.74(m，1H)7.90(dd，J=6.22，2.19Hz，1H)
The following compounds were prepared according to the same method:
5- (2, 5-difluoro-benzyl) -2-fluoro-benzonitrile
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.01(s，2H),7.09-7.17(m，1H)，7.20-7.27(m，2H)，7.46(t，J=9.08Hz，1H)，7.64(m,1H)，7.82(dd，J=6.22，2.19Hz，1H)
2-fluoro-5- (5-fluoro-2-methyl-benzyl) -benzonitrile
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.18(s，3H)，4.01(s，2H)，7.00(m，2H)，7.22(m，1H)，7.48(t，J=9.08Hz，1H)，7.56(m,1H)，7.75(dd，J=6.22，2.19Hz，1H)
2-fluoro-5- (3-fluoro-benzyl) -benzonitrile
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.98(s，2H)，6.95-7.15(m，3H)，7.27-7.38(m，1H)，7.38-7.48(t，1H)，7.61-7.70(m，1H)，7.81-7.87(dd，J=6.22，2.19Hz.1H)。
2-fluoro-5-pyridin-3-ylmethyl-benzonitrile
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.03(s，2H)7.33(ddd，J=7.83，4.79，0.79Hz，1H)7.47(t，J=9.02Hz，1H)7.65-7.68(m，1H)7.68-7.72(m，1H)7.89(dd，J=6.28，2.01Hz，1H)8.44(dd，J=4.76，1.59Hz，1H)8.54(d，J=1.71Hz，1H)
Step l
5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine [ (III)A) R1= R2= R3= H, R =3, 5-difluorophenyl]
A mixture of 5- (3, 5-difluoro-benzyl) -2-fluoro-benzonitrile (20g, 80.9mmol) and hydrazine hydrate (19.6mL, 404mmol) in n-butanol (200mL) was heated at 120 ℃ overnight. The reaction mixture was diluted with water/ethyl acetate and the organic phase was washed twice with brine, dried and evaporated. The crude product was triturated with ether and filtered to give 13gr final product. The resulting organic phase was purified by silica gel chromatography (DCM/EtOH 95: 5) to give 6.3g of the title compound (19.2 g total, 92% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.01(s，2H)5.23(s，2H)6.89-6.98(m，2H)7.03(tt，J=9.43，2.33Hz，1H)7.11-7.15(m，1H)7.16-7.20(m，1H)7.53(s，1H)11.30(s，1H)
The following compounds were prepared according to the same method:
5- (2, 5-difluoro-benzyl) -1H-indazol-3-ylamine
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.99(s，2H)，5.28(m，2H),7.05-7.25(m，5H)，7.51(s,1H)，11.30(bs，1H)。
5- (5-fluoro-2-methyl-benzyl) -1H-indazol-3-ylamine
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.21(s，3H)，3.97(s，2H)，5.22(bs，2H)，7.43(s，1H),7.14-7.20(m，2H)，7.06(dd，1H)，6.87-6.97(m，2H)，11.27(bs，1H)。
5- (3-fluoro-benzyl) -1H-indazol-3-ylamine
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.00(s，2H)，5.22(bs，2H),6.96-7.09(m，3H)，7.11(m，1H)，7.15(m，1H)，7.29-7.37(m，1H)，7.53(s，1H)，11.27(s，1H)。
5-pyridin-3-ylmethyl-1H-indazol-3-ylamine
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.01(s，2H)5.23(br.s.，2H)7.08-7.15(m，1H)7.15-7.19(m，1H)7.25-7.34(m，1H)7.53(s，1H)7.60(dt，J=7.86，1.92Hz，1H)8.40(dd，J=4.69，1.65Hz，1H)8.51(d，J=1.83Hz，1H)11.28(s，1H)
5-benzyl-1H-indazol-3-ylamine
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.97(s，2H)5.21(s，2H)7.07-7.11(m，1H)7.13-7.16(m，1H)7.16-7.20(m，1H)7.20-7.24(m，2H)7.25-7.31(m，2H)7.52(s，1H)11.25(s，1H)。
Step n
5- [1- (3, 5-difluoro-phenyl) -ethyl]-2-fluoro-benzonitrile [ (XIXD)1) R1= R2= R3= H, R =3, 5-difluorophenyl, R' = methyl]
5- (3, 5-difluoro-benzyl) -2-fluoro-benzonitrile (450mg, 1.82mmol) was dissolved in dry THF (14mL) under a nitrogen atmosphere and at-20 deg.C and iodomethane (0.17mL, 2.73mmol) was added with stirring. A1.0M (0.684ml, 3.64mmol) solution of lithium bis- (trimethylsilyl) -amide in THF was added stepwise. After 20 minutes, by adding KHSO4The reaction was quenched with 10% solution and extracted with ethyl acetate. With KHSO4The organic phase was washed with 10% aqueous solution and brine, dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on silica gel using hexane/ethyl acetate 98/2 as eluent. The title product was isolated as an oil (400mg, 84% yield))。
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.59(d，J=7.32Hz，3H)4.31(q，J=7.19Hz，1H)7.08(m，3H)7.46(t，J=9.15Hz，1H)7.73(m，1H)7.95(dd，J=6.22，2.44Hz，1H)
L'
5- [1- (3, 5-difluoro-phenyl) -ethyl]-1H-indazol-3-ylamine [ (IIID)1) R1= R2= R3= H, R =3, 5-difluorophenyl, R' = methyl]
5- [1- (3, 5-difluoro-phenyl) -ethyl ] -2-fluoro-benzonitrile (324mg, 1.24mmol) was dissolved in n-butanol (3mL) and hydrazine hydrate (0.301mL, 6.20mmol) was added. The reaction mixture was stirred at 120 ℃ for 22 hours and then quenched by addition of water/ethyl acetate. The organic phase was separated, washed with water and brine, dried over sodium sulfate and evaporated to dryness. The crude product was purified by silica gel chromatography eluting with a gradient of DCM/EtOH 99/1-98/2. The title product was isolated as an oil (96mg, 39% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.61(d，J=7.19Hz，3H)4.25(q，J=7.32Hz，1H)5.26(br.s，5.26，2H)6.99(m，3H)7.12(dd，J=8.66，1.59Hz，1H)7.16(dd，J=8.54，0.73Hz，1H)7.62(br.s，1H)11.29(s，1H)
Step i'
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.11
To a suspension of 4- (4-methyl-piperazin-1-yl) -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid trifluoroacetate (10g, 22.1mmol) in dry dichloromethane (300mL) was added oxalyl chloride (3.58mL, 42.3mmol) and N, N-dimethylformamide (1-2 drops). The mixture was stirred at room temperature for 2 hours and then evaporated to dryness. The resulting crude acid chloride was dissolved in toluene and evaporated again, then dissolved in dry tetrahydrofuran (130mL) at-20 ℃. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (5g.19.28mmol) and N, N-diisopropylethylamine (12.8mL, 73.3mmol) in dry THF (40mL) was added to the cooled reaction mixture. The mixture was stirred at-20 ℃ for 4 hours and then quenched by addition of water/ethyl acetate. The organic phase is washed with saturated sodium bicarbonate solution, dried over sodium sulfate and evaporated to dryness.
The crude product was purified by flash chromatography on silica gel using dichloromethane/ethanol 100: 10 as eluent, intermediate N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzamide was obtained as a light yellow solid.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.25-1.41(m，1H)1.48-1.61(m，1H)1.66(d，J=9.02Hz，1H)1.92(d，J=9.15Hz，1H)2.25(s，3H)2.43-2.49(m，4H)3.23-3.41(m，6H)3.77(dd，J=10.91，4.21Hz，1H)3.87(dd，J=11.65，3.96Hz，1H)4.02(s，2H)4.37-4.49(m，1H)6.89(d，J=2.44Hz，1H)6.90-6.98(m，2H)7.02(tt，J=9.42，2.29Hz，1H)7.09(dd，J=8.78，2.44 Hz，1H)7.27(dd，J=8.72，1.40Hz，1H)7.41-7.43(m，2H)7.83(d，J=8.78Hz，1H)10.52(s，1H)12.69(s，1H)
Alternatively, the crude reaction mixture, which had not been purified beforehand, can be dissolved in methanol (375mL) in the presence of triethylamine (60mL) and stirred at 65 ℃ for 2 hours. The solvent was removed under reduced pressure and the residue was treated with water/ethyl acetate. The organic phase was dried over sodium sulfate and evaporated to dryness. Chromatography on silica gel (DCM/EtOH/NH in MeOH)35N =1000/50/5) and the compound thus obtained was crystallized from EtOAc/hexane to give 8.4g of the title compound as a white solid (7)8% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.26-1.43(m，2H)1.86-2.02(m，2H)2.23(s，3H)2.42-2.46(m，4H)3.23-3.29(m，4H)3.45-3.54(m，2H)3.62-3.75(m，1H)3.82(dt，J=11.61，3.83Hz，2H)4.05(s，2H)6.14(d，J=2.07Hz，1H)6.24(dd，J=8.90，2.19Hz，1H)6.94-7.06(m，3H)7.26(dd，J=8.66，1.46Hz，1H)7.41(d，J=8.66Hz，1H)7.50(d，1H)7.80(d，J=9.15Hz，1H)8.29(d，J=7.68Hz，1H)10.08(s，1H)12.63(s，1H)。
The following compounds were obtained by operating in the same manner:
n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- (3-methoxy-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (3-methoxy-propylamino) -phenyl]cpd.36
1H-NMR (400MHz), (ppm, DMSO-d 6): 1.80 (quintuple, J =6.49Hz, 2H)2.24(s, 3H)2.42-2.47(m, 4H)3.16-3.21(m, 2H)3.23(s, 3H)3.26-3.32(m, 4H)3.41(t, J =6.16Hz, 2H)4.04(s, 2H)6.07(d, J =2.19Hz, 1H)6.24(dd, J =9.02, 2.19Hz, 1H)6.95-7.00(m, 2H)6.99-7.04(m, 1H)7.24(dd, J =8.66, 1.59Hz, 1H)7.41(d, J =8.54Hz, 1H)7.51(s, 1H)7.80(d, J =9.15, 1H = 8.15, 1H) 1.12 (s, 1H)7.80(d, J = 9.19H), 1H = 8.12H) 1H (s, 1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -phenyl]cpd.4
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.23(s，3H)2.44-2.49(m，4H)3.28-3.32(m，4H)4.05(s，2H)6.90-7.00(m，3H)7.02(d，J=9.15Hz，2H)7.24(dd，J=8.66，1.59Hz，1H)7.41(d，J=0.49Hz，1H)7.59(s，1H)7.97(d，J=9.02Hz，2H)10.39(s，1H)12.67(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- ((R) -2-methoxy-1-methyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- ((R) -2-methoxy-1-methyl-ethylamino) -phenyl]cpd.32
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.14(d，J=6.34Hz，3H)2.23(s，3H)2.41-2.47(m，4H)3.24-3.31(m，4H)3.27(s，3H)3.32-3.40(m，2H)3.74-3.83(m，1H)4.05(s，2H)6.13(d，J=2.19Hz，1H)6.24(dd，J=9.02，2.20Hz，1H)6.94- 7.04(m，3H)7.25(dd，J=8.66，1.59Hz，1H)7.41(d，J=8.54Hz，1H)7.49(s，1H)7.78(d，J=9.02Hz，1H)8.20(d，J=7.68Hz，1H)10.04(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- (2-methoxy-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (2-methoxy-ethylamino) -phenyl]cpd.26
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.25(s，3H)2.44-2.49(m，4H)3.26(s，3H)3.27-3.31(m，6H)3.54(t，J=5.37Hz，2H)4.05(s，2H)6.09(d，J=1.95Hz，1H)6.25(dd，J=8.96，2.01Hz，1H)6.94-7.00(m，2H)6.99-7.05(m，1H)7.24(dd，J=8.60，1.52Hz，1H)7.41(d，J=8.66Hz，1H)7.51(s，1H)7.79(d，J=9.15Hz，1H)8.23(t，J=5.12Hz，1H)10.06(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- [ (3-dimethylamino-propyl) -methyl-amino]-2-nitro-benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (3-dimethylamino-propyl) -methyl-amino]-2-nitro-phenyl]cpd.59
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.68(m，2H)2.15(m， 6H)2.25(t，J=6.58Hz，2H)3.02(s，3H)3.48(t，J=7.07Hz，2H)4.05(s，2H)6.93-7.05(m，4H)7.19(d，J=2.44Hz，1H)7.26(dd，J=8.54，1.46Hz，1H)7.42(d，J=8.54Hz，1H)7.62(s，1H)7.68(bs，1H)10.73(s，1H)12.69(s，1H)
2-cyclohexylamino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-cyclohexylamino-phenyl]cpd.18
1H-NMR(400MHz)，(ppm，DMSO-d6)：12.61(s，1H)10.04(s，1H)8.26(d，1H)7.77(d，1H)7.48(s，1H)7.40(d，1H)7.25(dd，1H)6.90-7.00(m，3H)6.21(dd，1H)6.08(d，1H)4.03(s，2H)3.45(m，1H)3.25(m，4H)2.45(bs，4H)2.24(s，3H)1.88-1.23(m，10H)
N- {5- [1- (3, 5-difluoro-phenyl) -ethyl ] -1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (ID), R1= R2= R3= R "= H, R =3, 5-difluorophenyl, R' = methyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl ] cpd.75
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.43(m，2H)1.60(d，J=7.19Hz，3H)1.89-1.99(m，2H)2.29(br.s.，3H)2.45-2.57(m，4H)3.22-3.38(m，4H)3.45-3.55(m，2H)3.64-3.76(m，1H)3.78-3.85(m，2H)4.31(q，J=7.40Hz，1H)6.15 (d，J=1.95Hz，1H)6.25(dd，J=8.90，2.19Hz，1H)6.94-7.06(m，3H)7.28(dd，J=8.78，1.59Hz，1H)7.40(d，J=8.54Hz，1H)7.52(s，1H)7.81(d，J=9.15Hz，1H)8.32(d，J=7.68Hz，1H)10.09(s，1H)12.62(s，1H)
By preparative chiral-HPLC using Daicel Chiralpak AD250 × 20mm10 μm as column system, hexane/2-propanol 40: 60 as eluent, the single enantiomer was obtained.
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (1-methoxy-2-methylpropan-2-yl) amino]-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (1-methoxy-2-methylpropan-2-yl) amino]-phenyl radical]cpd.34
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31(s，6H)2.27(br.s.，3H)2.50(m，4H)3.26(m，7H)3.35(s，2H)4.05(s，2H)6.27(dd，J=9.02，2.32Hz，1H)6.31(d，J=2.32Hz，1H)6.93-7.05(m，3H)7.25(dd，J=8.60，1.52Hz，1H)7.41(d，J=8.54Hz，1H)7.51-7.53(m，1H)7.76(d，J=8.90Hz，1H)8.26(s，1H)10.14(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- (2-methoxy-1-methoxymethyl-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (2-methoxy-1-methoxymethyl-ethylamino) -phenyl]cpd.16
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.42(br.s.，3H)2.70(br.s.，4H)3.26(s，6H)3.30(m，4H)3.41(d，J=5.00Hz，4H)3.85(m，J=8.17，5.00，5.00，5.00，5.00Hz，1H)4.04(s，2H)6.20(d，J=1.95Hz，1H)6.26(dd，J=8.96，2.01Hz，1H)6.94-7.04(m，3H)7.24(dd，J=8.66，1.46Hz，1H)7.41(d，J=8.54Hz，1H)7.48(br.s.，1H)7.79(d，J=9.02Hz，1H)8.32(d，J=8.29Hz，1H)10.06(s，1H)12.64(s，1H)
2-benzylamino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-benzylamino-phenyl]cpd.24
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.22(s，3H)2.41(br.s.，4H)3.19-3.24(m，4H)4.04(s，2H)4.39(d，J=5.49Hz，2H)6.09(d，J=2.19Hz，1H)6.26(dd，J=9.02，2.32Hz，1H)6.92-6.98(m，2H)6.98-7.04(m，1H)7.21-7.27(m，2H)7.30-7.36(m，2H)7.36-7.39(m，2H)7.40(d，J=9.02Hz，1H)7.51(s，1H)7.81(d，J=9.02Hz，1H)8.60(t，J=5.55Hz，1H)10.11(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- (2-fluoro-ethylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (2-fluoro-ethylamino) -phenyl]cpd. 38
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.24(s，3H)2.43-2.48(m，4H)3.26-3.31(m，4H)3.49(dq，J=27.68，5.12Hz，2H)4.04(s，2H)4.60(dt，J=47.68，4.76Hz，2H)6.12(d，J=2.23Hz，1H)6.28(dd，J=8.99，2.23Hz，1H)6.94-7.00(m，2H)6.99-7.04(m，1H)7.24(dd，J=8.57，1.52Hz，1H)7.41(d，J=8.57Hz，1H)7.51(s，1H)7.81(d，J=8.99Hz，1H)8.37(t，J=5.43Hz，1H)10.11(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2- (2-fluoro-propylamino) -4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (2-fluoro-propylamino) -phenyl]cpd.40
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.86-2.04(m，2H)2.26(br.s.，3H)2.48(br.s.，4H)3.21-3.37(m，6H)4.04(s，2H)4.44-4.66(dt，J=47.43，5.73Hz，2H)6.09(d，J=1.95Hz，1H)6.26(dd，J=9.02，2.20Hz，1H)6.94-7.05(m，3H)7.25(dd，J=8.60，1.40Hz，1H)7.41(d，J=8.66Hz，1H)7.50(d，J=1.71Hz，1H)7.81(d，J=9.02Hz，1H)8.22(t，J=5.24Hz，1H)10.09(s，1H)12.63(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- [ (3-dimethylamino-propyl) -methyl-amino]-2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (3-dimethylamino-propyl) -methyl-amino]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.55
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.32-1.44(m，2H)1.67(quin，J=6.98Hz，2H)1.93-1.98(m，2H)2.17(s，6H)2.26(t，J=6.65Hz，2H)2.96(s，3H)3.36-3.43(m，2H)3.44-3.53(m，2H)3.58-3.69(m，1H)3.79-3.87(m，2H)4.05(s，2H)5.87(d，J=2.19Hz，1H)6.04(dd，J=9.02，2.32Hz，1H)6.96-7.05(m，3H)7.25(dd，J=8.60，1.52Hz，1H)7.41(d，J=8.54Hz，1H)7.49(s，1H)7.77(d，J=9.15Hz，1H)8.35(d，J=7.32Hz，1H)9.96(s，1H)12.60(s，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-phenylamino-benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-phenylamino-phenyl]cpd.42
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.24(s，3H)2.46(br.s.，4H)3.22(br.s.，4H)4.05(s，2H)6.53(dd，J=9.02，2.19Hz，1H)6.74(d，J=2.32Hz，1H)6.95-7.02(m，4H)7.19(d，J=7.56Hz，2H)7.25(dd，J=8.66，1.46Hz，1H)7.29-7.35(m，2H)7.40-7.44(m，1H)7.55(s，1H)7.91(d，J=9.15Hz，1H) 10.03(s，1H)10.39(s，1H)12.69(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-methyl-1, 4-diazepan-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-1, 4-diazepan-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.89
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.42(m，2H)1.83-1.98(m，4H)2.28(s，3H)2.44-2.49(m，2H)2.63(d，J=4.51Hz，2H)3.44-3.59(m，6H)3.65(d，J=11.46Hz，1H)3.78-3.85(m，2H)4.04(s，2H)5.87(d，J=2.32Hz，1H)6.05(dd，J=9.08，2.26Hz，1H)6.96-7.04(m，3H)7.25(dd，J=8.59，1.52Hz，1H)7.41(d，J=8.53Hz，1H)7.49(s，1H)7.77(d，J=9.14Hz，1H)8.36(d，J=7.68Hz，1H)9.96(s，1H)12.60(s，1H)
N- [5- (3, 5-difluoro-)Benzyl) -1H-indazol-3-yl]-4- [ (2-dimethylamino-ethyl) -methyl-amino]-2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (2-dimethylamino-ethyl) -methyl-amino]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.90
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.32-1.43(m，2H)1.96(d，1H)2.19-2.22(m，6H)2.40(t，J=7.19Hz，2H)2.98(s，3H)3.41-3.51(m，4H)3.56-3.65(m，1H)3.80-3.87(m，2H)4.04(s，2H)5.87(d，J=2.32Hz，1H)6.02(dd，J=9.08，2.38Hz，1H)6.96-7.04(m，3H)7.25(dd，J=8.59，1.52Hz，1H)7.41(d，J=8.53Hz，1H)7.49(s，1H)7.78(d，J=9.14Hz，1H)8.35(d，J=7.31Hz，1H)9.97(s，1H)12.60(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [4- (dimethylamino) piperidin-1-yl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [4- (dimethylamino) piperidin-1-yl group]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.91
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.43(m，4H)1.82(d，J=12.32Hz，2H)1.93(dq，J=12.74，2.77Hz，2H)2.20(s，6H)2.29(m，1H)2.78(td，J=12.38，2.19Hz，2H)3.49(ddd，J=11.86，9.91，2.26Hz，2H)3.62-3.72(m，1H)3.81(dt，J=11.74，4.07Hz，2H)3.87(d，J=12.56Hz，2H)4.04(s，2H)6.12(d，J=2.19Hz，1H)6.23(dd，J=8.96，2.26Hz，1H)6.99(m，3H)7.25(dd，J=8.60，1.52Hz，1H)7.40(d，J=8.54Hz，1H)7.48(br.s.，1H)7.78(d，J=9.15Hz，1H)8.28(d，J=7.56Hz，1H)10.05(s，1H)12.61(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.92
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.36(m，2H)1.72(m，4H)1.99(m，6H)2.43(m，3H)2.63(m，2H)3.16(m，2H)3.39-3.47(m，3H)3.58(br.s.，1H)3.82-3.90(m，2H)3.90(br.s.，1H)4.04(s，2H)5.82(d，J=1.59Hz，1H)5.90(dd，J=8.90，2.07Hz，1H)6.98(m，3H)7.24(dd，J=8.60，1.52Hz，1H)7.40(d，J=8.90Hz，1H)7.48(br.s.，1H)7.77(d，J=9.02Hz，1H)8.36(d，J=7.32Hz，1H)9.95(s，1H)12.60(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-3- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =3- (4-methylpiperazin-1-yl) phenyl]cpd.93
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.26(s，3H)2.47-2.54(m，4H)3.22-3.27(m，4H)4.06(s，2H)6.92-6.99(m，2H)6.99-7.06(m，1H)7.15-7.20(m，1H)7.26(dd，J=8.66，1.59Hz，1H)7.36(t，J=7.93Hz，1H)7.42-7.45(m，1H)7.47(d，J=7.80Hz，1H)7.60-7.63(m，2H)10.65(s，1H)12.73(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (piperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (piperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl]cpd.98
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.41(m，2H)1.88-2.01(m，2H)2.81-2.88(m，4H)3.17-3.22(m，4H)3.45-3.54(m，2H)3.62-3.73(m，1H)3.78-3.85(m，2H)4.05(s，2H)6.12(d，J=2.19Hz，1H)6.23(dd，J=8.96，2.26Hz，1H)6.94-7.04(m，3H)7.26(dd，J=8.65，1.58Hz，1H)7.39-7.43(m，1H)7.49(s，1H)7.80(d，J=9.02Hz，1H)8.29(d，J=7.68Hz，1H)10.07(s，1H)12.63(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-Methylpiperazin-1-yl) -2- { [ cis-4- (trifluoromethyl) cyclohexyl]Amino benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- { [ cis-4- (trifluoromethyl) cyclohexyl]Amino } phenyl]cpd.99
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.39-1.53(m，2H)1.58-1.73(m，4H)1.84-1.91(m，2H)2.25(s，3H)2.28-2.40(m，1H)2.47(br.s.，4H)3.25-3.33(m，4H)3.82-3.90(m，1H)4.01(s，2H)6.10(d，J=1.95Hz，1H)6.24(dd，J=9.15，2.19Hz，1H)6.90-6.96(m，2H)6.96-7.03(m，1H)7.24(dd，J=8.60，1.52Hz，1H)7.42(d，J=8.54Hz，1H)7.52(s，1H)7.83(d，J=9.02Hz，1H)8.69(d，J=7.80Hz，1H)10.10(s，1H)12.65(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-Methylpiperazin-1-yl) -2- { [ trans-4- (trifluoromethyl) cyclohexyl]Amino benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- { [ trans-4- (trifluoromethyl) cyclohexyl]Amino } phenyl]cpd.100
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.10-1.23(m，2H)1.44-1.57(m，2H)1.86-1.94(m，2H)2.06-2.15(m，2H)2.25(s，3H)2.29-2.34(m，1H)2.46(br.s.，4H)3.24-3.31(m，4H)3.39-3.51(m，1H)4.05(s，2H)6.15(d，J=2.07Hz，1H)6.23(dd，J=8.90，2.07Hz，1H)6.95-7.00(m，2H)7.00-7.06(m，1H)7.25(dd，J=8.60，1.52Hz，1H)7.41(d，J=8.54Hz，1H)7.48(s，1H)7.78(d，J=9.02Hz，1H)8.14(d，J=8.05Hz，1H)10.05(s，1H)12.62(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2-fluoro-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-fluoro-phenyl]cpd.101
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.26(s，3H)2.45-2.50(m，4H)3.29-3.36(m，4H)4.06(s，2H)6.78-6.89(m，2H)6.94-6.98(m，2H)6.98-7.06(m，1H)7.25(dd，J=8.54，1.59Hz，1H)7.42(d，J=8.66Hz，1H)7.64(s，1H)7.68(t， J=8.90Hz，1H)10.08(d，J=3.41Hz，1H)12.68(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (1-methylpiperidin-4-yl) amino]Benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =2- [ (1-methylpiperidin-4-yl) amino]-phenyl radical]cpd.110
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.35-1.49(m，H)1.94(d，H)2.20(br.s.，5H)2.60-2.73(m，2H)3.38-3.47(m，1H)4.05(s，2H)6.58-6.64(m，1H)6.80(d，J=8.29Hz，1H)6.95-7.00(m，2H)7.00-7.05(m，1H)7.27(dd，J=8.65，1.58Hz，1H)7.32-7.37(m，1H)7.44(d，J=8.53Hz，1H)7.53(s，1H)7.85-7.88(m，1H)7.89(dd，J=8.05，1.34Hz，1H)10.44(s，1H)12.72(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (1-methylpiperidin-4-yl) amino]-4- (morpholin-4-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (morpholin-4-yl) -2- [ (1-methylpiperidin-4-yl) amino]-phenyl radical]cpd.111
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.38-1.50(m，2H)1.91-2.01(m，2H)2.27(m，5H)2.72(m，2H)3.20-3.26(m，4H)3.50(br.s.，1H)3.72-3.78(m，4H)4.05(s，2H)6.11(d，J=2.19Hz，1H)6.25(dd，J=9.08，2.13Hz，1H)6.92-7.08(m，3H)7.26(dd，J=8.59，1.52Hz，1H)7.42(d，J=8.53Hz，1H)7.50 (s，1H)7.82(d，J=9.02Hz，1H)8.28(d，J=7.31Hz，1H)10.10(s，1H)12.64(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2-methoxy-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-methoxy-amino } phenyl]cpd.112
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.25(s，3H)2.47(br.s.，4H)3.33-3.38(m，4H)4.02(s，3H)4.06(s，2H)6.63(d，J=1.95Hz，1H)6.67(dd，J=8.96，2.13Hz，1H)6.96(dd，J=8.72，2.13Hz，2H)6.99-7.05(m，1H)7.24(dd，J=8.66，1.59Hz，1H)7.40(d，J=8.66Hz，1H)7.76(s，1H)7.88(d，J=8.78Hz，1H)9.99(s，1H)12.65(s，1H)
N- [5- (2, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R =2, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.10
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.42(m，2H)1.90-1.98(m，2H)2.27(br.s.，3H)2.49(br.s.，4H)3.24-3.32(m，4H)3.45-3.56(m，2H)3.64-3.74(m，1H)3.82(ddd，J=11.80，3.96，3.75Hz，2H)4.04(s，2H)6.14(d，J=1.83Hz，1H)6.24(dd，J=8.90，1.95Hz，1H)7.04-7.12(m，1H)7.15- 7.23(m，2H)7.24-7.27(m，1H)7.41(d，J=8.66Hz，1H)7.46(s，1H)7.80(d，J=9.02Hz，1H)8.30(d，J=7.68Hz，1H)10.08(s，1H)12.63(s，1H)
N- [5- (2-methyl-5-fluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R = 2-methyl-5-fluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.135
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.41(m，2H)1.94(dd，J=13.35，2.86Hz，2H)2.22(s，3H)2.25(s，3H)2.46(br.s.，4H)3.24-3.30(m，4H)3.46-3.54(m，2H)3.63-3.73(m，1H)3.78-3.86(m，2H)4.03(s，2H)6.13(d，J=1.95Hz，1H)6.23(dd，J=9.02，2.07Hz，1H)6.89-6.98(m，2H)7.14-7.21(m，2H)7.38(s，1H)7.41(d，J=8.65Hz，1H)7.79(d，J=9.02Hz，1H)8.31(d，J=7.80Hz，1H)10.07(s，1H)12.61(s，1H)
N- [5- (2-fluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R = 2-fluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.9
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.42(m，2H)1.89-1.98(m，2H)2.24(s，3H)2.45(br.s.，4H)3.24-3.30(m，4H)3.47-3.55(m，2H)3.64-3.74(m，1H)3.77-3.87(m，2H)4.04(s，2H)6.14(d，J=2.19Hz，1H)6.24(dd，J=8.96，2.26Hz，1H)6.95-7.02(m，1H)7.04-7.09(m，1H)7.10(d，J=7.56Hz，1H)7.24(dd，J=8.66，1.46Hz，1H)7.31(td，J=7.80，6.34Hz，1H)7.40(d，J=8.53Hz，1H)7.46(s，1H)7.79(d，J=9.02Hz，1H)8.28(d，J=7.80Hz，1H)10.07(s，1H)12.61(s，1H)
4- (4-methylpiperazin-1-yl) -N- [5- (pyridin-3-ylmethyl) -1H-indazol-3-yl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R = pyridin-3-yl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.136
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.42(m，2H)1.95(d，2H)2.26(s，3H)2.47(br.s.，4H)3.25-3.30(m，4H)3.47-3.54(m，2H)3.64-3.74(m，1H)3.79-3.86(m，2H)4.05(s，2H)6.14(d，J=2.07Hz，1H)6.24(dd，J=8.90，2.19Hz，1H)7.24(dd，J=8.60，1.52Hz，1H)7.29(ddd，J=7.80，4.76，0.73Hz，1H)7.41(d，J=8.90Hz，1H)7.47(s，1H)7.63(dt，J=7.87，1.92Hz，1H)7.79(d，J=9.15Hz，1H)8.27(d，J=7.80Hz，1H)8.39(dd，J=4.76，1.59Hz，1H)8.52(d，J=1.71Hz，1H)10.07(s，1H)12.62(s，1H)
N- [ 5-benzyl-1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R = phenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.137
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.42(m，2H)1.90-1.99(m，2H)2.24(s，3H)2.42-2.47(m，4H)3.24-3.31(m，4H)3.46-3.55(m，2H)3.64-3.76(m，1H)3.78-3.87(m，2H)4.01(s，2H)6.14(d，J=2.07Hz，1H)6.24(dd，J=8.96，2.26Hz，1H)7.17-7.27(m，6H)7.38(d，J=8.90Hz，1H)7.44(s，1H)7.79(d，J=9.02Hz，1H)8.28(d，J=7.68Hz，1H)10.05(s，1H)12.59(s，1H)。
Example 3
Step r
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (XXIV)A) R1= R2= R3= H, R =3, 5-difluorophenyl, PG1= trifluoroacetyl group]
To a suspension of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (0.5g, 1.93mmol) in anhydrous dichloromethane (20mL) with vigorous stirring and cooling to 0 ℃ was added dropwise trifluoroacetic anhydride and the thick slurry was stirred for 3.5 hours. The reaction mixture was poured into 3% NaHCO3The solution was extracted with dichloromethane. The organic layer was washed with brine, Na2SO4Dried and concentrated to give a crude white solid which was used directly in the next step.
ESI(+)MS m/z356(100，MH+) (ii) a HRMS (ESI) vs C16H10F5N3O+H+The calculated value of (a): 356.0817, respectively; measured value: 356.0820
Step s
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide [ (XXV)A) R1= R2= R3= H, R =3, 5-difluorophenyl, PG = triphenylmethyl, PG1= trifluoroacetyl group]
Mixing crude N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2,2, 2-trifluoro-acetamide was suspended in dichloromethane (25mL) and treated with trityl chloride (0.72g, 2.58mmol) under stirringAnd (6) processing. The suspension is cooled to 0 ℃ and pure 1, 8-diazabicyclo [5.4.0] is added]Undec-7-ene (0.42mL, 2.78mmol) resulted in solubilization of the intermediate. After stirring at 0 ℃ for 3 hours, the reaction mixture was poured into 50mL of ice containing 1N HCl (5mL) and extracted with dichloromethane. With NaHCO3The organic layer was washed with brine, dried and concentrated to give the crude material, which was purified by flash chromatography (eluent: DCM). The desired product was obtained as a white solid (450mg, 40% in two steps).
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.98(s，2H)6.34(d，J=8.78Hz，1H)6.93-7.07(m，4H)7.16-7.20(m，6H)7.25-7.39(m，9H)7.52(s，1H)11.99(s，1H)
Step t
5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-ylamine [ (XXVI)A) R1= R2= R3= H, R =3, 5-difluorophenyl, PG = triphenylmethyl]
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] -2,2, 2-trifluoro-acetamide (450mg, 0.75mmol) was dissolved in methanol (6mL) and triethylamine (1.5mL), and the solution was refluxed for 3 hours. The solvent was removed under reduced pressure and the residue was purified by flash chromatography (eluent: DCM). The title compound was obtained as a white foam (300mg, yield 80%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.91(s，2H)5.53(s，2H)6.24(d，J=8.78Hz，1H)6.87-6.93(m，3H)6.97-7.07(m，1H)7.17-7.23(m，3H)7.27(t，J=7.50Hz，6H)7.34(d，J=1.59Hz，6H)7.48(s，1H)
Step u
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide [ (XXII)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl, PG = triphenylmethyl]
4- (4-methyl-piperazin-1-yl) -2-nitro-benzoic acid (150mg, 0.5mmol) was suspended in anhydrous dichloromethane (10mL), 1 drop of DMF was added followed by oxalyl chloride (0.2mL, 2 mmol). After stirring at room temperature for 2 hours, the mixture was thoroughly dried under reduced pressure to give the acid chloride as a white powder. 120mg of acid chloride (0.4mmol) was dissolved in anhydrous tetrahydrofuran (3mL) and 5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-ylamine (200mg, 0.4mmol) was added. The resulting solution was cooled to 0 ℃ with stirring. After addition of diisopropylethylamine (0.2mL, 1.2mmol), the reaction mixture was stirred for 18 hours while the temperature was gradually raised from 0 ℃ to room temperature. After evaporation of the volatiles, the crude product was purified by flash chromatography (eluent: DCM/MeOH 10: 1). The title compound was isolated as a pale yellow solid (200mg, yield 67%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.24(s，3H)2.45(br.s.，4H)3.31-3.39(m，4H)3.96(s，2H)6.29(br.s.，1H)6.98(m，4H)7.29(m，17H)7.58(s，1H)7.70(br.s.，1H)10.96(br.s.，1H)
Step i'
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide hydrochloride [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-nitro-phenyl]cpd.6
To N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide (28.5mg, 0.04mmol) in bisTo a solution in an alkane (1mL) was added a di4M HCl in alkane (0.1mL), and the mixture was stirred at room temperature for 1 hour. After concentration, the residue was suspended in ether/MeOH 1: 1, stirring for 20 minutes, filtering, and mixing with the same solvent mixtureWashed and dried. The desired product was obtained as the hydrochloride derivative (19mg, 87%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.85(d，J=4.02Hz，3H)3.08-3.31(m，4H)3.53(d，J=11.71Hz，2H)4.06(s，2H)4.13(d，J=13.17Hz，2H)6.91-6.99(m，2H)6.99-7.08(m，1H)7.26(dd，J=8.54，1.34Hz，1H)7.37(d，J=6.95Hz， 1H)7.43(d，J=8.66Hz，1H)7.58(br.s.，1H)7.64(s，1H)7.78(d，J=7.44Hz，1H)10.39(br.s.，1H)10.91(br.s.，1H)12.74(br.s.，1H)
The following compounds were obtained by operating in the same manner:
2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide hydrochloride [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-amino-phenyl]cpd.8
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.84(d，J=4.39Hz，3H)3.05-3.20(m，4H)3.44-3.53(m，2H)3.85-3.94(m，2H)4.05(s，2H)6.30(d，J=1.95Hz，1H)6.36(dd，J=8.96，2.13Hz，1H)6.93-7.00(m，2H)6.99-7.05(m，1H)7.24(dd，J=8.66，1.46Hz，1H)7.41(d，J=8.41Hz，1H)7.53(s，1H)7.81(d，J=9.02Hz，1H)10.11(br.s.，1H)10.37(br.s.，1H)12.66(br.s.，1H)
Example 4
Transformation 1
2-amino-N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide [ (XXII)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-amino-phenyl, PG = triphenylmethyl]
Reacting N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]A mixture of-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide (170mg, 0.236mmol), 10% Pd-C (10mg) and ammonium formate (25mg, 0.4mmol) in methanol (5mL) was stirred at room temperature for 18 hours. The catalyst was filtered off and the solution was concentrated. The residue was dissolved in dichloromethane and taken up with NaHCO3The aqueous solution was washed, dried and concentrated to give the title compound (145mg, 87%).
ESI (+) MS m/z243(100, trityl)+)，719(16，MH+) (ii) a HRMS (ESI) vs C45H40F2N6O+H+The calculated value of (a): 719.3304, respectively; measured value: 719.3309
The following compounds were obtained by operating in the same manner:
2-amino-N- [5- (3-ethoxy-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide [ (II), R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-amino-phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33(t，J=6.95Hz，3H)2.22(s，3H)2.40-2.45(m，4H)3.16-3.22(m，4H)4.09(q，J=6.95Hz，2H)6.17(d，J=2.44Hz，1H)6.23(dd，J=9.02，2.44Hz，1H)6.53(s，2H)7.18(ddd，J=8.23，2.62，0.85Hz，1H)7.24(dd，J=2.44，1.46Hz，1H)7.29(dt，J=7.68，1.10Hz，1H)7.44(t，J=7.93Hz，1H)7.59(dd，J=8.84，0.55Hz，1H)7.71(d，J=9.15Hz，1H)7.83(dd，J=8.78，1.59Hz，1H)8.20(br.s.，1H)10.30(s，1H)13.05(s，1H)
2-amino-N- [5- (3, 5-difluoro-benzoyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -benzamide [ (II), R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-amino-phenyl ]
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.30(br.s.，3H)2.56(m，4H)3.22(m，4H)6.18(d，J=2.32Hz，1H)6.24(dd，J=9.08，2.38Hz，1H)6.57(s，2H)7.42(m，2H)7.54(tt，J=9.15，2.38Hz，1H)7.61(dd，J=8.90，0.61Hz，1H)7.73(d，J=9.02Hz，1H)7.83(dd，J=8.84，1.65Hz，1H)8.26(d，J=0.98Hz，1H)10.36(s，1H)13.11(s，1H)
Example 5
Conversion 2+ step i "
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2-methanesulfonylamino-4- (4-methyl-piperazin-1-yl) -benzamide hydrochloride [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-methanesulfonylamino-phenyl]cpd.48
To 2-amino-N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl]To a solution of-4- (4-methyl-piperazin-1-yl) -benzamide (29mg, 0.04mmol) in anhydrous dichloromethane (2mL) and dry pyridine (0.05mL) was added methanesulfonyl chloride (14.7mg, 0.01mL, 0.13mmol) and the reaction mixture was stirred at room temperature for 8 hours. The mixture was poured into ice and extracted with dichloromethane. The organic layer was washed with 0.1N HCl, then water, and anhydrous Na2SO4Drying and concentrating to obtain a crude whitish solid, which is suspended in waterAlkane (1 mL). Is added in4M HCl in an alkane (0.1mL), then the suspension was stirred overnight. After concentration, the residue was suspended in ether/MeOH 1: 1, stirred for 20 minutes, filtered, washed with the same solvent mixture and dried. The desired product was obtained as the hydrochloride salt (15mg, 0.025mmol, 63%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.86(d，J=4.27Hz，3H)3.17(s，3H)3.14-3.27(m，4H)3.55(m，2H)4.02(m，2H)4.06(s，2H)6.92(dd，J=9.02，2.32Hz，1H)6.97-7.03(m，3H)7.05(d，J=2.56Hz，1H)7.28(dd，J=8.66，1.46Hz，1H)7.45(d，J=8.54Hz，1H)7.55(s，1H)8.12(d，J=9.15Hz，1H)10.56(s，1H)10.76(s，1H)11.42(s，1H)12.84(s，1H)
The following compounds were obtained by operating in the same manner:
n- [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl]-1H-pyrrole-2-carboxamide hydrochloride [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methylpiperazin-1-yl) -2- (1H-pyrrole-2-carbamoyl) -phenyl]cpd.44
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.74(br.s.，3H)4.04(s，2H)6.09(dt，J=3.60，2.41Hz，1H)6.65(dt，J=3.87，1.78Hz，1H)6.82(dd，J=9.02，2.32Hz，1H)6.97(m，4H)7.28(dd，J=8.66，1.46Hz，1H)7.46(d，J=8.66Hz，1H)7.59(br.s.，1H)8.09(d，J=9.15Hz，1H)8.38(d，J=2.44Hz，1H)9.99(br.s.，1H)10.71(s，1H)11.69(br.s.，1H)12.51(s，1H)12.82(s，1H)
Example 6
Transformation 4
2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2-amino-phenyl]cpd.8
Reacting N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2-nitro-benzamide (3.21g, 6.33mmol), cyclohexene (20mL), bisA mixture of an alkane (200mL) and 10% Pd/C (0.8g) was stirred at 100 ℃ for 2 hours. The reaction mixture was filtered through a pad of celite, washing well with THF and MeOH. After evaporation of the organic phase, the crude product was purified by silica gel chromatography (DCM/MeOH95/5) to yield 2.51g of the title compound (83% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.23(s，3H)2.44(br. s.，4H)3.20(t，J=4.76Hz，4H)4.04(s，2H)6.18(d，J=2.44Hz，1H)6.24(dd，J=8.96，2.38Hz，1H)6.53(s，2H)6.97(m，3H)7.22(dd，J=8.66，1.59Hz，1H)7.39(d，J=8.66Hz，1H)7.52(br.s.，1H)7.72(d，J=9.02Hz，1H)10.01(s，1H)12.60(s，1H)
The following compounds were obtained by operating in the same manner:
2-amino-4- [ (3-dimethylamino-propyl) -methyl-amino]-N- [5- (3-ethoxy-benzyl) -1H-indazol-3-yl]-benzamide [ (I)A) R1= R2= R3= H, R = 3-ethoxyphenyl, Ar =4- [ (3-dimethylamino-propyl) -methyl-amino]-2-amino-phenyl]cpd.54
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.27(t，J=6.95Hz，3H)1.67(d，J=7.19Hz，2H)2.19(s，6H)2.28(t，J=6.04Hz，2H)2.90(s，3H)3.24-3.40(m，2H)3.96(q，J=6.95Hz，2H)3.95(s，2H)5.94(d，J=2.56Hz，1H)6.04(dd，J=9.02，2.56Hz，1H)6.52(s，2H)6.68-6.72(m，1H)6.76-6.79(m，1H)6.77(s，1H)7.13-7.17(m，1H)7.18(dd，J=8.60，1.65Hz，1H)7.33-7.38(m，1H)7.47(s，1H)7.69(d，J=9.02Hz，1H)9.88(s，1H)12.53(s，1H)
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } benzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.29-3.48(m，2H)4.06(s，2H)4.29(br.s.，1H)6.63(br.s.，3H)6.85(br.s.，1H)6.92-7.05(m，3H)7.26(dd，J=8.66，1.34Hz，1H)7.43(d，J=8.54Hz，1H)7.57(s，1H)7.85(d，J=8.17Hz，1H)10.46(s，1H)12.72(s，1H)
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] carbonyl } benzamide
ESI(+)MS：m/z559(MH+)。
2-amino-N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]-N4-methylbenzene-1, 4-dicarboxamide
ESI(+)MS：m/z507(MH+)。
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [4- (prop-2-yl) piperazin-1-yl ] carbonyl } benzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：0.99(d，J=6.46Hz，6H)2.43(m，4H)2.70(d，1H)3.58(m，4H)4.06(s，2H)6.54(dd，J=8.05，1.46Hz，1H)6.65(s，2H)6.75(d，J=1.46Hz，1H)6.92-7.01(m，2H)6.99-7.05(m，1H)7.26(dd，J=8.59，1.52Hz，1H)7.43(d，J=8.65Hz，1H)7.57(s，1H)7.85(d，J=8.17Hz，1H)10.45(s，1H)12.71(s，1H)
2-amino-N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]Benzene-1, 4-dicarboxamides
ESI(+)MS：m/z493(MH+)。
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (4-methylpiperazin-1-yl) carbonyl ] benzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.06(s，2H)6.60(d，J=8.29Hz，1H)6.68(s，2H)6.80(d，J=1.46Hz，1H)6.93-7.00(m，2H)7.00-7.06(m，1H)7.26(dd，J=8.53，1.58Hz，1H)7.44(d，J=8.65Hz，1H)7.55(s，1H)7.88(d，J=8.17Hz，1H)10.48(s，1H)12.73(s，1H)
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [4- (dimethylamino) piperidin-1-yl ] carbonyl } benzamide
ESI(+)MS：m/z533(MH+)。
2-amino-N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- (1-methylpiperidin-4-yl) benzene-1, 4-dicarboxamide
ESI(+)MS：m/z519(MH+)。
Example 7
Transformation 6
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.11
To 2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]To a solution of-4- (4-methyl-piperazin-1-yl) -benzamide (1.9g, 3.98mmol) in dichloromethane (80mL) were added tetrahydro-pyran-4-one (0.55mL, 5.98mmol), trifluoroacetic acid (4mL) and tetramethylammonium triacetoxyborohydride (1.57g, 5.98 mmol). The mixture was stirred at room temperature overnight, and then tetramethylammonium triacetoxyborohydride (1.57g) was added. After stirring at room temperature for a further 3 hours, the mixture is diluted with dichloromethane, washed with 2N sodium hydroxide and brine, dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on silica gel using dichloromethane/methanol/NH in MeOH35N (96: 4: 0.5) as eluent, 1.61g of the title compound was obtained (72% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.26-1.43(m，2H)1.86-2.02(m，2H)2.23(s，3H)2.42-2.46(m，4H)3.23-3.29(m，4H)3.45-3.54(m，2H)3.62-3.75(m，1H)3.82(dt，J=11.61，3.83Hz，2H)4.05(s，2H)6.14(d，J=2.07Hz，1H)6.24(dd，J=8.90，2.19Hz，1H)6.94-7.06(m，3H)7.26(dd，J=8.66，1.46Hz，1H)7.41(d，J=8.66Hz，1H)7.50(d，1H)7.80(d，J=9.15Hz，1H)8.29(d，J=7.68Hz，1H)10.08(s，1H)12.63(s，1H)
N- {5- [ (3, 5-difluoro-phenyl) -methoxy-methyl]-1H-indazol-3-yl } -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide [ (I)C) R1= R2= R3= H, R =3, 5-difluorophenyl, R' = methyl, Ar =4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.68
The title compound was isolated as a byproduct (about 15%) during preparative HPLC purification of the mixed fractions purified from column chromatography in the preparation of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide reported above.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.44(m，2H)1.91-2.01(m，2H)2.79(br.s.，3H)3.32(m，11H)3.45-3.56(m，2H)3.68-3.78(m，1H)3.80-3.88(m，2H)5.48(s，1H)6.22(d，J=2.07Hz，1H)6.30(d，J=9.02Hz，1H)7.04-7.12(m，3H)7.32(dd，J=8.78，1.46Hz，1H)7.45(d，J=8.90Hz，1H)7.64(s，1H)7.86(d，J=9.02Hz，1H)8.35(d，J=7.80Hz，1H)10.20(s，1H)12.73(s，1H)
The following compounds were obtained by operating in the same manner:
2- [ (2- { [ tert-butyl (dimethyl) silyl ] oxy } ethyl) amino ] -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (4-methylpiperazin-1-yl) benzamide
ESI(+)MS：m/z635(MH+)。
3- ({ [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl ] amino } methyl) azetidine-1-carboxylic acid tert-butyl ester
ESI(+)MS：m/z646(MH+)。
1- [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) benzyl]Piperidine trifluoroacetate salt [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = 4-piperidin-1-ylmethyl-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.115
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.32-1.46(m，3H)1.65-1.93(m，5H)1.96-2.04(m，2H)2.81-2.96(m，2H)3.32(br.s.，2H)3.44-3.54(m，2H)3.63-3.74(m，1H)3.82-3.91(m，2H)4.06(s，2H)4.23(d，J=5.37Hz，2H)6.75-6.81(m，1H)6.94-7.06(m，2H)7.13(s，1H)7.29(dd，J=8.66，1.46Hz，1H)7.45(d，J=8.54Hz，1H)7.50(s，1H)7.96(d，J=8.05Hz，1H)8.00(br.s.，1H)10.14(br.s.，1H)10.54(s，1H)12.77(br.s.，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2-methoxyethyl) (methyl) amino]Methyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (2-methoxyethyl) (methyl) amino]Methyl } -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.116
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.45(m，2H)1.95(d，J=11.83Hz，2H)2.22(s，3H)2.52-2.57(m，2H)3.26(s，3H)3.43-3.53(m，6H)3.64(dd，J=6.95，2.93Hz，1H)3.80 -3.88(m，2H)4.05(s，2H)6.58(d，J=7.93Hz，1H)6.79(s，1H)6.95-7.06(m，3H)7.27(dd，J=8.66，1.46Hz，1H)7.43(d，J=8.54Hz，1H)7.52(s，1H)7.86(d，J=8.05Hz，1H)7.96(d，J=7.56Hz，1H)10.39(s，1H)12.71(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (pyrrolidin-1-ylmethyl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (pyrrolidin-1-ylmethyl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.117
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.45(m，2H)1.67-1.78(m，4H)1.90-1.98(m，2H)2.47(br.s.，2H)3.44-3.54(m，2H)3.56(br.s.，4H)3.59-3.71(m，1H)3.83(dt，J=11.65，3.69Hz，2H)4.05(s，2H)6.59(d，J=8.66Hz，1H)6.77(s，1H)6.92-7.07(m，3H)7.27(dd，J=8.66，1.59Hz，1H)7.42(d，J=0.49Hz，1H)7.52(s，1H)7.85(d，J=8.17Hz，1H)7.95(d，J=7.80Hz，1H)10.39(s，1H)12.71(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (morpholin-4-ylmethyl) -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (morpholin-4-ylmethyl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.118
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.44(m，2H)1.88-2.01(m，2H)2.39(br.s.，4H)3.45-3.46(m，2H)3.46-3.54(m，2H)3.61(br.s.，4H)3.65(d，1H)3.84(d，J=12.32Hz，2H)4.05(s，2H)6.60(d，J=8.41Hz，1H)6.79(s，1H)6.89-7.09(m，3H)7.28(dd，J=8.72，1.16Hz，1H)7.43(d，J=8.78Hz，1H)7.51(s，1H)7.87(d，J=8.05Hz，1H)7.95(d，J=7.80Hz，1H)10.40(s，1H)12.71(s，1H)
4- (azetidin-1-ylmethyl)Yl) -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (azetidin-1-ylmethyl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.119
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.43(m，2H)1.90-1.98(m，2H)1.98-2.06(m，2H)3.15(t，J=6.95Hz，4H)3.46-3.54(m，4H)3.61-3.70(m，1H)3.79-3.88(m，2H)4.05(s，2H)6.53(dd，J=8.11，1.16Hz，1H)6.72(s，1H)6.94-7.05(m，3H)7.27(dd，J=8.60，1.52Hz，1H)7.43(d，J=8.66Hz，1H)7.51(s，1H)7.83(d，J=8.17Hz，1H)7.94(d，J=7.80Hz，1H)10.38(s，1H)12.70(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl } -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.127
1H-NMR(400MHz)，(ppm，DMSO-d6)：ppm3.44-3.56(m，2H)3.61-3.75(m，1H)3.78-3.88(m，2H)4.06(s，2H)4.26(br.s.，1H)6.66(s，1H)6.83(s，1H)6.95-7.06(m，3H)7.28(dd，J=8.66，1.46Hz，1H)7.44(d，J=8.78Hz，1H)7.53(s，1H)7.93(d，J=8.17Hz，1H)7.97(br.s.，1H)10.55(s，1H)12.75(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl } -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.128
1H-NMR(400MHz)，(ppm，DMSO-d6)：ppm3.44-3.56(m，2H)3.61-3.75(m，1H)3.78-3.88(m，2H)4.06(s，2H)4.26(br.s.，1H)6.66(s，1H)6.83(s，1H)6.95-7.06(m，3H)7.28(dd，J=8.66，1.46Hz，1H)7.44(d，J=8.78Hz，1H)7.53(s，1H)7.93(d，J=8.17Hz，1H)7.97(br.s.，1H)10.55(s，1H)12.75(s，1H)
N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]-N4-methyl-2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- ({ N- [2- (dimethylamino) ethyl]-N-methyl } carbonyl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.129
1H-NMR (400MHz), (ppm, DMSO-d 6): rotamer mixture 1.31-1.44(m, 2H)1.87-1.97(m, 2H)3.45-3.52(m, 2H)3.62-3.72(m, 1H)3.79-3.88(m, 2H)4.06(s, 2H)6.56(d, J =7.68Hz, 1H)6.76(br.s., 1H)6.95-7.05(m, 3H)7.28(dd, J =8.59, 1.52Hz, 1H)7.44(d, J =8.65Hz, 1H)7.54(s, 1H)7.91-7.99(m, 2H)10.56(s, 1H)12.75(s, 1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [4- (propan-2-yl) piperazin-1-yl]Carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [4- (propan-2-yl) piperazin-1-yl]Carbonyl } -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.130
1H-NMR(400MHz)，(ppm，DMSO-d6)：0.99(d，J=6.46Hz，6H)1.32-1.43(m，2H)1.89-1.97(m，2H)2.36-2.54(m，4H)2.66-2.75(m，1H)3.28-3.37(m，2H)3.49(td，J=11.18，2.13Hz，2H)3.61(br.s.，2H)3.65-3.74(m，1H)3.80-3.87 (m，2H)4.06(s，2H)6.58(dd，J=7.98，1.28Hz，1H)6.77(d，J=0.85Hz，1H)6.95-7.05(m，3H)7.28(dd，J=8.59，1.52Hz，1H)7.44(d，J=8.65Hz，1H)7.53(s，1H)7.91-7.95(m，1H)7.94-7.96(m，1H)10.56(s，1H)12.75(s，1H)
N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- [2- (dimethylamino) ethyl group]-2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- ({ N- [2- (dimethylamino) ethyl]} carbonyl) -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.131
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33-1.47(m，2H)1.93-2.00(m，2H)2.30(br.s.，6H)2.51-2.60(m，2H)3.37-3.44(m，2H)3.46-3.54(m，2H)3.73(d，1H)3.85(dt，J=11.61，3.76Hz，2H)4.06(s，2H)6.95-7.05(m，3H)7.07(dd，J=8.17，1.46Hz，1H)7.23(d，J=1.22Hz，1H)7.28(dd，J=8.65，1.58Hz，1H)7.44(d，J=8.53Hz，1H)7.54(s，1H)7.93(d，J=7.68Hz，1H)7.96(d，J=8.29Hz，1H)8.47(t，J=4.94Hz，1H)10.60(s，1H)12.76(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [ (4-methylpiperazin-1-yl) carbonyl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (4-methylpiperazin-1-yl) carbonyl]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.132
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.32-1.43(m，2H)1.93(d，J=11.70Hz，2H)2.22(s，3H)2.33(m，4H)3.45-3.53(m，2H)3.65-3.73(m，1H)3.80-3.86(m，2H)4.06(s，2H)6.57(dd，J=7.98，1.28Hz，1H)6.77(d，J=0.98Hz，1H)6.96-7.05(m，3H)7.28(dd，J=8.59，1.52Hz，1H)7.44(d，J=8.53Hz，1H)7.52(s，1H)7.92-7.95(m，1H)7.94-7.97(m，1H)10.56(s，1H)12.75(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [4- (dimethylamino) piperidin-1-yl]Carbonyl } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [4- (dimethylamino) piperidin-1-yl]Carbonyl } -2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.133
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.25-1.46(m，4H)1.73(m，1H)1.84(m，1H)1.93(d，J=11.46Hz，2H)2.20(s，6H)2.34(m，1H)2.82(m，1H)3.04(m，1H)3.42-3.55(m，2H)3.65-3.76(m，2H)3.78-3.88(m，2H)4.06(s，2H)4.43(m，1H)6.58(dd，J=8.05，1.34Hz，1H)6.78(d，J=0.85Hz，1H)6.94-7.06(m，3H)7.28(dd，J=8.65，1.58Hz，1H)7.44(d，J=8.53Hz，1H)7.53(s，1H)7.90-7.96(m，2H)10.55(s，1H) 12.74(s，1H)
N1- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-N4- (1-methylpiperidin-4-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzene-1, 4-dicarboxamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ N- (1-methylpiperidin-4-yl) -carbonyl]-2- (tetrahydro-pyran-4-ylamino) -phenyl]cpd.134
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.32-1.47(m，2H)1.56-1.70(m，2H)1.74-1.83(m，2H)1.92-1.99(m，2H)2.02(br.s.，2H)2.21(s，3H)2.82(d，J=13.17Hz，2H)3.46-3.54(m，2H)3.67-3.80(m，2H)3.79-3.88(m，2H)4.06(s，2H)6.96-7.05(m，3H)7.08(dd，J=8.17，1.46Hz，1H)7.21(d，J=1.22Hz，1H)7.28(dd，J=8.65，1.46Hz，1H)7.42-7.46(m，1H)7.54(s，1H)7.92-7.97(m，2H)8.26(d，J=7.80Hz，1H)10.60(s，1H)12.77(s，1H)。
Example 8
Preparation of tert-butyl 4- { [ (2S) -1-methylpyrrolidin-2-yl ] methoxy } -2-nitrobenzoate
Toluene (15ml), CsCO, was added to a round bottom three-necked flask under an argon atmosphere3(1.6g, 5mmol), phosphine ligand 2- (di-tert-butylphosphino) -1,1' -binaphthyl (331mg, 0.83mmol) and Pd (dba)2(380mg, 0.66 mmol). The mixture was degassed and bubbled with argon for 5 minutes. Tert-butyl 4-bromo-2-nitrobenzoate (1g, 3.31mmol) and (S) - (-) -1-methyl-2-pyrrolidinemethanol (0.78ml, 6.62mmol) were then added and the mixture was heated to 100 ℃ for 18 h. The reaction was cooled to room temperature, quenched with 30ml water and extracted twice with 25ml AcOEt. Collecting the organic phase, adding Na2SO4Drying and evaporation of the solvent gave a red oil which was chromatographed on the Biotage SP1 Automation System (90: 10DCM/MeOH isocratic elution) to give the pure title compound as a pale yellow oil (460mg, 1.36mmol, 41% yield)
ESI(+)MS：m/z337(MH+)。
The following compounds were obtained by operating in the same manner:
4- [ (1-Methylpiperidin-4-yl) oxy ] -2-nitrobenzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.48(9H，s)1.68(m，2H)1.95(m，2H)2.20(m，5H)2.61(m，2H)4.60(m，1H)7.30(dd，J=8.78，2.56Hz，1H)7.54(d，J=2.56Hz，1H)7.79(d，J=8.78Hz，1H)
4- [2- (dimethylamino) ethoxy ] -2-nitrobenzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.49(9H，s)2.22(s，6H)2.65(t，J=5.61Hz，2H)4.21(t，J=5.63Hz，2H)7.30(dd，J=8.78，2.56Hz，1H)7.52(d，J=2.56Hz，1H)7.80(d，J=8.78Hz，1H)
4- { [ (3S) -1-Methylpyrrolidin-3-yl ] oxy } -2-nitrobenzoic acid tert-butyl ester
ESI(+)MS：m/z323(MH+)。
Preparation of tert-butyl 2-amino-4- { [ (2S) -1-methylpyrrolidin-2-yl ] methoxy } benzoate
Coupling the nitro-derivative 4- { [ (2S) -1-methylpyrrolidin-2-yl]Methoxy } -2-nitrobenzoic acid tert-butyl ester (460mg, 1.37mmol) was dissolved in 20ml MeOH and 130mg of Pd/C5% and 700mg (6.3mmol) of HCOONH were added under argon4. The mixture was refluxed at 80 ℃ for 1h, then cooled to room temperature and filtered through a small pad of celite, washing with MeOH. The solvent was then distilled off, the residue was dissolved in 20ml DCM and the residue was taken up in 20ml NaHCO3(10%) washed twice. With Na2SO4The collected organic extracts were dried, filtered and evaporated to dryness to give a brown oil (400mg, 1.31mmol, 95% yield) which was used in the next step without any further purification.
ESI(+)MS：m/z307(MH+)。
The following compounds were obtained by operating in the same manner:
2-amino-4- [ (1-methylpiperidin-4-yl) oxy ] benzoic acid tert-butyl ester
ESI(+)MS：m/z307(MH+)。
2-amino-4- [2- (dimethylamino) ethoxy ] benzoic acid tert-butyl ester
1NMR(400MHz)，(ppm，DMSO-d6)：1.51(s，9H)2.21(s，6H)2.61(t，J=5.79Hz，2H)4.00(t，J=5.79Hz，2H)6.11(dd，J=8.96，2.50Hz，1H)6.25(d，J=2.56Hz，1H)6.60(s，2H)7.56(d，J=8.90Hz，1H)
2-amino-4- { [ (3S) -1-methylpyrrolidin-3-yl ] oxy } benzoic acid tert-butyl ester
ESI(+)MS：m/z293(MH+)。
Preparation of tert-butyl 4- { [ (2S) -1-methylpyrrolidin-2-yl ] methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzoate
2-amino-4- { [ (2S) -1-methylpyrrolidin-2-yl]Tert-butyl methoxy } benzoate (400mg, 1.3mmol) was dissolved in 20ml DCM. tetrahydro-4H-pyran-4-one (0.19ml, 2.05mmol), TFA (0.29ml, 3.69mmol) and Me were added4BH(OAc)3(540mg, 2.05 mmol). The resulting slurry was stirred at room temperature overnight and then with 15ml NaHCO310% was quenched and extracted twice with 20ml DCM. Then using Na2SO4The organic layer was dried, filtered and concentrated to give a yellow oil (448mg, 1.15mmol, 88%) which was used in the next step without any further purification.
ESI(+)MS：m/z391(MH+)。
The following compounds were obtained by operating in the same manner:
4- [ (1-Methylpiperidin-4-yl) oxy ] -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z391(MH+)。
4- [2- (dimethylamino) ethoxy ] -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z365(MH+)。
4- { [ (3S) -1-Methylpyrrolidin-3-yl ] oxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z377(MH+)。
Preparation of tert-butyl 4- { [ (2S) -1-methylpyrrolidin-2-yl ] methoxy } -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoate
Coupling 4- { [ (2S) -1-methylpyrrolidin-2-yl]Tert-butyl methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzoate (448mg, 1.15mmol) was dissolved in 20ml DCM. TEA (0.18ml,1.3mmol) and TFAA (0.27ml,1.7mmol) were added, the reaction mixture was stirred at room temperature for 2 hours and 15ml NaHCO was used3And (5) quenching by 10%. The resulting mixture was extracted twice with 20ml DCM and Na2SO4Dried, filtered and concentrated in vacuo. The crude product was chromatographed on silica gel (DCM/MeOH 95: 5) to give a yellow oil (481mg, 1mmol, 87%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.00(qd，J=12.25，4.82Hz，1H)1.47(s，9H)1.51-1.64(m，1H)1.98(d，J=12.68Hz，2H)3.83(ddd，J=31.46，11.34，4.02Hz，2H)4.51(tt，J=11.95，3.90Hz，1H)7.02(br.s.，1H)7.21(d，J=6.95Hz，1H)7.95(d，J=8.78Hz，1H)
The following compounds were obtained by operating in the same manner:
4- [ (1-Methylpiperidin-4-yl) oxy ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z487(MH+)。
4- [2- (dimethylamino) ethoxy ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z461(MH+)。
4- { [ (3S) -1-Methylpyrrolidin-3-yl ] oxy } -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z473(MH+)。
Preparation of 4- { [ (2S) -1-Methylpyrrolidin-2-yl ] methoxy } -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid trifluoroacetate salt
Coupling 4- { [ (2S) -1-methylpyrrolidine-2-yl]Methoxy } -2- [ tetrahydro-2H-pyran-4-yl- (trifluoroacetyl) amino group]Tert-butyl benzoate (480mg, 1mmol) was dissolved in 20ml DCM. Is added inAnhydrous HCl4M in alkane (2.5ml, 10 mmol). The reaction was stirred at room temperature for 5 days, then HPLC analysis showed the desired product to be formed, but with about 30% of the by-product being deacetylated. The solvent was removed in vacuo, the resulting yellow powder was suspended in 15ml of DCM and TFAA (0.28ml, 2mmol) was added. The solid dissolved immediately, the mixture was stirred for 2 hours, and HPLC analysis then showed complete disappearance of the by-product. The solvent was evaporated to dryness to give a dark yellow solid, which was used in the next synthesis step without any further purification.
ESI(+)MS：m/z431(MH+)。
The following compounds were obtained by operating in the same manner:
4- [ (1-Methylpiperidin-4-yl) oxy ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl trifluoroacetate
ESI(+)MS：m/z431(MH+)。
4- [2- (dimethylamino) ethoxy ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl trifluoroacetate
ESI(+)MS：m/z405(MH+)。
Tert-butyl 4- { [ (3S) -1-methylpyrrolidin-3-yl ] oxy } -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoate
ESI(+)MS：m/z417(MH+)。
Step i'
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2S) -1-methylpyrrolidin-2-yl]Methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, ar =4- { [ (2S) -1-methylpyrrolidin-2-yl]Methoxy } -2- (tetrahydro-2H-pyran-4-ylamino) -phenyl]Preparation of cpd.94
4- { [ (2S) -1-Methylpyrrolidin-2-yl ] in a Nitrogen atmosphere]Methoxy } -2- [ tetrahydro-2H-pyran-4-yl- (trifluoroacetyl) amino group]Benzoic acid trifluoroacetate salt (1mmol, 531mg) was dissolved in DCM and 2 drops of anhydrous DMF were added. Oxalyl chloride (0.17ml,2mmol) was added and the mixture was stirred at room temperature for 2 hours. The solvent was evaporated to give a yellow powder. The solid was redissolved in THF under argon and cooled at-20 ℃. DIPEA (0.56ml, 3.2mmol) was added. 5- (3, 5-difluorobenzyl) -1H-indazol-3-amine dissolved in 10mL dry THF is then added dropwise over 15'. The reaction mixture was kept at-20 ℃ for 6 hours, then the temperature was allowed to rise at room temperature overnight. With 15mL NaHCO35% the reaction was quenched and extracted twice with AcOEt (15 ml). The solvent was then evaporated and the residue was redissolved in 20ml MeOH. TEA (10mmol, 1.5ml) was added and the mixture was heated to 65 ℃ for 3 hr. The reaction was then cooled to room temperature and the solvent was removed to give the crude product, which was purified by flash chromatography on silica gel (AcOEt/MeOH/NH)3Aq.85: 15: 05) to give the title compound as a white powder (258mg, 0.45mmol, 45%).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.28-1.44(m，2H)1.94(d，J=12.07Hz，2H)3.44-3.56(m，3H)3.59-3.73(m，1H)3.77-3.87(m，2H)4.05(s，2H)6.23(dd，1H)6.28(d，J=2.19Hz，1H)6.92-7.06(m，3H)7.27(dd，J=8.66，1.59Hz，1H)7.42(d，J=8.90Hz，1H)7.50(s，1H)7.88(d，J=8.90Hz，1H)8.27(d，J=7.80Hz，1H)10.24(s，1H)12.68(s，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [ (1-methylpiperidin-4-yl) oxy]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A)，R1=R2=R3=H，R=35-difluorophenyl, Ar =4- [ (1-methylpiperidin-4-yl) oxy]-2- (tetrahydro-2H-pyran-4-ylamino) -phenyl]cpd.95
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.36(ddd，J=9.82，3.66，3.48Hz，2H)1.70(m，2H)1.85-2.00(m，4H)2.22-2.30(m，5H)2.64-2.79(m，2H)3.44-3.54(m，2H)3.58-3.72(m，1H)3.82(dt，J=11.65，3.69Hz，2H)4.05(s，2H)4.51(br.s.，1H)6.20-6.30(m，2H)6.94-7.07(m，3H)7.27(dd，J=8.60，1.52Hz，1H)7.42(d，J=8.54Hz，1H)7.50(d，J=2.32Hz，1H)7.88(d，J=9.51Hz，1H)8.22(d，J=7.68Hz，1H)10.24(s，1H)12.68(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [2- (dimethylamino) ethoxy]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [2- (dimethylamino) ethoxy]-2- (tetrahydro-2H-pyran-4-ylamino) -phenyl]cpd.96
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.44(m，2H)1.94(d，J=10.73Hz，2H)2.42(br.s.，6H)2.89(br.s.，2H)3.49(t，J=9.88Hz，2H)3.60-3.73(m，1H)3.78-3.88(m，2H)4.05(s，2H)4.19(t，J=5.24Hz，2H)6.25(dd，J=8.84，2.38Hz，1H)6.29(d，J=2.32Hz，1H)6.93-7.06(m，3H)7.27(dd，J=8.60，1.52Hz，1H)7.43(d，J=8.78Hz，1H)7.50(s，1H)7.90(d，J=8.78Hz，1H)8.27(d，J=7.44Hz，1H)10.26(s，1H)12.68(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (3S) -1-methylpyrrolidin-3-yl]Oxy } -2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (3S) -1-methylpyrrolidin-3-yl]Oxy } -2- (tetrahydro-2H-pyran-4-ylamino)) -phenyl radical]cpd.97
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.44(m，2H)1.75-1.86(m，1H)1.93(d，J=10.49Hz，2H)2.32(s，4H)2.44(br.s.，1H)2.65-2.78(m，2H)2.79-2.88(m，1H)3.44-3.54(m，2H)3.58-3.71(m，1H)3.82(d，J=11.58Hz，2H)4.05(s，2H)4.96-5.01(m，1H)6.15-6.19(m，1H)6.19-6.20(m，1H)6.94-7.06(m，3H)7.26(dd，J=8.66，1.59Hz，1H)7.42(d，J=8.54Hz，1H)7.50(d，J=1.59Hz，1H)7.87(d，J=8.90Hz，1H)8.23(d，J=7.68Hz，1H)10.24(s，1H)12.67(s，1H)。
Example 9
Step u
Preparation of N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] -2-fluoro-5-formyl-benzamide
2-fluoro-5-formyl-benzoic acid (368mg, 2.187mmol) in toluene (22mL) was treated with thionyl chloride (1.59mL, 21.87mmol) and stirred at reflux temperature for 4 h. The volatiles were evaporated, the residue was dissolved in toluene (4mL) and evaporated to dryness to give an off-white solid, which was dissolved in dry THF (5mL) and added dropwise to a solution of 5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-ylamine (843mg, 1.68mmol) and DIPEA (0.88mL, 5.04mmol) in THF (10mL) cooled to 4 ℃. The reaction was allowed to gradually reach room temperature. After one night, the volatiles were evaporated. The crude product was dissolved in DCM (150mL) and washed with NaHCO3Aqueous solution (100mL), then washed with water, then brine. After drying over sodium sulfate, evaporation and purification on silica gel (eluent: DCM), 868mg of the title compound are obtained as a white solid in 79% yield.
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.98(s，2H)6.34(d，J=8.66Hz，1H)6.89-7.09(m，3H)7.22-7.34(m，15H)7.53 -7.64(m，1H)7.66(s，1H)8.14(br.s.，1H)8.32(d，J=4.51Hz，1H)10.05(s，1H)11.08(br.s.，1H)
Step i'
Preparation of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -2-fluoro-5-formyl-benzamide
Is used inTreatment of 4N HCl in alkane (2.8mL) in dry IIN- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] in an alkane (25mL)]-2-fluoro-5-formyl-benzamide (740mg, 1.137 mmol). The reaction was stirred at room temperature for 2 days. The volatile components were evaporated to dryness and the residue was dissolved in Et2O (10mL), stirred for 1h, filtered by suction, Et2O (10mL) was washed and dried at 50 ℃ in vacuo to give 358mg of the title compound as a white solid in 77% yield.
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.08(s，2H)6.88-7.09(m，3H)7.22-7.31(m，1H)7.45(d，J=8.41Hz，1H)7.62(t，J=9.57Hz，1H)7.71(s，1H)8.12-8.19(m，1H)8.35(d，J=5.61Hz，1H)10.08(s，1H)10.92(s，1H)12.80(br.s.，1H)
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2-fluoro-5- (4-methyl-piperazin-1-ylmethyl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = 2-fluoro-5- (4-methyl-piperazin-1-ylmethyl) -phenyl]Preparation of cpd.120
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] in THF (4mL) was treated with N-methylpiperazine (0.039mL, 0.367mmol) followed by acetic acid (0.024mL, 0.422mmol) under a nitrogen atmosphere at room temperature]-2-fluoro-5-formyl-benzeneFormamide (150mg, 0.367 mmol). After 0.5 h, sodium triacetoxyborohydride was added and the reaction was stirred overnight. EtOAc (25mL) and water (25mL) were added with concentrated NH4OH adjusted the pH to 11. The organic layer was separated and the aqueous layer was extracted twice with EtOAc (2 × 10 mL). The combined organic extracts were dried over sodium sulfate, evaporated to dryness and purified over silica gel (eluent: DCM: 7N NH in MeOH)3= 96: 4) 177mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.18(br.s.，3H)2.30-2.46(m，8H)3.51(br.s.，2H)4.07(s，2H)6.92-7.00(m，2H)6.99-7.06(m，1H)7.28(s，1H)7.28-7.35(m，1H)7.44(d，J=8.53Hz，1H)7.47-7.54(m，1H)7.65(br.s.，1H)7.67(br.s.，1H)10.66(br.s.，1H)12.75(br.s.，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2-fluoro-5- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Methyl benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = 2-fluoro-5- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Methyl-phenyl]cpd.121
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.67(m，6H)1.92(m，1H)2.14(m，1H)2.63(m，2H)2.82(m，1H)3.23-3.37(m，2H)4.07(s，2H)4.17(d，J=14.26Hz，1H)6.93-6.99(m，2H)6.99-7.07(m，1H)7.25-7.28(m，1H)7.28-7.33(m，1H)7.44(d，J=8.65Hz，1H)7.50(br.s.，1H)7.66(br.s.，2H)10.64(br.s.，1H)12.75(br.s.，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2-fluoro-5- (morpholin-4-ylmethyl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = 2-fluoro-5- (morpholin-4-ylmethyl) -phenyl]cpd.122
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.40(br.s.，4H)3.52(br.s.，2H)3.60(br.s.，4H)4.07(s，2H)6.94-6.99(m， 2H)6.99-7.07(m，1H)7.27(d，J=8.65Hz，1H)7.33(d，J=8.53Hz，1H)7.44(d，J=8.53Hz，1H)7.53(br.s.，1H)7.67(br.s.，2H)10.67(br.s.，1H)12.75(br.s.，1H)。
Example 10
Preparation of N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] -4-fluoro-isophtalamic acid
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] -2-fluoro-5-formyl-benzamide (88mg, 0.135mmol) in t-butanol (1.8mL) was treated first with 2-methyl-2-butene (0.079mL, 1.082mmol), then with sodium chlorite (37mg, 0.405mmol) and aqueous sodium dihydrogen phosphate (0.8mL) dropwise at room temperature. The reaction was stirred overnight, then EtOAc (30mL) was added and washed with water (25 mL). The aqueous layer was extracted twice with EtOAc (2 × 10 mL). The combined organic layers were washed with brine and evaporated to dryness to give 106mg of the title compound, which was used directly in the following step without further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.98(s，2H)6.33(d，J=8.53Hz，1H)6.78(br.s.，1H)6.90-7.07(m，3H)7.20-7.35(m，15H)7.43-7.54(m，1H)7.65(br.s.，1H)8.13(br.s.，1H)8.29(d，J=3.66Hz，1H)11.01(s，1H)13.12(br.s.，1H)
Preparation of N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] -2-fluoro-5- ((S) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -benzamide
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl in DCM (1.4mL) was treated with 1-hydroxybenzotriazole (25mg, 0.181mmol), EDCI (35mg, 0.181mmol), and (S) - (+) -1- (2-pyrrolidinylmethyl) pyrrolidine (0.03mL, 0.1813mmol)]-4-fluoro-Isophthalein (93mg, 0.139 mmol). After 1h, the reaction was diluted with DCM (25mL) and NaHCO3Aqueous solution (5mL), water (5mL) and finally brine. After drying over sodium sulfate, the solvent is evaporated and purified over silica gel (eluent: DCM, 7N NH in MeOH)395: 5) 92mg of the title compound are obtained in 85% yield in two steps.
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.97(s，2H)4.26(br.s.，1H)6.33(d，J=8.41Hz，1H)6.86-7.09(m，4H)7.15-7.35 (m，15H)7.38-7.46(m，1H)7.63(s，1H)7.68(br.s.，1H)7.82(br.s.，1H)10.96(br.s.，1H)
Step i'
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2-fluoro-5- ((S) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = 2-fluoro-5- ((S) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -phenyl]cpd.123
N- [5- (3, 5-difluoro-benzyl) -1-trityl-1H-indazol-3-yl ] in DCM (1mL) was treated with TFA (0.17mL, 2.24mmol)]-2-fluoro-5- ((S) -2-pyrrolidin-1-ylmethyl-pyrrolidine-1-carbonyl) -benzamide (90mg, 0.112 mmol). After 2 hours at room temperature, DCM (25mL) was added and NaHCO was used3The organic phase was washed with aqueous solution, water and brine. Dried over sodium sulfate, evaporated and purified over silica gel (eluent: DCM/MeOH/7N NH in MeOH)3= 9: 1: 0.1) to yield 42mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.07(s，2H)4.29(br.s.，1H)6.93-6.99(m，2H)6.99-7.06(m，1H)7.27(dd，J=8.53，1.34Hz，1H)7.42(br.s.，1H)7.65-7.74(m，2H)7.85(br.s.，1H)10.80(br.s.，1H)12.77(br.s.，1H)。
Example 11
Step i'
Preparation of methyl 4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3-nitrobenzoate
4- (methoxycarbonyl) -2-nitrobenzoic acid (4.8g, 21.3mmol) and thionyl chloride (15.5 mL) were stirred in dry THF (130mL) at 70 ℃ for 2 h. The volatiles were evaporated and the residue was dissolved in dry pyridine (100mL) at 0 ℃. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (4.6mg, 17.76mmol) in dry pyridine (10mL) was added to the cooled reaction mixture. The temperature was allowed to reach room temperature overnight. With NaHCO3The reaction was stopped by saturated solution and extracted with ethyl acetate. With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM/EtOH/7N NH in MeOH)3=95/5/0.5) to give 5.4gr (65% yield) of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：3.97(s，3H)4.08(s，2H)6.89-7.00(m，2H)6.99-7.07(m，1H)7.29(dd，J=8.66，1.46Hz，1H)7.45(d，J=8.66Hz，1H)7.76(s，1H)8.01(d，J=7.93Hz，1H)8.40(dd，J=7.93，1.59Hz，1H)8.58(d，J=1.46Hz，1H)11.22(s，1H)12.81(s，1H)
The following compounds were obtained by operating in the same manner:
4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } benzoic acid methyl ester
ESI(+)MS：m/z422(MH+)。
Preparation of 4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3-nitrobenzoic acid
4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -3-nitrobenzoic acid methyl ester (5.4g, 11.6mmol) was dissolved in THF (78mL) and water (52mL) and treated with LiOH hydrate (730mg) at room temperature for 24 h. THF was evaporated and treated with 5% KHSO4Treating the resulting aqueous solution (100mL)An aqueous phase. The material thus precipitated was filtered off and dried in vacuo at 60 ℃ to give the title compound without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.08(s，2H)6.92-7.00(m，2H)7.00-7.07(m，1H)7.27(dd，J=8.59，1.40Hz，1H)7.44(d，J=8.65Hz，1H)7.76(s，1H)7.85(d，J=7.68Hz，1H)8.30(dd，J=7.74，1.28Hz，1H)8.50(d，J=1.22Hz，1H)11.08(s，1H)12.77(s，1H)
The following compounds were obtained by operating in the same manner:
4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } benzoic acid
ESI(+)MS：m/z408(MH+)。
Preparation of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazine-1-carbonyl) -2-nitro-benzamide
4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3-nitrobenzoic acid (500mg, 1.11mmol) in DMF (10mL) was treated with 1-hydroxybenzotriazole (195mg, 1.44mmol), EDCI (276mg, 1.44mmol) and 1-methylpiperazine (0.16mL, 1.44 mmol). The reaction was kept at room temperature overnight. The volatiles were removed by evaporation and the residue was added dropwise to ice water (25mL) with stirring. The resulting yellow solid was extracted with DCM (2 × 25 mL). The combined organic layers were dried over sodium sulfate and evaporated to give 590mg of the title compound, which was used in the next step without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.23(s，3H)2.34(m，2H)2.45(m，2H)3.39(m，2H)3.67(m，2H)4.08(s，2H)6.93-6.99(m，2H)6.99-7.07(m，1H)7.28(dd，J=8.59，1.40Hz，1H)7.45(d，J=8.53Hz，1H)7.74(s，1H)7.87-7.90(m，1H)7.90-7.93(m，1H)8.15(d，J=0.85Hz，1H)11.10(s，1H)12.78(s，1H)
The following compounds were obtained by operating in the same manner:
N-[5-(35-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (2R) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl group-phenyl group]cpd.124
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.06(s，2H)4.30(br.s.，1H)6.94-7.00(m，2H)6.99-7.06(m，1H)7.27(dd，J=8.66，1.59Hz，1H)7.44(d，J=8.54Hz，1H)7.61(d，2H)7.63(s，1H)8.11(d，J=8.29Hz，2H)10.81(s，1H)12.77(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl]Carbonyl group-phenyl group]cpd.125
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.06(s，2H)4.30(br.s.，1H)6.94-7.00(m，2H)6.99-7.06(m，1H)7.27(dd，J=8.60，1.52Hz，1H)7.44(d，J=8.90Hz，1H)7.59-7.65(m，3H)8.11(d，J=8.17Hz，2H)10.81(s，1H)12.77(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- { [4- (pyrrolidin-1-yl) piperidin-1-yl]Carbonyl benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- { [4- (pyrrolidin-1-yl) piperidin-1-yl]Carbonyl group-phenyl group]cpd.126
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.41(m，2H)1.70(m，4H)1.94(m，2H)2.33(m，1H)2.53(m，4H)3.06(m，2H)3.52(m，1H)4.06(s，2H)4.30m，1H)6.94-7.00(m，2H)7.00-7.05(m，1H)7.27(dd，J=8.66，1.59Hz，1H)7.42-7.46(m，1H)7.54(d，J=8.29Hz，2H)7.62(s，1H)8.12(d，J=8.17Hz，2H)10.81(s，1H)12.77(s，1H)。
Example 12
Preparation of 1-tert-butyl 4-methyl 2-nitro-terephthalate
Commercial 4-methyl 2-nitro-terephthalate (4.84g, 21.49mmol) in DCM (54mL) was treated with t-butanol (4.05mL, 42.99mmol), di-tert-butyl dicarbonate (12.19g, 55.87g) and DMAP (0.79g, 6.45 mmol). After 4 days at room temperature, the reaction was diluted with DCM (100mL) and 1N HCl (100mL), NaHCO3Aqueous solution and finally washed with water. After drying over sodium sulfate and evaporation of the volatiles, the title compound was obtained as a light brown oil in excess of quantitative yield (6.51 g). The crude product was used in the next step without further purification.
ESI(+)MS：m/z282(MH+)。
Preparation of 1-tert-butyl 2-nitro-terephthalate
1-tert-butyl 4-methyl 2-nitro-terephthalate (21.49mmol) was dissolved in THF (143mL) and treated with lithium hydroxide monohydrate (1.35g, 32.24mmol) in water (97 mL). The reaction was stirred at room temperature for 2 hours, then partially evaporated, cooled with an ice/water bath and treated by dropwise addition of 1N HCl (35 mL). A solid precipitate was produced. The mixture was then extracted with DCM (150mL and 2X50 mL). The aqueous phase was further treated with 1N HCl (10mL) and extracted with DCM (2X50 mL). The combined organic layers were then washed with water and finally brine. After drying over sodium sulfate and evaporation, 5.34g of the title compound are obtained as a pale red solid in a total yield of 93%.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.53(s，9H)7.93(d，J=7.92Hz，1H)8.31(dd，J=7.92，1.58Hz，1H)8.42(d，J=1.34Hz，1H)13.78(s，1H)
Preparation of 2-nitro-4- (piperidine-1-carbonyl) -benzoic acid tert-butyl ester
1-tert-butyl 2-nitro-terephthalate (500mg, 1.88mmol) in DCM (18mL) was treated with 1-hydroxybenzotriazole (0.39g, 2.43mmol), EDCI (0.47g, 2.43mmol) and piperidine (0.24mL, 2.43 mmol). After 3h, the reaction was diluted with DCM (50mL) and NaHCO3Aqueous solution (30mL), water (30mL) and finally brine. After drying over sodium sulfate and evaporation of the solvent, the title compound was obtained in quantitative yield as a colorless oil. The crude product was used without any further purification in the following reaction.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.45(br.s.，6H)1.51(s，9H)3.19-3.27(m，2H)3.59(br.s.，2H)7.76-7.81(m，1H)7.85-7.89(m，1H)8.01(d，J=1.22Hz，1H)
The following compounds were obtained by working in the same manner as described above:
4- [ (2-methoxyethyl) (methyl) carbamoyl ] -2-nitrobenzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6): rotamer mixture 1.52(s, 9H)7.77-7.83(m, 1H)7.84-7.91(m, 1H)8.03(d, J =0.61Hz, 1H)
2-Nitro-4- (pyrrolidin-1-ylcarbonyl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.52(s，9H)1.78-1.94(m，4H)3.37-3.43(m，2H)3.49(t，J=6.70Hz，2H)7.84-7.90(m，1H)7.91-7.96(m，1H)8.12(d，J=1.34Hz，1H)
4- (azetidin-1-ylcarbonyl) -2-nitrobenzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.52(s，9H)2.29(dt，J=15.51，7.79Hz，2H)4.06-4.12(m，2H)4.31-4.38(m，2H)7.88(d，J=7.92Hz，1H)8.01(dd，J=7.92，1.58Hz，1H)8.16(d，J=1.34Hz，1H)
4- (Morpholin-4-ylcarbonyl) -2-nitrobenzoic acid tert-butyl ester
ESI(+)MS：m/z337(MH+)。
Preparation of 2-nitro-4-piperidin-1-ylmethyl-benzoic acid hydrochloride
2-Nitro-4- (piperidine-1-carbonyl) -benzoic acid tert-butyl ester (1.87mmol) was dissolved in dry THF at room temperature under nitrogen with stirring and added dropwise to a 1.0M solution of 3.7mL borane tetrahydrofuran complex. The reaction was then refluxed for 6 hours, cooled to room temperature and cautiously treated with 2N HCl (10 mL). After stirring for 15 minutes, the solid K is added portionwise2CO3(1.75 g). The mixture was extracted with EtOAc (3 × 25 mL). The combined organic layers were dried over sodium sulfate and evaporated to give an oil, which was analyzed by HPLC-MS to show that the resulting 4: 6, and mixing the mixture. The mixture was dissolved in DCM (1mL) and used in ErbBTreatment with 4N HCl in alkane (7 mL). After 4 days at room temperature, an off-white colour formed, filtered and filteredAlkane (5mL) was washed and dried at 50 ℃ in vacuo. 0.40g of the title compound is obtained in a total yield of 70%.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.37-1.80(m，5H)2.90(br.s.，4H)4.42(s，2H)7.92-7.99(m，2H)8.24(d，J=0.85 Hz，1H)9.99(br.s.，1H)
4- { [ (2-methoxyethyl) (methyl) amino ] methyl } -2-nitrobenzoate
ESI(+)MS：m/z269(MH+)。
2-Nitro-4- (pyrrolidin-1-ylmethyl) benzoate
ESI(+)MS：m/z251(MH+)。
4- (morpholin-4-ylmethyl) -2-nitrobenzoate
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.39(t，J=4.51Hz，4H)3.59(t，J=4.63Hz，4H)3.62(s，2H)7.72(dd，J=7.87，1.28Hz，1H)7.82(d，J=7.80Hz，1H)7.87(d，J=0.98Hz，1H)
4- (azetidin-1-ylmethyl) -2-nitrobenzoate
ESI(+)MS：m/z237(MH+)。
Step i'
Preparation of N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-nitro-4- (piperidin-1-ylmethyl) benzamide
2-Nitro-4-piperidin-1-ylmethyl-benzoic acid hydrochloride (440mg, 1.46mmol) was treated with thionyl chloride (5mL) and refluxed for 1 hour. Excess reagent was removed by evaporation, followed by evaporation from toluene (2 × 5 mL). The solid was further dried in vacuo. The acid chloride was treated with dry pyridine (7mL), cooled to 4 ℃ and 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (315mg, 1.22mmol) in dry pyridine (3mL) was added under nitrogen with stirring. After stirring for several hours, the reaction was kept at 0 ℃ overnight. EtOAc (50mL) and water (50mL) were added with concentrated NH4OH adjusted the pH to 9. The organic layer was separated, dried over sodium sulfate, evaporated to dryness and purified on silica gel (DCM: MeOH = 95: 5) to give 266mg of the title compound in 43% yield.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.54(br.s.，6H)2.39(br.s.，4H)3.61(s，2H)4.07(s，2H)6.92-6.99(m，2H)6.99-7.06(m，1H)7.26-7.29(m，1H)7.44(d，J=8.53Hz，1H)7.73(s，1H)7.79(s，2H)8.04(s，1H)11.01(s，1H)12.75(s，1H)
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2-methoxyethyl) (methyl) amino ] methyl } -2-nitrobenzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.24(s，3H)2.60(t，J=5.67Hz，2H)3.26(s，3H)3.50(t，J=5.73Hz，2H)3.70(s，2H)4.07(s，2H)6.96(d，J=6.70Hz，2H)6.99-7.07(m，1H)7.24-7.30(m，1H)7.44(d，J=8.53Hz，1H)7.73(s，1H)7.80(s，2H)8.07(s，1H)11.02(s，1H)12.75(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-nitro-4- (pyrrolidin-1-ylmethyl) benzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.75(br.s.，4H)2.46-2.56(m，4H)3.77(br.s.，2H)4.07(s，2H)6.96(d，J=6.58Hz，2H)6.99-7.06(m，1H)7.25-7.30(m，1H)7.44(d，J=8.54Hz，1H)7.73(s，1H)7.80(s，2H)8.05(s，1H)11.02(s，1H)12.75(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (morpholin-4-ylmethyl) -2-nitrobenzamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.40-2.46(m，4H)3.60-3.65(m，4H)3.66(s，2H)4.07(s，2H)6.90-6.99(m，2H)6.99-7.07(m，1H)7.24-7.29(m，1H)7.44(d，J=8.54Hz，1H)7.73(s，1H)7.81(s，2H)8.07(s，1H)11.02(s，1H)12.75(s，1H)
4- (azetidin-1-ylmethyl) -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2-nitrobenzamide
ESI(+)MS：m/z478(MH+)。
Transformation 4
Preparation of 2-amino-N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4-piperidin-1-ylmethyl-benzamide
Reacting N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl]-2-Nitro-4-piperidin-1-ylmethyl-benzamide (255mg, 0.505mmol) was suspended in DCM (7mL) and treated with nBu4NCl (95mg, 0.343 mmol). Na in water (3.4mL) was added dropwise with stirring2S2O4(659mg, 3.029 mmol). After 2 hours, the medicine is turned onVolatiles were removed by evaporation, the solid was filtered from the aqueous phase and dried in vacuo. Is used inThe solid was treated with 4N HCl in an alkane (12mL) and then the solvent was removed by evaporation. Dissolve the solid in DCM (100mL) with K2CO3The aqueous solution was then washed with brine. After drying over sodium sulfate and removal of the solvent, 248mg of the title compound are obtained in more than quantitative yield. The crude product was used in the next step without further purification.
ESI(+)MS：m/z476(MH+)。
The following compounds were obtained by working in the same manner as described above:
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- { [ (2-methoxyethyl) (methyl) amino ] methyl } -benzamide
ESI(+)MS：m/z480(MH+)。
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (pyrrolidin-1-ylmethyl) benzamide
ESI(+)MS：m/z462(MH+)。
2-amino-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- (morpholin-4-ylmethyl) -benzamide
ESI(+)MS：m/z478(MH+)。
2-amino-4- (azetidin-1-ylmethyl) -N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -benzamide
ESI(+)MS：m/z448(MH+)。
Example 13
Preparation of tert-butyl 4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) amino ] benzoate
Tert-butyl 2-amino-4- (4-methylpiperazin-1-yl) benzoate (1.5g, 5.15mmol) was dissolved in dry di-n-butyl ester under nitrogen atmosphereAlkane (25 mL). N-methylpiperidinone (0.72g, 6.18mmol, 1.2eq) was added followed by trifluoroacetic acid (1.03mL, 13.39mmol, 2.6eq) and sodium triacetoxyborohydride (1.72g, 7.73mmol, 1.5 eq). The mixture was stirred at room temperature for 26 hours. During this time, additional portions of N-methylpiperidinone (0.5mL, 0.75eq) and sodium triacetoxyborohydride (1.72g, 7.73mmol, 1.5eq) were added. Then NaHCO is added3Saturated aqueous solution, the reaction mixture was concentrated under reduced pressure. 10% ammonium hydroxide was added to pH10 and the aqueous phase was extracted with dichloromethane. The organic phase was washed with brine and Na2SO4Dried and evaporated under reduced pressure. Purification by silica gel chromatography (DCM/MeOH/NH in MeOH)37% = 90: 8: 2) this gave 1.025g of the title compound as an off-white solid (51% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.36-1.47(m，2H)1.50(s，9H)1.88-1.98(m，2H)2.09-2.16(m，2H)2.18(s，3H)2.21(s，3H)2.38-2.44(m，4H)2.59-2.68(m，2H)3.20-3.26(m，4H)3.37-3.50(m，1H)6.01(d，J=1.95Hz，1H)6.18(dd，J=9.08，2.26Hz，1H)7.56(d，J=9.02Hz，1H)7.68(d，J=7.56Hz，1H)
The following compounds were obtained by operating in the same manner:
4- { [2- (tert-Butoxycarbonyl) -5- (4-methylpiperazin-1-yl) phenyl ] amino } piperidine-1-carboxylic acid ethyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.19(t，J=7.50Hz，3H)1.24-1.34(m，2H)1.50(s，9H)1.89-2.00(m，2H)2.22(s，3H)2.39-2.45(m，4H)3.03-3.16(m，2H)3.20-3.29(m，4H)3.66-3.76(m，1H)3.80-3.90(m，2H)4.05(q，J=7.07Hz，2H)6.07(d，J=2.07Hz，1H)6.20(dd，J=9.15，2.19Hz，1H)7.57(d，J=9.02Hz，1H)7.70(d，J=7.93Hz，1H)
Preparation of tert-butyl 4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) (trifluoroacetyl) amino ] benzoate
4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) amino]Tert-butyl benzoate (1.02g, 2.625mmol) was dissolved in dry dichloromethane (10mL) and the solution was cooled to 0 ℃. Triethylamine (0.548mL, 3.938mmol, 1.5eq) was added followed by trifluoroacetic anhydride (0.445mL, 3.15mmol, 1.2eq) and the mixture was stirred at 0 ℃ for 2 h. It was then diluted with dichloromethane and washed twice with water. The aqueous phase was back-extracted with dichloromethane. With Na2SO4The combined organic phases were dried and concentrated under reduced pressure to give 1.18g of crude product (93% yield), which was used in the next step without further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：0.93-1.07(m，2H)1.45(s，9H)1.48-1.64(m，2H)1.85-2.05(m，2H)2.11(s，3H)2.23(s，3H)2.41-2.47(m，4H)2.66-2.87(m，2H)3.27-3.35(m，4H)4.10-4.26(m，1H)6.78(d，J=2.44Hz，1H)7.05(dd，J=9.02，2.56Hz，1H)7.81(d，J=9.02Hz，1H)
The following compounds were obtained by operating in the same manner:
4- { [2- (tert-butoxycarbonyl) -5- (4-methylpiperazin-1-yl) phenyl ] (trifluoroacetyl) amino } piperidine-1-carboxylic acid ethyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：0.77-0.93(m，1H)1.13(t，J=7.07Hz，3H)1.34-1.44(m，1H)1.46(s，9H)1.56-1.63(m，1H)2.01-2.10(m，1H)2.22(s，3H)2.40-2.44(m，4H)2.78-2.97(m，2H)3.27-3.36(m，4H)3.91-4.06(m，2H)3.94-4.01(m，2H)4.37-4.47(m，1H)6.78(d，J=2.44Hz，1H)7.04(dd，J=9.02，2.56Hz，1H)7.81(d，J=9.02Hz，1H)
Preparation of 4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) (trifluoroacetyl) amino ] benzoic acid dihydrochloride
4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) (trifluoroacetyl) amino group was reacted under a nitrogen atmosphere]Tert-butyl benzoate (1.18g,2.435mmol) was dissolved in dry dichloromethane (3 mL). Then 4M HCl is added dropwise inSolution in alkane (9.1mL, 36.4mmol, 15eq) and the mixture was stirred for 1.5 h. A sticky solid was formed. More than 5 equivalents of HCl are added and the mixture is stirred for more than 2 hours. The solid was filtered, washed with DCM (10mL) and diethyl ether (10mL), and dried in vacuo at 60 ℃ for 2 h. 1.06g of the title compound are obtained as a light brown powder (87% yield).
ESI(+)MS：m/z429(MH+)。
The following compounds were obtained by operating in the same manner:
2- { [1- (ethoxycarbonyl) piperidin-4-yl ] (trifluoroacetyl) amino } -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：0.83-0.98(m，1H)1.13(t，J=7.01Hz，3H)1.34-1.47(m，1H)1.63(d，J=10.85Hz， 1H)2.04(d，J=13.66Hz，1H)2.84(s，3H)2.88(m，2H)3.16(m，4H)3.52(m，2H)3.94-4.02(m，2H)4.05(m，4H)4.34-4.48(m，1H)6.96(d，J=2.32Hz，1H)7.11(dd，J=8.90，2.56Hz，1H)7.91(d，J=8.90Hz，1H)10.26(br.s.，1H)12.79(br.s.，1H)
Step i'
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) amino]Benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl)]Amino-phenyl]Preparation of cpd.13
4- (4-methylpiperazin-1-yl) -2- [ (1-methylpiperidin-4-yl) (trifluoroacetyl) amino group was reacted under a nitrogen atmosphere]Benzoic acid dihydrochloride (251mg, 0.501mmol, 1.3eq) was mixedSuspended in dry THF (4 mL). Thionyl chloride (0.365mL, 1.0mmol, 2.6eq) was added and the mixture was stirred at 70 ℃ for 1.5 h. The mixture was then evaporated to dryness, dissolved in toluene and evaporated to dryness, and then kept at room temperature under high vacuum for 2 hours. The acid chloride was then suspended in dry pyridine (2mL) and cooled to 0 ℃. A solution of 5- (3, 5-difluorobenzyl) -1H-indazol-3-amine (100mg, 0.386mmol, 1eq) in dry pyridine (1.2mL) was added dropwise, and the mixture was stirred at 0 ℃ for 2 hours and then held at 4 ℃ overnight. Then diluted with water and ethyl acetate. The aqueous phase was basified to pH10 with 30% ammonium hydroxide and extracted with ethyl acetate. With Na2SO4The combined organic layers were dried and evaporated to dryness to give 290mg of crude trifluoroacetamide. The crude product was dissolved in methanol (7mL), triethylamine (1.3mL, 9.34mmol, 24eq) was added and the solution was refluxed for 1.5 h. The reaction mixture was evaporated to dryness and purified by silica gel chromatography (DCM/MeOH/NH in MeOH)37% = 83: 17: 1). The product was then stirred into a slurry in ether (1mL) at room temperature for 30 minutes, then filtered and dried at 45 ℃ under high vacuum for 3 hours. 153mg of the title compound are obtained as a pale yellow powder (69% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33-1.50(m，2H)1.92(dd，J=9.51，4.02Hz，2H)2.18(br.s.，3H)2.21(br.s.，2H)2.23(s，3H)2.44(t，J=4.60Hz，4H)2.61(br.s.，2H)3.25(t，J=4.90Hz，4H)3.41-3.52(m，1H)4.04(s，2H)6.08(d，J=1.95Hz，1H)6.22(dd，J=8.96，2.13Hz，1H)6.98(m，3H)7.24(dd，J=8.65，1.46Hz，1H)7.40(d，J=8.53Hz，1H)7.49(s，1H)7.78(d，J=9.02Hz，1H)8.26(d，J=7.44Hz，1H)10.06(s，1H)12.62(s，1H)
The following compounds were obtained by operating in the same manner:
4- { [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl]Amino } piperidine-1-carboxylic acid ethyl ester [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methylpiperazin-1-yl) -2- { (1- (ethoxycarbonyl) piperidin-4-yl]Amino } -phenyl radical]cpd.138
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.17(t，J=7.07Hz，3H)1.21-1.34(m，2H)1.87-1.98(m，2H)2.26(br.s.，3H)2.45-2.49(m，4H)3.07-3.21(m，2H)3.25-3.35(m，4H)3.64-3.73(m，1H)3.76(ddd，J=13.57，4.18，3.96Hz，2H)4.02(q，J=7.03Hz，2H)4.04(s，2H)6.15(d，J=2.10Hz，1H)6.25(dd，J=9.11，2.10Hz，1H)6.92-7.05(m，3H)7.25(dd，J=8.57，1.52Hz，1H)7.41(d，J=8.57Hz，1H)7.47(s，1H)7.80(d，J=9.11Hz，1H)8.31(d，J=7.93Hz，1H)10.09(s，1H)12.63(s， 1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-methylpiperazin-1-yl) -2- (piperidin-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methylpiperazin-1-yl) -2- [ (piperidin-4-yl) amino]-phenyl radical]Preparation of cpd.139
4- { [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl in a screw-cap pirex tube]Carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl]Amino } piperidine-1-carboxylic acid ethyl ester (198mg, 0.313mmol) was dissolved in 62% aqueous HBr (4mL) and stirred at 70 ℃ for 1 h. The mixture was then diluted with water and 30% ammonium hydroxide and extracted with ethyl acetate. With Na2SO4The organic phase was dried and concentrated to dryness. Purification by silica gel chromatography (DCM/MeOH/NH in MeOH)37% = 80: 10: 10) 127mg of pure product are obtained (72% yield). The product was stirred with ethyl acetate as a slurry, filtered, washed with n-hexane and dried at 45 ℃ under high vacuum for 3 hours to give 88mg of the title compound as a white solid.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.16-1.31(m，J=12.50，10.20，10.20，3.66Hz，2H)1.89(dq，J=12.50，3.40Hz，2H)2.22(s，3H)2.43(t，J=4.76Hz，4H)2.63(ddd，J=12.59，10.27，2.62Hz，2H)2.92(dt，J=12.53，3.92Hz，2H)3.25(t，J=4.63Hz，4H)3.46-3.57(m，1H)4.04(s，2H)6.09(d，J=2.07Hz，1H)6.22(dd，J=9.02，2.07Hz，1H)6.93-7.04(m，3H)7.24(dd，J=8.66，1.59Hz，1H)7.40(d，J=8.66Hz，1H)7.48(br.s.，1H)7.78(d，J=9.02Hz，1H)8.24(d，J=7.80Hz，1H)10.04(s，1H)12.62(s，1H)。
Example 14
Preparation of 1- [1- (tert-butoxycarbonyl) piperidin-4-yl ] -1H-pyrazole-4-carboxylic acid
A mixture of ethyl 1H-pyrazole-4-carboxylate (700mg, 5mmol) and NaH60% (6mmol) was stirred in dry DMF (15mL) under nitrogen at 0 deg.C for 1H. 4- [ (methylsulfonyl) oxy) dissolved in 4mL dry DMF was added]Tert-butyl piperidine-1-carboxylate (1.53g, 5.5mmol), and the resulting solution was heated at 100 ℃ overnight. The reaction mixture was quenched with water and extracted with ethyl acetate (x 3). With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness. The residue was dissolved in MeOH (20mL) and water (5mL) and KOH (1.12g, 20mmol) was added. The resulting solution was stirred at room temperature for 24 hours, and then the solvent was removed under reduced pressure. The residue was dissolved in AcOEt and KHSO45% solution. The aqueous phase was extracted several times with EtOAc. With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness to yield 600mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.42(s，9H)1.73-1.87(m，2H)1.96-2.03(m，2H)2.82-2.99(m，2H)4.04(d，J=12.93Hz，2H)4.34-4.47(m，1H)7.81(s，1H)8.29(s，1H)12.26(br.s.，1H)
Step i'
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-1- (piperidin-4-yl) -1H-pyrazole-4-carboxamide hydrochloride [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar = (piperidin-4-yl) -1H-pyranylAzole]Preparation of cpd.102
1- [1- (tert-butoxycarbonyl) piperidin-4-yl]-1H-pyrazole-4-carboxylic acid (134mg, 0.45mmol) and oxalyl chloride (0.6mmol) in dry DCM (5mL) and stirred at room temperature overnight. The volatiles were evaporated and the residue was dissolved in dry pyridine (5mL) at 0 ℃. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (100mg, 0.38mmol) in dry pyridine (2mL) was added to the cooled reaction mixture. After 1 hour, NaHCO was used3The saturated solution quenched the reaction and was extracted with ethyl acetate. With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM/EtOH/NH in MeOH)35N =1000/50/1) to yield 87mg of Boc-protected derivative, which was dissolved in 2mL of twoAlkane and 0.4mL in twoTreatment with 4M HCl in alkane. The volatiles were evaporated to give 65mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.10-2.23(m，2H)2.22-2.31(m，2H)3.03-3.19(m，2H)3.32-3.49(m，2H)4.05(s，2H)4.54-4.63(m，1H)6.92-6.98(m，2H)6.98-7.05(m，1H)7.25(dd，J=8.59，1.65Hz，1H)7.40-7.44(m，1H)7.63(d，J=0.61Hz，1H)8.16(s，1H)8.49(s，1H)8.65-8.77(m，1H)8.82-8.96(m，1H)10.44(s，1H)12.71(br.s.，1H)
Example 15
Step i'
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (cis-4-hydroxycyclohexyl) amino]-4- (4-methylpiperOxazin-1-yl benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (cis-4-hydroxycyclohexyl) amino]-phenyl radical]Preparation of cpd.103
4- (4-methylpiperazin-1-yl) -2- [ { cis-4- [ (phenylcarbonyl) oxy group]Cyclohexyl } (trifluoroacetyl) amino]Benzoic acid hydrochloride (1.03g, 1.94mmol) and oxalyl chloride (3.88mmol) were stirred in dry DCM (20mL) and a few drops of dry DMF at 0 deg.C, and the temperature was allowed to reach room temperature over 2 hours. The volatiles were evaporated and the residue was dissolved in dry pyridine (25mL) at 0 ℃. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (387mg, 1.49mmol) in dry pyridine (6mL) was added to the cooled reaction mixture. The temperature was allowed to reach room temperature overnight. With NaHCO3The reaction was stopped by saturated solution and extracted with ethyl acetate. With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM/AcOEt/EtOH = 100/10/15). The derivative thus obtained was dissolved in MeOH (200mL) and water (20mL) and treated with LiOH hydrate (160mg, 3.8mmol) at 60 ℃ for 4 hours. MeOH was evaporated and the resulting aqueous phase was extracted with EtOAc. With Na2SO4The collected organic phases were dried, filtered and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM/EtOH/NH in MeOH)35N =100/10/2) to yield 233mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.41-1.70(m，8H)2.24(s，3H)2.45(br.s.，4H)3.22-3.29(m，4H)3.58(d，J=10.61Hz，2H)4.05(s，2H)4.43(d，J=3.78Hz，1H)6.09(d，J=1.95Hz，1H)6.22(dd，J=8.96，2.13Hz，1H)6.94-7.04(m，3H)7.25(dd，J=8.65，1.58Hz，1H)7.41(d，J=8.53Hz，1H)7.51(s，1H)7.79(d，J=9.14Hz，1H)8.39(d，J=7.68Hz，1H)10.04(s，1H)12.63(s，1H)
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (trans-4-hydroxycyclohexyl) amino]-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (trans-4-hydroxycyclohexyl) amino]-phenyl radical]cpd.104
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.10-1.22(m，2H)1.29-1.41(m，2H)1.78-1.83(m，2H)1.94-2.03(m，2H)2.24(s，3H)2.42-2.48(m，4H)3.23-3.28(m，4H)3.34-3.42(m，1H)3.43-3.52(m，1H)4.04(s，2H)4.53(d，J=4.14Hz，1H)6.09(d，J=2.07Hz，1H)6.21(dd，J=9.02，2.19Hz，1H)6.95-7.04(m，3H)7.25(dd，J=8.53，1.58Hz，1H)7.40(d，J=8.53Hz， 1H)7.48(s，1H)7.77(d，J=9.14Hz，1H)8.17(d，J=7.80Hz，1H)10.04(s，1H)12.61(s，1H)
Example 16
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- [ (2-hydroxyethyl) amino]-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (2-hydroxyethyl) amino]-phenyl radical]Preparation of cpd.105
2- [ (2- { [ tert-butyl (dimethyl) silyl group)]Oxy } ethyl) amino]-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-methylpiperazin-1-yl) benzamide (126mg, 0.2mmol) was dissolved in dry THF (3mL) and 1M TBAF in THF (0.24mL) was added at 0 ℃. The resulting solution was stirred at room temperature overnight. The reaction was stopped with water and extracted with ethyl acetate. With Na2SO4Drying and collectingThe organic phase of (a), was filtered and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM/EtOH/NH in MeOH)35N =85/15/1), yielding 83mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.34(br.s.，3H)2.51-2.65(m，4H)3.20(q，J=5.57Hz，2H)3.25-3.36(m，4H)3.60(q，J=5.53Hz，2H)4.05(s，2H)4.74(t，J=5.18Hz，1H)6.09(d，J=2.07Hz，1H)6.25(dd，J=8.90，2.19Hz，1H)6.94-6.99(m，2H)6.99-7.04(m，1H)7.23(dd，J=8.66，1.58Hz，1H)7.41(d，J=8.65Hz，1H)7.51(s，1H)7.79(d，J=9.02Hz，1H)8.22(t，J=5.18Hz，1H)10.06(s，1H)12.62(s，1H)
Example 17
2- [ (azetidin-3-ylmethyl) amino]-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (azetidin-3-ylmethyl) amino]-phenyl radical]Preparation of cpd.106
3- ({ [2- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -5- (4-methylpiperazin-1-yl) phenyl]Amino } methyl) azetidine-1-carboxylic acid tert-butyl ester (289mg, 0.45mmol) was dissolved in DCM (3mL) and TFA (0.7mL) was added. The resulting reaction solution was stirred at room temperature overnight. The mixture was diluted with DCM and diluted with 10% NH3And (4) water extraction. The organic phase was evaporated. Purification by reverse phase column chromatography gave 104mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.24(s，3H)2.42-2.47(m，4H)2.80-2.90(m，1H)3.26-3.38(m，4H)3.58(t，J=7.86Hz，2H)4.04(s，2H)6.08(d，J=2.32Hz，1H)6.25(dd，J=8.96，2.13Hz，1H)6.94-7.00(m，2H)6.98-7.04(m，1H)7.25(dd，J=8.65，1.58Hz，1H)7.39-7.43(m，1H)7.49(d，J=0.61Hz，1H)7.80(d，J=8.90Hz，1H)8.16(t，J=5.06Hz，1H)10.07(br.s.，1H)12.63(br.s.，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- { [ (1-methylazetidin-3-yl) methyl]Amino } -4- (4-methylpiperazin-1-yl) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methyl-piperazin-1-yl) -2- [ (1-methylazetidin-3-ylmethyl) amino]-phenyl radical]Preparation of cpd.107
To 2- [ (azetidin-3-ylmethyl) amino]-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]To a solution of-4- (4-methylpiperazin-1-yl) benzamide (100mg, 0.14mmol) in dichloromethane (2mL) was added 37wt.% formaldehyde in water (0.014mL, 0.168mmol), TEA (0.4mmol) and sodium triacetoxyborohydride (45mg, 0.21 mmol). The mixture was stirred at room temperature overnight, diluted with dichloromethane and diluted with NaHCO3Saturated aqueous solution, water and brine. The organic phase was dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on silica gel using dichloromethane/methanol/NH in MeOH35N (100: 10: 1) as eluent, 5mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.26(s，3H)2.47(br.s.，4H)2.62(s，3H)2.84-2.99(m，1H)3.27-3.34(m，4H)3.36-3.46(m，2H)3.52-3.62(m，2H)3.80-3.90(m，2H)4.04(s，2H)6.09(d，J=2.07Hz，1H)6.28(dd，J=9.02，2.07Hz，1H)6.93-6.99(m，2H)6.99-7.05(m，1H)7.25(dd，J=8.59，1.52Hz，1H)7.41(d，J=8.65Hz，1H)7.50(s，1H)7.81(d，J=9.14Hz，1H)8.25(t，J=5.49Hz，1H)10.12(s，1H)12.64(s，1H)。
Example 18
Preparation of 4-nitro-2- (tetrahydro-pyran-4-ylamino) -benzoic acid
4-Nitro-2- (tetrahydro-pyran-4-ylamino) -benzoic acid ethyl ester (11.2g, 38mmol) was dissolved in 200mL ethanol at 60 deg.C, followed by the addition of 2N NaOH (40mL, 80 mmol). The mixture was stirred at 60 ℃ for 4 hours, and then the solvent was removed under reduced pressure. The residue was dissolved in 200mL of water and the mixture was brought to acidic pH with 2N HCl (35 mL). The precipitated yellow solid was filtered, washed with copious amounts of water and dried in an oven at 40 ℃ to give the title compound (9.3 g).
1H-NMR(400MHz)，(ppm，DMSO-d6)：13.49(bs，1H)，8.17(bd，1H)，8.04(d，J=8.7Hz，1H)，7.54(d，J=2.2Hz，1H)，7.32(dd，J1=8.7HZ，J2=2.2Hz，1H)，3.90-3.78(m，3H)，3.54(m，2H)，1.98(m，2H)，1.46(m，2H)。
Preparation of 4-nitro-2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid
4-Nitro-2- (tetrahydro-pyran-4-ylamino) -benzoic acid (9.1g, 34.2mmol) was added in small portions to 30mL of trifluoroacetic anhydride at room temperature. The mixture was stirred at room temperature for 1 hour and then evaporated to dryness. The residue (brown oil) was treated with 200mL of water and stirred vigorously at room temperature for 3 hours. The white solid thus formed was filtered, washed with a large amount of water and dried in an oven at 40 ℃ to give the title compound (11.8 g).
1H-NMR(400MHz)，(ppm，DMSO-d6)：13.52(bs，1H)，8.45(dd，J1=8.5Hz，J2=2.3Hz，1H)，8.32(d，J=2.3Hz，1H)，8.26(d，J=8.5Hz，1H)，4.58(m，1H)，3.84(m，2H)，3.45-3.2(m，2H)，1.98(m，1H)，1.59(m，1H)，1.49(m，1H)，1.14(m，1H)。
Step i'
Preparation of N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4-nitro-2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzamide
Reacting 4-nitro-2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino]Benzoic acid (3.62g, 10 m)mol) and oxalyl chloride (3.8mL, 30mmol) in dry DCM (120mL) and a few drops of dry DMF, stirring at room temperature for 2 hours. The volatiles were evaporated and the residue was dissolved in dry pyridine (50mL) at 0 ℃. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (2g, 7.72mmol) in dry pyridine (20mL) was added to the cooled reaction mixture under a nitrogen atmosphere. The resulting mixture was allowed to react at room temperature overnight, and then the solvent was removed under reduced pressure. The residue was dissolved in EtOAc and taken up with NaHCO3Saturated aqueous solution, water and brine. The organic phase was dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on silica gel using AcOEt/hexanes 7: 3 as eluent, 3.9g of the title compound are obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.38-1.57(m，2H)1.65-1.74(m，1H)1.91-1.98(m，1H)3.25-3.44(m，2H)3.70-3.78(m，1H)3.87(dd，J=11.92，4.09Hz，1H)4.04(s，2H)4.47-4.58(m，1H)6.98(d，J=1.34Hz，2H)6.99-7.06(m，1H)7.31(dd，J=8.68，1.47Hz，1H)7.45(d，J=8.56Hz，1H)7.54(s，1H)8.20(d，J=8.56Hz，1H)8.36(d，J=2.32Hz，1H)8.51(dd， J=8.56，2.08Hz，1H)11.28(s，1H)12.85(s，1H)
Transformation 4
Preparation of N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4-amino-2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzamide
Reacting N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4-nitro-2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino]Benzamide (3.86g, 6.4mmol), cyclohexene (10mL), bisA mixture of an alkane (70mL) and 10% Pd/C (0.42g) was stirred at 100 ℃ for 4 hours. The reaction mixture was filtered through a pad of celite, washing well with THF and MeOH. After evaporation of the organic phase, the crude product was purified by silica gel chromatography (DCM/EtOH9/1) to yield 2.75g of the title compound (82% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29(qd，J=12.28，4.63Hz，1H)1.56(qd，J=12.19，4.51Hz，1H)1.62(ddd，J=12.93，3.47，2.01Hz，1H)1.84(ddd，J=12.47，3.93，2.01Hz，1H)3.33(m，2H)3.77(dd，J=11.58，4.39Hz，1H)3.88(dd，J=11.65，4.33Hz，1H)4.00(s，2H)4.43(tt，J=11.93，3.86Hz，1H)5.96(s，2H)6.50(d，J=2.32Hz，1H)6.68(dd，J=8.47，2.26Hz，1H)6.89-6.97(m，2H)7.01(tt，J=9.43，2.33Hz，1H)7.25(dd，1H)7.39(m，2H)7.68(d，J=8.54Hz，1H)10.33(s，1H)12.64(s，1H)
Transformation 6
Preparation of tert-butyl 3- { [ (4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] phenyl) amino ] methyl } azetidine-1-carboxylate
To N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4-amino-2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino]To a solution of benzamide (240mg, 0.42mmol) in dichloromethane (20mL) was added tert-butyl 3-formylazetidine-1-carboxylate (116mg, 0.63mmol), trifluoroacetic acid (0.32mL) and tetramethylammonium triacetoxyborohydride (165mg g, 0.63 mmol). The mixture was stirred at room temperature overnight, then diluted with dichloromethane and with NaHCO3The saturated solution was washed with brine, dried over sodium sulfate and evaporated to dryness.
ESI(+)MS：m/z743(MH+)。
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -4- [ (1-methylpiperidin-4-yl) amino ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzamide
ESI(+)MS：m/z671(MH+)。
Preparation of tert-butyl 3- ({ [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) phenyl ] amino } methyl) azetidine-1-carboxylate
Tert-butyl 3- { [ (4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] phenyl) amino ] methyl } azetidine-1-carboxylate (760mg, 1.02mmol) was dissolved in MeOH (12mL) and TEA (4mL) and stirred at room temperature overnight. The volatiles were evaporated and the residue was dissolved in DCM and washed with brine. The organic phase was dried over sodium sulfate and evaporated to dryness.
ESI(+)MS：m/z647(MH+)。
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-4- [ (1-methylpiperidin-4-yl) amino]-2- [ tetrahydro-2H-pyran-4-ylamino]Benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (1-methylpiperidin-4-yl) amino]-2- [ tetrahydro-2H-pyran-4-ylamino]-phenyl radical]cpd.108
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.28-1.50(m，4H)1.80-1.99(m，4H)2.06(t，J=12.54Hz，2H)2.19(s，3H)2.75(d，J=12.19Hz，2H)3.40(m，1H)3.45(ddd，J=11.83，10.12，2.32Hz，2H)3.83(dt，J=11.68，3.86Hz，2H)4.03(s，2H)5.87(d，J=1.71Hz，1H)5.90(dd，J=8.78，1.95Hz，1H)5.93(d，J=7.93 Hz，1H)5.95(s，1H)6.98(m,3H)7.24(dd，J=8.66，1.59Hz，1H)7.39(d，J=8.54Hz，1H)7.47(br.s.，1H)7.69(d，J=8.90Hz，1H)8.30(d，J=7.44Hz，1H)9.88(s，1H)12.57(s，1H)
4- [ (azetidin-3-ylmethyl) amino]-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-2- (tetrahydro-2H-pyran-4-ylamino) benzamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- [ (azetidin-3-ylmethyl) amino]-2- [ tetrahydro-2H-pyran-4-ylamino]Phenyl radical]Preparation of cpd.109
3- ({ [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) phenyl]Amino } methyl) azetidine-1-carboxylic acid tert-butyl ester (738mg, 1.1mmol) was dissolved in DCM (12mL) and TFA (3mL) was added. The resulting reaction solution was stirred at room temperature for 3 hours. The mixture was diluted with DCM and diluted with 10% NH3And (4) water extraction. The aqueous phase was extracted several times with DCM. The collected organic phases were washed with brine, dried over sodium sulfate and evaporated to dryness. Purification by column chromatography on silica gel using dichloromethane/methanol/NH in MeOH35N (70: 30: 1) as eluent, 150mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.42(m，2H)1.87-2.00(m，2H)2.77-2.88(m，1H)3.24-3.33(m，4H)3.42-3.53(m，2H)3.53-3.60(m，3H)3.78-3.88(m，2H)4.05(s，2H)5.86(s，1H)5.90(d，J=8.66Hz，1H)6.07-6.13(m，1H)6.95-7.04(m，3H)7.25(dd，J=8.60，1.52Hz，1H)7.40(d，J=8.66Hz，1H)7.48(s，1H)7.70(d，J=8.78Hz，1H)8.35(d，J=7.19Hz，1H)9.90(s，1H)12.59(br.s.，1H)。
Example 19
Step i'
Preparation of 2, 6-dichloro-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] pyridine-3-carboxamide
2, 6-dichloropyridine-3-carboxylic acid (480mg, 2.5mmol) and thionyl chloride (0.28mL, 3.75mmol) were heated in dry toluene (120mL) and several drops of dry DMF at 90 ℃ for 2 h. The volatiles were evaporated and the residue was dissolved in dry pyridine (15mL) at 0 ℃ under nitrogen. A solution of 5- (3, 5-difluoro-benzyl) -1H-indazol-3-ylamine (518mg, 2mmol) in dry pyridine (7mL) was added to the cooled reaction mixture. The resulting mixture was allowed to react at room temperature overnight, and then the solvent was removed under reduced pressure. The residue was dissolved in EtOAc and taken up with NaHCO3Saturated aqueous solution, water and brine. The organic phase was dried over sodium sulfate and evaporated to dryness. The crude product was purified by flash chromatography on silica gel with DCM/EtOH 100: 4 as eluent, 300mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.09(s，2H)6.93-7.01(m，2H)7.04(tt，J=9.39，2.32Hz，1H)7.29(dd，J=8.54，1.34Hz，1H)7.45(d，J=8.54Hz，1H)7.70(s，1H)7.75(d，J=8.05Hz，1H)8.24(d，J=7.93Hz，1H)11.04(s，1H)12.80(s，1H)
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -3, 5-difluoropyridine-2-carboxamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.07(s，2H)6.93-6.99(m，2H)6.99-7.06(m，1H)7.28(dd，J=8.66，1.46Hz，1H)7.45(d，J=8.41Hz，1H)7.68(s，1H)8.12-8.23(m，1H)8.68(s，1H)10.78(s，1H)12.81(s，1H)
Preparation of 6-chloro-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) pyridine-3-carboxamide
2, 6-dichloro-N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]Pyridine-3-carboxamide (80mg, 0.18mmol) in dioxaneA solution in an alkane (1mL) was heated at 100 deg.C for 24H in the presence of DIPEA (0.1mL, 0.55mmol) and tetrahydro-2H-pyran-4-amine (28mg, 0.28 mmol). The reaction mixture was diluted with EtOAc and washed with water. The organic layer was dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel with DCM/EtOH 95: 5 as eluent, 57mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.37-1.52(m，2H)1.94(dd，J=13.05，2.80Hz，2H)3.47(td，J=11.16，2.19Hz，2H)3.80-3.87(m，2H)4.06(s，2H)4.07-4.15(m，1H)6.73(d，J=8.05Hz，1H)6.93-7.07(m，3H)7.28(dd，J=8.66，1.59Hz，1H)7.44(dd，J=8.54，0.49Hz，1H)7.55(s，1H)8.29(d，J=8.17Hz，1H)8.60(d，J=7.32Hz，1H)10.74(s，1H)12.79(s，1H)
The following compounds were obtained by working in the same manner as described above:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] -5-fluoro-3- (tetrahydro-2H-pyran-4-ylamino) pyridine-2-carboxamide
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34-1.52(m，2H)1.95(d，J=10.36Hz，2H)3.45-3.54(m，2H)3.68-3.77(m，1H)3.82-3.89(m，2H)4.07(s，2H)6.97-7.05(m，3H)7.28(dd，J=8.66，1.59Hz，1H)7.37(dd，J=12.44，2.32Hz，1H)7.43(d，J=8.54Hz，1H)7.65(s，1H)7.88(d，J=2.32Hz，1H)8.55(d，J=6.95Hz，1H)10.46(s，1H)12.76(s，1H)
N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-5- (4-methylpiperazin-1-yl) -3- (tetrahydro-2H-pyran-4-ylamino) pyridine-2-carboxamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =5- (4-methylpiperazin-1-yl) -3- (tetrahydro-2H-pyran-4-ylamino) pyridine]Preparation of cpd.113
Reacting N- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]A solution of-5-fluoro-3- (tetrahydro-2H-pyran-4-ylamino) pyridine-2-carboxamide (925mg, 1.92mmol) and N-methylpiperazine (20mL) was stirred at 60 ℃ for 48 hours. The reaction mixture was then diluted with EtOAc and saturated NaHCO3And (4) washing the solution. The organic layer was dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel using DCM/EtOH/NH in MeOH35N (100: 5: 0.5) as eluent, 600mg of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34-1.47(m，2H)1.92-2.00(m，2H)2.25(s，3H)2.44-2.49(m，4H)3.34-3.40(m，4H)3.48-3.56(m，2H)3.72-3.81(m，1H)3.82-3.88(m，2H)4.07(s，2H)6.54(d，J=2.20Hz，1H)6.95-7.07(m，3H)7.26(dd，J=8.66，1.59Hz，1H)7.41(d，J=8.54Hz，1H)7.72(s，1H)7.73(d，J=2.32Hz，1H)8.32(d，J=8.05Hz，1H)10.19(s，1H)12.66(s，1H)
The following compounds were obtained by operating in the same manner:
n- [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl]-6- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) pyridine-3-carboxamide [ (I)A) R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =6- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) pyridine]cpd.114
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.35-1.47(m，2H)1.90-2.00(m，2H)2.22(s，3H)2.36-2.40(m，4H)3.41-3.51(m，2H)3.57-3.63(m，4H)3.78-3.88(m，2H)4.05(s，2H)4.06-4.11(m，1H)6.10(d，J=8.90Hz，1H)6.96-7.05(m，3H)7.25(dd，J=8.66，1.59Hz，1H)7.41(d，J=8.66Hz，1H)7.50(s，1H)8.10(d，J=9.02Hz，1H)8.73(d，J=6.95Hz，1H)10.06(s，1H)12.63(s，1H)。
Example 20
Step v
Preparation of tert-butyl 4- [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) phenyl ] piperazine-1-carboxylate
To a solution of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4-piperazin-1-yl-2- (tetrahydro-pyran-4-ylamino) -benzamide (71.7mg, 0.131mmol) in anhydrous dichloromethane (3.0mL) and triethylamine (0.052mL, 38.1mg, 0.377mmol) was added di-tert-butyl dicarbonate (34.5mg, 0.157mmol), and the solution was stirred at room temperature for 40 minutes. The mixture was evaporated to dryness and purified by flash silica gel column chromatography, eluting with dichloromethane/methanol 9: 1 to yield 60mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.28-1.41(m，2H)1.44(s，9H)1.90-1.99(m，2H)3.24-3.30(m，4H)3.46(d，J=4.88Hz，4H)3.48-3.54(m，2H)3.64-3.74(m，1H)3.79-3.86(m，2H)4.05(s，2H)6.16(d，J=2.19Hz，1H)6.25(dd，J=8.90，2.19Hz，1H)6.95-7.04(m，3H)7.26(dd，J=8.66，1.46Hz，1H)7.41(d，J=8.90Hz，1H)7.49(s，1H)7.82(d，J=9.15Hz，1H)8.29(d，J=7.44Hz，1H)10.10(s，1H)12.64(s，1H)
Preparation of ethyl 5- (3, 5-difluorobenzyl) -3- ({ [4- (piperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl ] carbonyl } amino) -1H-indazole-1-carboxylate
To a solution of tert-butyl 4- [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) phenyl ] piperazine-1-carboxylate (0.013mmol) in anhydrous tetrahydrofuran (1.0mL) maintained at-50 ℃ was added a 1M solution of LiHMSD in anhydrous tetrahydrofuran (0.015mL) under an argon atmosphere. After stirring at this temperature for 5 minutes, ethyl chlorocarbonate (0.002mL, 1.63mg, 0.015mmol) was added. After 30 minutes at-50 ℃, the reaction was complete. After dilution with dichloromethane, the solution was washed with brine, dried over sodium sulfate and evaporated to dryness. The crude product was dissolved in dichloromethane (1mL), trifluoroacetic acid (0.1mL) was added, and the mixture was stirred at room temperature overnight. After dilution with dichloromethane, the solution was washed with sodium bicarbonate, brine, dried over sodium sulfate and evaporated to dryness.
The crude product was purified by flash chromatography on silica gel with dichloromethane/methanol 9: 1 and 0.5% of 33% NH4Aqueous OH was eluted to afford the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.38(m，2H)1.40(t，J=7.13Hz，3H)1.90-1.99(m，2H)2.79-2.86(m，4H)3.18 -3.23(m，4H)3.47-3.54(m，2H)3.63-3.76(m，1H)3.79-3.86(m，2H)4.11(s，2H)4.48(q，J=7.15Hz，2H)6.11(d，J=2.07Hz，1H)6.24(dd，J=9.15，2.19Hz，1H)6.97-7.07(m，3H)7.55(dd，J=8.66，1.59Hz，1H)7.67(d，J=0.73Hz，1H)7.80(d，J=9.02Hz，1H)8.07(d，J=8.66Hz，1H)8.24(d，J=7.56Hz，1H)10.65(br.s.，1H)
Preparation of 1- (acetoxy) ethyl 4- [4- { [5- (3, 5-difluorobenzyl) -1H-indazol-3-yl ] carbamoyl } -3- (tetrahydro-2H-pyran-4-ylamino) phenyl ] piperazine-1-carboxylate
N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl cooled to 0 ℃ in a nitrogen atmosphere]To a solution of-4-piperazin-1-yl-2- (tetrahydro-pyran-4-ylamino) -benzamide in chloroform (5.0mL) were added 1, 8-bis (dimethylamino) naphthalene (21.4mg, 0.1mmol) and (1-chloroethyl) chloroformate (0.011mL, 14.3mg, 0.1 mmol). After stirring at room temperature for 2h, the mixture was diluted with dichloromethane (30mL), washed with saturated sodium bicarbonate solution (3mL), brine (3 × 5mL), dried over sodium sulfate and evaporated to dryness. The crude product was dissolved in glacial acetic acid (2.0mL), mercury (II) acetate (31.9mg, 0.1mmol) was added, and the mixture was stirred at room temperature for 1.5 h. After removal of the solvent, the crude product was dissolved in dichloromethane, washed with saturated sodium bicarbonate solution (3 × 3mL), brine (3 × 5mL), dried over sodium sulfate and evaporated to dryness to give 50mg of a light yellow foam which was purified by flash chromatography on silica gel with ethyl acetate and 0.5% of 33% NH4Aqueous OH was eluted to give 35mg of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.29-1.42(m，2H)1.46(d，J=5.49Hz，3H)1.90-1.98(m，2H)2.03-2.06(m，3H)3.30-3.50(m，8H)3.45-3.52(m，2H)3.64-3.74(m，1H)3.79-3.86(m，2H)4.05(s，2H)6.16(d，J=2.07Hz，1H)6.25(dd，J=9.02，2.07Hz，1H)6.67-6.73(m，1H)6.94-7.05(m，3H)7.26(dd，J=8.66，1.59Hz，1H)7.41(d，J=8.66Hz，1H)7.49(s，1H)7.82(d，J=9.02Hz，1H)8.30(d，J=7.68Hz，1H)10.11(s，1H)12.64(s，1H)
Preparation of ethyl 5- (3, 5-difluorobenzyl) -3- ({ [4- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl ] carbonyl } amino) -1H-indazole-1-carboxylate [ (XXVII), R1= R2= R3= H, R =3, 5-difluorophenyl, Ar =4- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl, PG = ethoxycarbonyl ] cpd.140
To a solution of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide (200mg, 0.356mmol) in anhydrous tetrahydrofuran (9mL), maintained at-50 ℃ and under a nitrogen atmosphere, was added a 1M solution of LiHMSD in anhydrous tetrahydrofuran (0.374 mL). After stirring at this temperature for 5 minutes, ethyl chloroformate (0.036mL, 0.374mmol) was added. After 1 hour at-50 ℃, the reaction was complete. The reaction mixture was diluted with water/EtOAc, washed with brine, dried over sodium sulfate and evaporated to dryness. By using DCM/ethanol 100: 5 flash chromatography on silica gel to give 140mg (62% yield) of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.39(t，J=7.07Hz，3H)2.25(br.s.，3H)2.46(br.s.，4H)3.50(ddd，J=11.83，10.06，2.26Hz，1H)3.66-3.75(m，1H)3.81(dt，J=11.61，3.76Hz，2H)4.10(s，2H)4.47(q，J=7.15Hz，2H)6.13(d，J=1.95Hz，1H)6.25(dd，J=9.08，2.13Hz，1H)7.54(dd，J=8.66，1.59Hz，1H)7.66(dd，J=1.46，0.73Hz，1H)7.80(d，J=9.15Hz，1H)8.07(d，J=8.66Hz，1H)8.24(d，J=7.68Hz，1H)10.65(s，1H)
The following compounds were obtained by working in the same manner as described above:
2-methoxyethyl 5- (3, 5-difluorobenzyl) -3- ({ [4- (4-methylpiperazin-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) phenyl ] carbonyl } amino) -1H-indazole-1-carboxylate
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.30-1.43(m，2H)1.90-2.00(m，2H)2.26(br.s.，3H)2.47(br.s.，4H)3.27-3.33(m，7H)3.46-3.55(m，2H)3.67-3.74(m，3H)3.79-3.85(m，2H)4.11(s，2H)4.54-4.59(m，2H)6.14(d，J=1.71Hz，1H)6.26(dd，J=9.02，2.19Hz，1H)6.97-7.09(m，3H)7.56(dd，J=8.72，1.52Hz，1H)7.67(d，J=0.85Hz，1H)7.81(d，J=9.15Hz，1H)8.07(d，J=8.54Hz，1H)8.25(d，J=7.56Hz，1H)10.68 (s，1H)
5- (3, 5-Difluorobenzyl) -3- [ ({4- [4- (ethoxycarbonyl) piperazin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) phenyl } carbonyl) amino ] -1H-indazole-1-carboxylic acid ethyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.22(t，J=7.07Hz，3H)1.30-1.38(m，2H)1.40(t，J=7.07Hz，3H)1.90-2.00(m，2H)3.48-3.54(m，2H)3.71(d，1H)3.78-3.86(m，2H)4.05-4.10(m，2H)4.11(s，2H)4.48(q，J=7.03Hz，2H)6.15(d，J=2.07Hz，1H)6.26(dd，J=9.15，2.19Hz，1H)6.95-7.07(m，2H)7.55(dd，J=8.66，1.59Hz，1H)7.67(d，J=0.85Hz，1H)7.83(d，J=9.15Hz，1H)8.08(d，J=8.78Hz，1H)8.25(d，J=7.80Hz，1H)10.68(s，1H)
Example 21
Preparation of 4-fluoro-2-nitro-benzoic acid tert-butyl ester
4-fluoro-2-nitrobenzoic acid (10g, 54mmol), (Boc)2A solution of O (2eq., 23.6g, 108mmol) and 4- (N, N-dimethylamino) pyridine (0.3eq., 1.98g, 16.2mmol) in tert-butanol (100mL) and dichloromethane (100mL) was stirred at room temperature for 20 hours. The reaction mixture was then diluted with ethyl acetate (500mL), washed with 1N HCl (500mL), water (500mL), brine (500mL), dried over sodium sulfate and evaporated to dryness. The title compound was obtained as a light yellow oil (quantitative) which was used in the next step without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：8.04(dd，J=8.47，2.50Hz，1H)7.95(dd，J=8.66，5.37Hz，1H)7.71(ddd，J=8.66，8.17，2.56Hz，1H)1.51(s，9H)。
Preparation of 4- (4-methyl-piperazin-1-yl) -2-nitro-benzoic acid tert-butyl ester
A solution of 4-fluoro-2-nitro-benzoic acid tert-butyl ester (13g, 54mmol) and N-methylpiperazine (17mL) was stirred at room temperature for 6 hours. The reaction mixture was then diluted with water (800mL) and maintained under magnetic stirring for 20 hours. The resulting solid was filtered, washed well with water and dried in vacuo at 40 ℃. The title compound was obtained as a yellow solid (16.4g, 94% yield) which was used in the next step without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.69(d，J=8.90Hz，1H)7.29(d，J=2.56Hz，1H)，7.15(dd，J1=8.90Hz，J2=2.56Hz，1H)，3.37(m，4H)，2.44(m，4H)，1.46(s，9H)。
The following compounds were obtained by operating in the same manner:
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2-nitro-benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.67(d，J=8.9Hz，1H)，6.98(d，J=2.6Hz，1H)，6.89(dd，J1=8.9Hz，J2=2.6Hz，1H)，3.54(m，2H)，3.02(s，3H)，2.40(m，2H)，2.19(s，6H)，1.46(s，9H)。
4- (4-dimethylamino-piperidin-1-yl) -2-nitro-benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.67(d，J=9.0Hz，1H)，7.26(d，J=2.6Hz，1H)，7.13(dd，J1=9.0Hz，J2=2.6Hz，1H)，3.96(m，2H)，2.93(m，2H)，2.36(m，1H)，2.20(s，6H)，1.82(m，2H)，1.46(s，9H)，1.40(m，2H)。
4- [ (3-dimethylamino-propyl) -methyl-amino ] -2-nitro-benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.67(d，J=9.0Hz，1H)，7.02(d，J=2.6Hz，1H)，6.90(dd，J1=9.0Hz，J2=2.6Hz，1H)，3.46(m，2H)，3.00(s，3H)，2.22(m，2H)，2.14(s，6H)，1.65(m，2H)，1.45(s，9H)。
4- (4-methyl-1, 4-diazepan-1-yl) -2-nitrobenzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.44(s，9H)1.85(m，2H)2.25(s，3H)2.43(m，2H)2.60(m，2H)3.51(t，2H)3.60(t，2H)6.91(dd，J1=9.02Hz，J2=2.66Hz，1H)7.02(d，J=2.56Hz，1H)7.64(d，J=8.90Hz，1H)
2-Nitro-4- (piperazin-1-yl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.46(m，9H)2.81(m，4H)3.33(m，4H)7.12(dd，J1=8.90Hz，J2=2.56Hz，1H)7.25(d，J=2.56Hz，1H)7.65(d，J=8.90Hz，1H)
2-Nitro-4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] benzoic acid tert-butyl ester
ESI(+)MS：m/z376(MH+)。
Preparation of 2-amino-4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester
A mixture of 4- (4-methyl-piperazin-1-yl) -2-nitro-benzoic acid tert-butyl ester (13.3g, 41.5mmol), cyclohexane (45mL), ethanol (300mL) and 10% Pd/C (0.4g) was stirred at 80 ℃ for 7 hours. An additional 10% Pd/C (0.9g) was added and the mixture was stirred at 80 ℃ for an additional 4 hours. The reaction mixture was filtered through a pad of celite, washed well with ethanol and the filtrate evaporated to dryness to give the title compound as a pale yellow solid (11.5g, 95% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.47(d，J=9.0Hz，1H)，6.40(bs，2H)，6.18(dd，J1=9.0Hz，J2=2.4Hz，1H)，6.11(d，J=2.4Hz，1H)，3.16(m，4H)，2.41(m，4H)，2.21(s，3H)，1.49(s，9H)。
The following compounds were obtained by operating in the same manner:
2-amino-4- [ (2-dimethylamino-ethyl) -methyl-amino ] -benzoic acid tert-butyl ester
ESI(+)MS：m/z294(MH+)。
2-amino-4- [ (3-dimethylamino-propyl) -methyl-amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.45(d，J=9.0Hz，1H)，6.36(bs，2H)，5.99(dd，J1=9.0Hz，J2=2.6Hz，1H)，5.86(d，J=2.6Hz，1H)，3.31(m，2H)，2.87(s，3H)，2.22(m，2H)，2.15(s，6H)，1.62(m，2H)，1.48(s，9H)。
2-amino-4- [4- (trifluoroacetyl) piperazin-1-yl ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.51(s，9H)3.28-3.35(m，4H)3.66-3.74(m，4H)6.15(d，J=2.44Hz，1H)6.21(dd，J=9.14，2.44Hz，1H)6.47(br.s.，2H)7.50-7.53(m，1H)
2-amino-4- [4- (dimethylamino) piperidin-1-yl ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.31-1.45(m，2H)1.49-1.52(m，9H)1.75-1.81(m，2H)2.17(s，6H)2.20-2.30(m，1H)2.69-2.79(m，2H)3.71-3.80(m，2H)6.12(d，J=2.44Hz，1H)6.18(dd，J=9.14，2.44Hz，1H)6.39(s，2H)7.46(d，J=9.02 Hz，1H)
2-amino-4- (4-methyl-1, 4-diazepan-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z306(MH+)。
2-amino-4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] benzoic acid tert-butyl ester
ESI(+)MS：m/z346(MH+)。
2-amino-4- (morpholin-4-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z279(MH+)。
Preparation of 4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzoic acid tert-butyl ester
To a solution of 2-amino-4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester (11.5g, 39.5mmol) in dichloromethane (340mL) was added tetrahydro-pyran-4-one (4.5mL, 49.3mmol), trifluoroacetic acid (8.2mL) and tetramethylammonium triacetoxyborohydride (15.57g, 59.2 mmol). The mixture was stirred at room temperature for 2 hoursThen with 0.5N hydrochloric acid, 0.5N NaOH and NaHCO3Washing with saturated solution. The organic layer was dried over sodium sulfate and evaporated to dryness to give the title compound as a pale yellow solid (13.3g, 90% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.72(d，J=7.7Hz，1H)，7.58(d，J=9.1Hz，1H)，6.20(dd，J1=9.1Hz，J2=2.2Hz，1H)，6.08(d，J=2.2Hz，1H)，3.85(m，2H)，3.70(m，1H)，3.50(m，2H)，3.27(m，4H)，2.47(m，4H)，2.26(bt，3H)，1.96(m，2H)，1.51(s，9H)，1.39(m，2H)。
The following compounds were obtained by operating in the same manner:
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- (tetrahydro-pyran-4-ylamino) -benzoic acid tert-butyl ester
ESI(+)MS：m/z378(MH+)。
4- [ (3-dimethylamino-propyl) -methyl-amino ] -2- (tetrahydro-pyran-4-ylamino) -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.70(bd，J=7.4Hz，1H)，7.54(d，J=9.0Hz，1H)，5.99(dd，J1=9.0Hz，J2=2.3Hz， 1H)，5.79(d，J=2.3Hz，1H)，3.86(m，2H)，3.62(m，1H)，3.47(m，2H)，3.36(m，2H)，2.93(s，3H)，2.28(m，2H)，2.18(bs，6H)，1.97(m，2H)，1.64(m，2H)，1.49(s，9H)，1.39(m，2H)。
2- (tetrahydro-2H-pyran-4-ylamino) -4- [4- (trifluoroacetyl) piperazin-1-yl ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.33-1.45(m，2H)1.51(s，9H)1.92-2.00(m，2H)3.36-3.42(m，4H)3.50(td，J=11.18，2.13Hz，2H)3.70(d，J=3.05Hz，5H)3.82-3.89(m，2H)6.10(d，J=2.32Hz，1H)6.21(dd，J=9.08，2.26Hz，1H)7.61(d，J=9.02Hz，1H)7.73(d，J=7.68Hz，1H)
4- [4- (dimethylamino) piperidin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z404(MH+)。
4- (4-methyl-1, 4-diazepan-1-yl) -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z390(MH+)。
4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] -2- (tetrahydro-2H-pyran-4-ylamino) benzoic acid tert-butyl ester
ESI(+)MS：m/z430(MH+)。
2- (cyclohexylamino) -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z374(MH+)。
2- [ (1, 3-Dimethoxypropan-2-yl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z394(MH+)。
2- (benzylamino) -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z382(MH+)。
4- (4-Methylpiperazin-1-yl) -2- { [ cis-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.40-1.50(m，2H)1.51 (s，9H)1.57-1.69(m，2H)1.70-1.78(m，2H)1.87(d，J=14.27Hz，2H)2.24(s，3H)2.32-2.39(m，1H)2.40-2.48(m，4H)3.27(br.s.，4H)3.83-3.94(m，1H)6.05(d，J=1.95Hz，1H)6.20(dd，J=9.21，2.26Hz，1H)7.57(d，J=9.02Hz，1H)8.04(d，J=8.05Hz，1H)
4- (4-Methylpiperazin-1-yl) -2- { [ trans-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.18-1.31(m，2H)1.44-1.57(m，2H)1.50(s，9H)1.87-1.94(m，2H)2.07-2.13(m，2H)2.25(s，3H)2.28-2.38(m，1H)2.44(br.s.，4H)3.26(br.s.，4H)3.40-3.53(m，1H)6.07(d，J=2.07Hz，1H)6.18(dd，J=9.08，2.26Hz，1H)7.54-7.58(m，1H)7.62(d，J=7.93Hz，1H)
4- (4-Methylpiperazin-1-yl) -2- ({ cis-4- [ (phenylcarbonyl) oxy ] cyclohexyl } amino) benzoic acid tert-butyl ester
ESI(+)MS：m/z494(MH+)。
4- (4-Methylpiperazin-1-yl) -2- ({ trans-4- [ (phenylcarbonyl) oxy ] cyclohexyl } amino) benzoic acid tert-butyl ester
ESI(+)MS：m/z494(MH+)。
2- [ (1-Methylpiperidin-4-yl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z291(MH+)。
2- [ (1-Methylpiperidin-4-yl) amino ] -4- (morpholin-4-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z376(MH+)。
Preparation of 4- (4-methyl-piperazin-1-yl) -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
To a solution of 4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzoic acid tert-butyl ester (13.3g, 35.4mmol) in dry dichloromethane (350mL) under an argon atmosphere and at 0 ℃ was added triethylamine (7.5mL, 53.1mmol) and trifluoroacetic anhydride (6.5mL, 46.1 mmol). The mixture was stirred at 0 ℃ for 20 minutes, then water (350mL) was added dropwise. The phases were separated and the organic phase was washed with brine, dried over sodium sulfate and evaporated to dryness. The crude residue was purified by silica gel chromatography with dichloromethane/ethanol 95: 5 as eluent, 12.1g of the title compound was obtained as a pale yellow solid (73% yield).
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.83(d，J=9.0Hz，1H)，7.06(dd，J1=9.0Hz，J2=2.5Hz，1H)，6.82(J=2.5Hz，1H)，4.48(m，1H)，3.85(m，2H)，3.5-3.3(m，6H)，2.49(m，4H)，2.26(bs，3H)，2.0(m，1H)，1.59(m，1H)，1.51(m，1H)，1.46(s，9H)，1.03(m，1H)。
The following compounds were obtained by operating in the same manner:
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.80(d，J=9.1Hz，1H)，6.79(dd，J1=9.1Hz，J2=2.6Hz，1H)，6.51(d，J=2.6Hz，1H)，4.48(m，1H)，3.86(m，1H)，3.79(m，1H)，3.52(m，2H)，3.41-3.25(m，2H)，3.00(s，3H)，2.5-2.35(m，2H)，2.21(s，6H)，1.98(m，1H)，1.64-1.45(m，3H)，1.44(s，9H)。
4- [ (3-dimethylamino-propyl) -methyl-amino ] -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.79(d，J=9.1Hz，1H)，6.79(dd，J1=9.1Hz，J2=2.6Hz，1H)，6.52(d，J=2.6Hz，1H)，4.48(m，1H)，3.87(m，1H)，3.79(m，1H)，3.51-3.32(m，4H)，2.98(s，3H)，2.22(m，2H)，2.12(s，6H)，1.99(m，1H)，1.70-1.46(m，4H)，1.44(s，9H)，1.03(m，1H)。
2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] -4- [4- (trifluoroacetyl) piperazin-1-yl ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.45(s，9H)1.60(qd，J=12.21，4.94Hz，2H)3.73(t，J=5.12Hz，4H)4.48(tt，J=11.96，3.89Hz，1H)6.84(d，J=2.56Hz，1H)7.07(dd，J=8.96，2.62Hz，1H)7.85(d，J=9.02Hz，1H)
4- [4- (dimethylamino) piperidin-1-yl ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z500(MH+)。
4- (4-methyl-1, 4-diazepan-1-yl) -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z486(MH+)。
4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z526(MH+)。
2- [ cyclohexyl (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z470(MH+)。
2- [ (1, 3-Dimethoxypropan-2-yl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z490(MH+)。
2- [ benzyl (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z478(MH+)。
4- (4-Methylpiperazin-1-yl) -2- { (trifluoroacetyl) [ cis-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid tert-butyl ester
ESI(+)MS：m/z538(MH+)。
4- (4-Methylpiperazin-1-yl) -2- { (trifluoroacetyl) [ trans-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid tert-butyl ester
ESI(+)MS：m/z538(MH+)。
4- (4-Methylpiperazin-1-yl) -2- [ { cis-4- [ (phenylcarbonyl) oxy ] cyclohexyl } (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z590(MH+)。
4- (4-Methylpiperazin-1-yl) -2- [ { trans-4- [ (phenylcarbonyl) oxy ] cyclohexyl } (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z590(MH+)。
2- [ (1-Methylpiperidin-4-yl) (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z387(MH+)。
2- [ (1-Methylpiperidin-4-yl) (trifluoroacetyl) amino ] -4- (morpholin-4-yl) benzoic acid tert-butyl ester
ESI(+)MS：m/z472(MH+)。
4- (4-Methylpiperazin-1-yl) -2- [ phenyl (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z464(MH+)。
Preparation of 4- (4-methyl-piperazin-1-yl) -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid trifluoroacetate salt
A mixture of 4- (4-methyl-piperazin-1-yl) -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester (12.1g, 25.7mmol), trifluoroacetic acid (48.5mL) and dichloromethane (195mL) was stirred at room temperature for 2 hours. The volatiles were then evaporated, the residue was dissolved in ether and evaporated again. This procedure was repeated 5 times, then the solid was triturated with ether, filtered and dried in an oven at 40 ℃ to give the title compound as a beige solid (13.4 g).
1H-NMR(400MHz)，(ppm，DMSO-d6)：12.78(bs，1H)，9.74(bs，1H)，7.93(d，J=8.8Hz，1H)，7.13(dd，J1=8.8Hz，J2=2.5Hz，1H)，6.98(d，J=2.5Hz，1H)，4.49(m，1H)，4.11(m，2H)，3.84(m，2H)，3.6-3.0(m，8H)，2.89(s，3H)，1.98(m，1H)，1.59(m，1H)，1.53(m，1H)，1.08(m，1H)。
The following compounds were obtained by operating in the same manner:
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid trifluoroacetate salt
1H-NMR(400MHz)，(ppm，DMSO-d6)：12.56(bs，1H)，9.49(bs，1H)，7.88(d，J=8.9Hz，1H)，8.92(dd，J1=8.9Hz，J2=2.6Hz，1H)，6.63(d，J=2.6Hz，1H)，4.49(m，1H)，3.9-3.2(m，8H)，3.02(s，3H)，2.85(s，6H)，1.98(m，1H)，1.62-1.49(m，2H)，1.08(m，1H)。
4- [ (3-dimethylamino-propyl) -methyl-amino ] -2- [ (tetrahydro-pyran-4-yl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid trifluoroacetate salt
ESI(+)MS：m/z432(MH+)。
2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] -4- [4- (trifluoroacetyl) piperazin-1-yl ] benzoic acid
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.08(m，J=12.35，12.24，12.24，4.76Hz，1H)1.47-1.55(m，1H)1.56-1.67(m，1H)1.91-2.01(m，1H)3.38-3.53(m)3.73(t，J=5.12Hz，4H)3.78(dd，J=11.52，4.45Hz，1H)3.86(dd，J=11.40，4.57Hz，1H)4.46(tt，J=11.87，3.98Hz，1H)6.85(d，1H)7.06(dd，J=8.90，2.68Hz，1H)7.89(d，J=8.90Hz，1H)12.67(br.s.，1H)
4- (4-methyl-1, 4-diazepan-1-yl) -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：4.42-4.55(m，1H)6.91-6.96(m，1H)7.89(d，J=9.02Hz，1H)10.14(br.s.，1H)12.56(br.s.，1H)
4- [4- (dimethylamino) piperidin-1-yl ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z444(MH+)。
4- [ (2S) -2- (pyrrolidin-1-ylmethyl) pyrrolidin-1-yl ] -2- [ tetrahydro-2H-pyran-4-yl (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z470(MH+)。
2- [ cyclohexyl (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
ESI(+)MS：m/z414(MH+)。
2- [ (1, 3-Dimethoxyprop-2-yl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
ESI(+)MS：m/z434(MH+)。
2- [ benzyl (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
ESI(+)MS：m/z422(MH+)。
Trifluoroacetic acid salt of 4- (4-methylpiperazin-1-yl) -2- { (trifluoroacetyl) [ cis-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.09-1.90(4m，8H)2.36-2.46(m，1H)2.88(br.s.，3H)2.99-3.25(m，4H)3.49(br.s.，2H)3.96-4.16(m，2H)4.27-4.37(m，1H)7.00(d，J=2.32Hz，1H)7.12(dd，J=8.90，2.44Hz，1H)7.92(d，J=8.90Hz，1H)9.67(br.s.，1H)12.80(s，1H)
Trifluoroacetic acid salt of 4- (4-methylpiperazin-1-yl) -2- { (trifluoroacetyl) [ trans-4- (trifluoromethyl) cyclohexyl ] amino } benzoic acid
ESI(+)MS：m/z482(MH+)。
4- (4-methylpiperazin-1-yl) -2- [ { cis-4- [ (phenylcarbonyl) oxy ] cyclohexyl } (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z534(MH+)。
4- (4-methylpiperazin-1-yl) -2- [ { trans-4- [ (phenylcarbonyl) oxy ] cyclohexyl } (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z534(MH+)。
2- [ (1-Methylpiperidin-4-yl) (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z331(MH+)。
2- [ (1-Methylpiperidin-4-yl) (trifluoroacetyl) amino ] -4- (morpholin-4-yl) benzoic acid hydrochloride
ESI(+)MS：m/z416(MH+)。
4- (4-Methylpiperazin-1-yl) -2- [ phenyl (trifluoroacetyl) amino ] benzoic acid hydrochloride
ESI(+)MS：m/z408(MH+)。
Example 22
Preparation of 2, 4-difluoro-benzoic acid tert-butyl ester
To a solution of 2, 4-difluorobenzoic acid (5g, 31.62mmol) in a mixture of dichloromethane (100mL) and t-BuOH (50mL) was added (Boc)2O (13.8g, 63.24mmol) and N, N-dimethylaminopyridine (1.16g, 9.49 mmol). The solution was stirred at room temperature for 24 hours, then diluted with dichloromethane, 1N HCl, NaHCO3The saturated solution, water (3 times) and brine were washed twice. The organic phase was dried over sodium sulfate, filtered and evaporated to give the title compound (5.70g, 84%) as a light yellow oil.
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.91(m，1H)，7.36(m，1H)，7.20(m，1H)，1.53(s，9H)。
Preparation of 4-fluoro-2- ((S) -2-methoxy-1-methyl-ethylamino) -benzoic acid tert-butyl ester
A mixture of tert-butyl 2, 4-difluoro-benzoate (30g, 140.05mmol) and (S) -2-methoxy-1-methyl-ethylamine (100mL) was stirred at 65 ℃ for 2 days. Adding NaHCO3Saturated solution ofAnd the mixture was extracted with dichloromethane (3 times). The organic phase was washed twice with water and then brine, dried over sodium sulfate, filtered and evaporated to dryness to give the crude product, which was purified by silica gel column chromatography (hexane/ethyl acetate 9: 1). The title compound (33.38g, 84%) was obtained as an oil.
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.87(d，J=7.80Hz，1H)，7.80(t，J=7.19Hz，1H)，6.60(dd，J1=13.05Hz，J2=2.44Hz，1H)，6.36(m，1H)，3.80(m，1H)，3.40(d，J=4.76Hz，2H)，3.30(s，3H)，1.53(s，9H)，1.17(d，J=6.58Hz，3H)。
The following compounds were obtained by working in the same manner as described above:
4-fluoro-2- ((R) -2-methoxy-1-methyl-ethylamino) -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.87(d，J=7.80Hz，1H)，7.80(t，J=7.19Hz，1H)，6.60(dd，J1=13.05Hz，J2=2.44Hz，1H)，6.36(m，1H)，3.80(m，1H)，3.40(d，J=4.76Hz，2H)，3.30(s，3H)，1.53(s，9H)，1.17(d，J=6.58Hz，3H)。
4-fluoro-2- (2-methoxy-ethylamino) -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.89(t，J=5.00Hz，1H)，7.80(t，J=7.07Hz，1H)，6.56(dd，J1=12.80Hz，J2=2.56Hz，1H)，6.37(m，1H)，3.55(t，J=5.37Hz，2H)，3.33(m，2H)，3.29(s，3H)，1.53(s，9H)。
4-fluoro-2- [ (3-methoxypropyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.51-1.53(m，9H)1.76 -1.85(m，2H)3.18-3.23(m，2H)3.25(s，3H)3.38-3.44(m，2H)6.32-6.39(m，1H)6.49(dd，J=12.80，2.44Hz，1H)7.79(dd，J=8.90，7.07Hz，1H)7.88(br.s.，1H)
4-fluoro-2- [ (2-fluoroethyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.54(s，9H)3.50(dd，J=27.00，5.00Hz，2H)4.63(dt，J=47.56，4.88Hz，2H)6.41(td，J=8.57，2.50Hz，1H)6.62(dd，J=12.62，2.38Hz，1H)7.82(dd，J=8.90，7.07Hz，1H)8.05(t，J=4.82Hz，1H)
4-fluoro-2- [ (3-fluoropropyl) amino ] benzoic acid tert-butyl ester
ESI(+)MS：m/z272(MH+)。
4-fluoro-2- [ (1-methoxy-2-methylprop-2-yl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.34(s，6H)1.53(s，9H)3.33(br.s.，3H)3.40(s，2H)6.31-6.39(m，1H)6.67(dd，J=13.29，2.44Hz，1H)7.82(dd，J=8.84，7.38Hz，1H)8.22(s，1H)
Preparation of 4-fluoro-2- [ ((S) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
A solution of 4-fluoro-2- ((S) -2-methoxy-1-methyl-ethylamino) -benzoic acid tert-butyl ester (1.54g, 5.44mmol) in dichloromethane (30mL) was cooled to 0 ℃ -5 ℃. Triethylamine (1.11mL, 8.16mmol) and trifluoroacetic anhydride (1.15mL, 8.16mmol) were added. After 3 hours at 0 ℃ to 5 ℃ with NaHCO3The mixture was washed with saturated solution, water and brine. The organic layer was dried over sodium sulfate, filtered and evaporated to give the title compound as a light yellow oil (2g, 99%).
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 8.07(m, 1H), 7.53(m, 1H), 7.29(dd, J1=9.39Hz, J2=2.68Hz, 1H), 4.83(m, 1H), 3.44(m, 1H), 3.30(s, 3H), 1.49(s, 9H), 0.86(d, 3H).
The following compounds were obtained by working in the same manner as described above:
4-fluoro-2- [ ((R) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 8.07(m, 1H), 7.53(m, 1H), 7.29(dd, J1=9.39Hz, J2=2.68Hz, 1H), 4.83(m, 1H), 3.44(m, 1H), 3.30(s, 3H), 1.49(s, 9H), 0.86(d, 3H).
4-fluoro-2- [ (2-methoxy-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：8.07(m，1H)，7.50(m，1H)，7.41(dd，J1=9.39Hz，J2=2.56Hz，1H)，4.28(m，1H)，3.55(m，1H)，3.46(m，1H)，3.38(m，1H)，3.18(s，3H)，1.49(s，9H)。
4-fluoro-2- [ (3-methoxypropyl) (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.48(s，9H)1.68-1.83(m，2H)3.18(s，3H)3.21-3.29(m，1H)3.33-3.38(m，2H)4.06-4.18(m，1H)7.46-7.52(m，1H)7.56(dd，J=9.27，2.68Hz，1H)8.06(dd，J=8.84，6.40Hz，1H)
4-fluoro-2- [ (2-fluoroethyl) (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.50(s，9H)3.54-3.74(m，1H)4.26-4.45(m，1H)4.50-4.80(m，2H)7.47-7.55(m，2H)8.08(dd，J=9.27，6.46Hz，1H)
4-fluoro-2- [ (3-fluoropropyl) (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.50(s，9H)1.80-2.07(m，2H)3.26-3.42(m，1H)4.21(ddd，J=13.78，8.90，6.71Hz，1H)4.42-4.60(m，2H)7.48-7.55(m，1H)7.60(dd，J=9.27，2.44Hz，1H)8.09(dd，J=8.84，6.40Hz，1H)
4-fluoro-2- [ (1-methoxy-2-methylpropan-2-yl) (trifluoroacetyl) amino ] benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.09(s，3H)1.47(s，3H)1.52(s，9H)3.17(s，3H)3.19(d，J=9.75Hz，1H)3.80(d， J=9.63Hz，1H)7.36(dd，J=9.45，2.62Hz，1H)7.47(td，J=8.41，2.68Hz，1H)7.93(dd，J=8.78，6.46Hz，1H)
Preparation of 2- [ ((S) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester
Reacting 4-fluoro-2- [ ((S) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino]A solution of tert-butyl benzoate (2g, 5.28mmol) and N-methylpiperazine (5.86mL, 52.8mmol) in tetrahydrofuran (20mL) was stirred at 60 ℃ for 7 days. The solution is then evaporated and NaHCO is added3The solution was saturated and the mixture was extracted with dichloromethane (3 times). The organic layer was washed with water, brine, dried over sodium sulfate, filtered and evaporated to give the crude product, which was purified by silica gel column chromatography (dichloromethane-methanol 93: 7). The title compound (2.04g, 84%) was obtained as a pale yellow solid.
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 7.81(d, J =9.15Hz, 1H), 7.06(dd, J1=9.15Hz, J2=2.56Hz, 1H), 6.79(d, J =2.56Hz, 1H), 4.80(m, 1H), 3.39(m, 2H), 3.34-3.28(m, 7H), 2.55(m, 4H), 2.29(bs, 3H), 1.46(s, 9H), 0.83(d, 3H).
The following compounds were obtained by working in the same manner as described above:
2- [ ((R) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 7.81(d, J =9.15Hz, 1H), 7.06(dd, J1=9.15Hz, J2=2.56Hz, 1H), 6.79(d, J =2.56Hz, 1H), 4.80(m, 1H), 3.39(m, 2H), 3.34-3.28(m, 7H), 2.55(m, 4H), 2.29(bs, 3H), 1.46(s, 9H), 0.83(d, 3H).
2- [ (2-methoxy-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 7.83(d, J =9.02Hz, 1H), 7.05(dd, J1=9.02Hz, J2=2.68Hz, 1H), 6.86(d, J =2.68Hz, 1H), 4.31(m, 1H), 3.55(m, 1H), 3.40(m, 1H), 3.32(m, 4H), 3.25(m, 1H), 3.21(s, 1H), 2.44(t, J =5.12Hz, 4H), 2.22(bs, 3H), 1.46(s, 9H).
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- [ (2-methoxy-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：7.81(d，J=8.9Hz，1H)，6.78(dd，J1=8.9Hz，J2=2.8Hz，1H)，6.60(d，J=2.8Hz，1H)，4.40-4.31(m，1H)，3.59-3.39(m，4H)，3.23(s，3H)，3.22-3.15(m，1H)，3.00(s，3H)，2.40(m，2H)，2.19(bs，6H)，1.46(s，9H)。
2- [ (3-methoxypropyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.45(s，9H)1.68-1.84(m，2H)2.26(br.s.，3H)2.44-2.60(m，4H)3.12-3.23(m，1H)3.18(s，3H)3.25-3.48(m，6H)4.08(d，J=22.92Hz，1H)6.92(d，J=2.19Hz，1H)7.02(dd，J=9.02，2.44Hz，1H)7.81(d，J=9.02Hz，1H)
2- [ (2-fluoroethyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.46(s，9H)2.22(s，3H)2.43(t，J=4.76Hz，4H)3.25-3.31(m，4H)3.41-3.59(m，1H)4.27-4.46(m，1H)4.46-4.78(m，2H)6.90(d，J=2.07Hz，1H)7.05(dd，J=9.02，2.68Hz，1H)7.83(d，J=9.02Hz，1H)
2- [ (3-Fluoropropyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.46(s，9H)1.80-2.05(m，2H)2.25(br.s.，3H)2.46(br.s.，4H)3.18-3.37(m，5H)4.10-4.24(m，1H)4.38-4.60(m，2H)6.95(d，J=2.44Hz，1H)7.04(dd，J=8.96，2.62Hz，1H)7.84(d，J=9.02Hz，1H)
2- [ (1-methoxy-2-methylpropan-2-yl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid tert-butyl ester
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.04(s，3H)1.45(s， 3H)1.49(s，9H)2.22(s，3H)2.44(t，J=4.94Hz，4H)3.20(d，J=9.51Hz，1H)3.23(s，3H)3.25-3.30(m，4H)3.93(d，J=9.51Hz，1H)6.89(d，J=2.32Hz，1H)7.00(dd，J=8.96，2.62Hz，1H)7.70(d，J=8.90Hz，1H)
Preparation of 2- [ ((S) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid trifluoroacetate
To a solution of 2- [ ((S) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid tert-butyl ester (2.03g, 4.42mmol) in dichloromethane (15mL) was added trifluoroacetic acid (3.4mL, 44.2 mmol). The mixture was stirred at room temperature for 15 hours, then the solution was evaporated to dryness to give the title compound as an oil, which was used in the next step without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 12.10(bs, 1H), 9.74(bs, 1H), 7.90(d, J =8.90Hz, 1H), 7.15(dd, J1=8.90Hz, J2=2.56Hz, 1H), 6.89(d, J =2.56Hz, 1H), 4.76(m, 1H), 4.03(t, 2H), 3.55(m, 2H), 3.37(m, 2H), 3.30(s, 3H), 3.18(m, 2H), 2.88(bs, 3H), 0.85(d, 3H).
The following compounds were obtained by working in the same manner as described above:
2- [ ((R) -2-methoxy-1-methyl-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid trifluoroacetate salt
1H-NMR(400MHz)，(ppm，DMSO-d6): (mixture of tautomers) 12.10(bs, 1H), 9.74(bs, 1H), 7.90(d, J =8.90Hz, 1H),7.15(dd，J1=8.90Hz，J2=2.56Hz，1H)，6.89(d，J=2.56Hz，1H)，4.76(m，1H)，4.03(t，2H)，3.55(m，2H)，3.37(m，2H)，3.30(s，3H)，3.18(m，2H)，2.88(bs，3H)，0.85(d，3H)。
2- [ (2-methoxy-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -4- (4-methyl-piperazin-1-yl) -benzoic acid trifluoroacetate salt
1H-NMR(400MHz)，(ppm，DMSO-d6): (tautomer mixture) 12.76(bs, 1H), 9.73(bs, 1H), 7.91(d, J =8.78Hz, 1H), 7.10(dd, J1=8.78Hz, J2=2.68Hz, 1H), 7.01(d, J =2.68Hz, 1H), 4.15(m, 1H), 4.04(m, 2H), 3.54(m, 2H), 3.42(m, 2H), 3.38(m, 2H), 3.33(m, 2H), 3.19(s, 3H), 3.14(m, 2H), 2.86(bs, 3H).
4- [ (2-dimethylamino-ethyl) -methyl-amino ] -2- [ (2-methoxy-ethyl) - (2,2, 2-trifluoro-acetyl) -amino ] -benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：12.59(bs，1H)，10.00(bs，1H)，7.88(d，J=8.9Hz，1H)，6.92(dd，J1=8.9Hz，J2=2.8Hz，1H)，6.74(8d，J=2.8Hz，1H)，4.18(m，1H)，3.79(m，2H)，3.56(m，1H)，3.47-3.36(m，2H)，3.24(m，2H)，3.21(s，3H)，3.01(s，3H)，2.84(bd，6H)。
2- [ (3-methoxypropyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.70-1.81(m，2H)2.84(d，J=2.93Hz，3H)3.06-3.40(m，7H)3.19(s，3H)3.52(d，J=10.36Hz，2H)3.96-4.06(m，1H)4.09(br.s.，2H)7.07(d，J=2.56Hz，1H)7.10(dd，J=8.90，2.68Hz，1H)7.93(d，J=8.78Hz，1H)10.27(br.s.，1H)12.76(br.s.，1H)
2- [ (2-fluoroethyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.84(br.s.，3H)3.04-3.30(m，4H)3.47-3.56(m，2H)3.54-3.67(m，1H)4.06(d，2H)4.18-4.40(m，1H)4.46-4.79(m，2H)7.07(d，J=2.19Hz，1H)7.12(dd，J=8.96，2.62Hz，1H)7.91-7.97(m，1H)10.33(br.s.，1H)12.83(br.s.，1H)
2- [ (3-Fluoropropyl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid trifluoroacetate salt
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.82-2.02(m，2H)2.87(s，3H)3.14(m，5H)3.44(m)4.09(m，3H)4.40-4.59(m，2H)7.08-7.15(m，2H)7.95(d，J=9.15Hz，1H)9.72(br.s.，1H)12.81(br.s.，1H)
2- [ (1-methoxy-2-methylpropan-2-yl) (trifluoroacetyl) amino ] -4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.07(s，3H)1.43(s，3H)2.84(s，3H)3.10-3.38(m，5H)3.25(s，3H)3.47-3.57(m，2H)3.92(d，J=9.51Hz，1H)3.95-4.02(m，2H)7.00(d，J=2.44Hz，1H)7.10(dd，J=8.84，2.50Hz，1H)7.84(d，J=8.78Hz，1H)10.25(br.s.，1H)12.77(br.s.，1H)
Example 23
Preparation of tert-butyl 4- (4-acetylpiperazin-1-yl) -2-nitrobenzoate
To a solution of tert-butyl 2-amino-4- (piperazin-1-yl) benzoate (7.6g, 24.7mmol) in dichloromethane (120mL) was added triethylamine (13.46mL, 98.7mmol) and trifluoroacetic anhydride (6.87mL, 49.35 mmol). After 1 hour the volatile material was evaporated and the crude product was purified by column chromatography (EtOAc/hexane 3: 7) to give 9.46g (yield 95%) of the title compound.
ESI(+)MS：m/z404(MH+)。
Example 24
Preparation of tert-butyl 4- (4-methylpiperazin-1-yl) -2- (phenylamino) benzoate
In a dry Schlenk tube under argon atmosphere, tert-butyl 2-amino-4- (4-methylpiperazin-1-yl) benzoate (800mg, 2.745mmol) was dissolved in dry toluene (14 mL). Argon was bubbled through the mixture for a few minutes, then bromobenzene (0.32mL, 3.02mmol, 1.1eq), Cs were added2CO2(1.34g，4.118mmol，1.5eq)，Pd(OAc)2(16mg, 0.069mmol, 2.5mol%) and Rac-BINAP (88mg, 0.137mmol, 5 mol%). The mixture was then stirred at 100 ℃ for 21 hours. The mixture was cooled to room temperature and diluted with dichloromethane. The salt was filtered through a pad of celite and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (DCM/EtOH/NH in methanol)37% = 95: 5: 0.5) to yield 1.13g of the title compound (quantitative yield) as an off-white solid.
1H-NMR(400MHz)，(ppm，DMSO-d6)：1.54(s，9H)2.21(s，3H)2.37-2.43(m，4H)3.15-3.20(m，4H)6.43(dd，J=9.15，2.44Hz，1H)6.60(d，J=2.44Hz，1H)7.02-7.07(m，1H)7.23 -7.27(m，2H)7.33-7.38(m，2H)7.69(d，J=9.02Hz，1H)9.50(s，1H)
Example 25
Preparation of methyl 2-methoxy-4- (4-methylpiperazin-1-yl) benzoate
2-methoxy-4-fluoro-benzoic acid methyl ester (1.6g, 9.7mmol), K2CO3(1.3g, 9.7mmol) and N-methylpiperazine (1.3mL, 11.7mmol) were heated in DMSO (5mL) at 100 ℃ for 20 h. The reaction mixture was diluted with DCM and washed with water. The organic phase was dried over sodium sulfate and evaporated to dryness. Purification by column chromatography on silica gel using dichloromethane/methanol 95: 5 as eluent, 1.7g (yield 66%) of the title compound were obtained.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.25(s，3H)2.45(br.s.，4H)3.26-3.34(m，4H)3.70(s，3H)3.80(s，3H)6.49(d，J=2.32Hz，1H)6.53(dd，J=8.84，2.38Hz，1H)7.61(d，J=8.78Hz，1H)
Preparation of 2-methoxy-4- (4-methylpiperazin-1-yl) benzoic acid hydrochloride
Methyl 2-methoxy-4- (4-methylpiperazin-1-yl) benzoate (1.9g, 7.2mmol) was heated at 40 ℃ in a mixture of 2N NaOH (10mL) and MeOH (10mL) for 2 hours. MeOH was evaporated and the aqueous layer was acidified with 25% HCl to pH =6, extracted with n-BuOH. The organic phase was dried over sodium sulphate and evaporated to dryness to yield 1.0g (61% yield) of the title compound.
1H-NMR(400MHz)，(ppm，DMSO-d6)：2.82(br.s.，3H)2.99-3.31(m，4H)3.47(br.s.，2H)3.83(s，3H)4.04(br.s.，2H)6.61(d，1H)6.59(s，1H)7.66(d，J=8.78Hz，1H)10.49(br.s.，1H)11.91(br.s.，1H)
Example 26
Preparation of 4- (4-methyl-piperazin-1-yl) -2-nitrobenzoate
4- (4-methyl-piperazin-1-yl) -2-nitro-benzoic acid tert-butyl ester (16.4g, 51mmol) and 37% HCl (100mL) in 1, 4-bisThe mixture in alkane (200mL) was stirred at room temperature for 4 hours. Filtering the obtained solid with 1, 4-bisThe alkane was washed well and dried under vacuum at 45 ℃. The title compound was obtained as a light yellow solid (13.45g, 87.5% yield), which was used in the next step without any further purification.
1H-NMR(400MHz)，(ppm，DMSO-d6)：10.27(bs，1H)，7.81(d，J=8.90Hz，1H)，7.40(d，J=2.69Hz，1H)，7.24(dd，J1=8.90Hz，J2=2.69Hz，1H)，4.13(bs，2H)，3.55-3.06(bs，6H)，2.83(s，3H)。
The following compounds were obtained by operating in the same manner:
4- [ (3-dimethylamino-propyl) -methyl-amino ] -2-nitro-benzoic acid hydrochloride
1H-NMR(400MHz)，(ppm，DMSO-d6)：13.07(bs，1H)，9.72(bs，1H)，7.76(d，J=9.0Hz，1H)，7.03(d，J=2.6Hz，1H)，6.93(dd，J1=9.0Hz，J2=2.6Hz，1H)，3.51(m，2H)，3.08(m，2H)，3.03(s，3H)，2.77(s，6H)，1.90(m，2H)。

Claims (11)

  1. Use of N- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (4-methyl-piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease caused by a dysregulated protein kinase activity.
  2. 2. The use of claim 1, wherein the disease is cancer.
  3. 3. The use of claim 2, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, prostate cancer, breast cancer, ovarian cancer, ewing's sarcoma, inflammatory myofibroblast tumor, anaplastic large cell lymphoma, rhabdomyosarcoma, renal cell carcinoma, thyroid follicular cancer, melanoma, myeloid hematopoietic tumors, lymphoid hematopoietic tumors, squamous cell carcinoma, gastric cancer, endometrial cancer, multiple myeloma, tumors of the central and peripheral nervous system, and retinoblastoma.
  4. 4. The use of claim 3, wherein the tumors of the central and peripheral nervous system are selected from the group consisting of neuroblastoma, glioblastoma, and medulloblastoma.
  5. 5. The following compounds or pharmaceutically acceptable salts thereof: n- [5- (3, 5-difluoro-benzyl) -1H-indazol-3-yl ] -4- (piperazin-1-yl) -2- (tetrahydro-pyran-4-ylamino) -benzamide.
  6. 6. Use of a compound of claim 5, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease caused by a dysregulated protein kinase activity.
  7. 7. The use of claim 6, wherein the disease is cancer.
  8. 8. The use of claim 7, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, prostate cancer, breast cancer, ovarian cancer, ewing's sarcoma, inflammatory myofibroblast tumors, anaplastic large cell lymphoma, rhabdomyosarcoma, renal cell carcinoma, thyroid follicular cancer, melanoma, myeloid hematopoietic tumors, lymphoid hematopoietic tumors, squamous cell carcinoma, gastric cancer, endometrial cancer, multiple myeloma, tumors of the central and peripheral nervous system, and retinoblastoma.
  9. 9. The use of claim 8, wherein the tumors of the central and peripheral nervous system are selected from the group consisting of neuroblastoma, glioblastoma, and medulloblastoma.
  10. 10. Use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from non-small cell lung cancer, colorectal cancer, prostate cancer, breast cancer, ovarian cancer, Ewing's sarcoma, inflammatory myofibroblast tumor, anaplastic large cell lymphoma, rhabdomyosarcoma, renal cell carcinoma, thyroid follicular cancer, melanoma, myeloid hematopoietic tumors, lymphoid hematopoietic tumors, squamous cell carcinoma, gastric cancer, endometrial cancer, multiple myeloma, central and peripheral nervous system tumors, and retinoblastoma,
    wherein:
    x is-CH2-；
    Ar is phenyl substituted with one or more substituents independently selected from NR5R6 and heterocyclyl, wherein:
    r5 and R6 are independently hydrogen, R8-O-C2-C6Alkyl radical, C3-C6Cycloalkyl, or heterocyclyl, or R5 and R6 together with the nitrogen atom to which they are bound may form an optionally straight or branched chain C1-C6An alkyl-substituted heterocyclic group;
    r8 is straight or branched chain C1-C6An alkyl group, a carboxyl group,
    r is phenyl optionally substituted with one or more halogens,
    r1, R2 and R3 are hydrogen,
    wherein said heterocyclyl is a 5-7 membered carbocyclic ring in which one or two carbon atoms are replaced by nitrogen.
  11. 11. The use of claim 10, wherein the tumors of the central and peripheral nervous system are selected from the group consisting of neuroblastoma, glioblastoma, and medulloblastoma.
HK15100468.6A 2007-07-20 2014-10-15 Substituted indazole derivatives active as kinase inhibitors HK1200162B (en)

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