HK1239665A1

HK1239665A1 - 6-hydroxybenzofuranyl- and 6-alkoxybenzofuranyl-substituted imidazopyridazines

Info

Publication number: HK1239665A1
Application number: HK17112902.3A
Authority: HK
Inventors: Knut Eis; Ingo Hartung; Kirstin Petersen; Philip Lienau; Ulf Bömer
Original assignee: 拜耳医药股份公司
Priority date: 2014-12-23
Filing date: 2015-12-21
Publication date: 2018-05-11

Description

6-hydroxybenzofuranyl-and 6-alkoxybenzofuranyl-substituted imidazopyridazines

Technical Field

The present invention relates to 6-hydroxybenzofuranyl-and 6-alkoxybenzofuranyl-substituted imidazopyridazine compounds of general formula (I) as described and defined herein, to methods of preparing said compounds, to intermediate compounds useful for preparing said compounds, to pharmaceutical compositions and combinations comprising said compounds and to the use of said compounds for manufacturing a pharmaceutical composition for the treatment or prophylaxis of a disease, in particular of a hyper-proliferative and/or angiogenesis disorder, as a sole agent or in combination with other active ingredients.

Background

The present invention relates to compounds that inhibit MKNK1 kinase (also known as MAP kinase interacting kinase, Mnk1) and MKNK2 kinase (also known as MAP kinase interacting kinase, Mnk 2). Human MKNK comprises a set of four proteins encoded by two genes (gene symbols: MKNK1 and MKNK2) obtained by alternative splicing. Type b lacks the C-terminal MAP kinase binding domain. The catalytic domains of MKNK1 and MKNK2 are very similar and contain a unique DFD (Asp-Phe-Asp) motif in subdomain VII, which is typically DFG (Asp-Phe-Gly) among other protein kinases and is believed to alter ATP binding [ Jauch et al, Structure 13, 1559-. MKNK1a binds to and is activated by ERK and p38MAP kinases, but not JNK 1. MKNK2a binds to and is only activated by ERK. MKNK1b had low activity under all conditions, while MKNK2b had basal activity independent of ERK or p38MAP kinase [ Buxade M et al, Frontiers in Bioscience 5359-5374, 1/5 of 2008 ].

MKNK phosphorylated eukaryotic initiation factor 4E (eIF4E), heterogeneous nuclear RNA-binding protein A1(hnRNPA1), polypyrimidine sequence-binding protein-related splicing factor (PSF), cytoplasmic phospholipase A2(cPLA2) and Sprouty 2(hSPRY2) [ Buxade M et al, Frontiers in Bioscience 5359-5374,2008, 5.1).

eIF4E is an oncogene that is amplified in many cancers and is phosphorylated only by MKNK protein, as shown in the KO-mouse study [ koniek et al, Cell Cycle 7:16,2466-2471, 2008; ueda et al, Mol Cell Biol 24, 6539-. eIF4E plays a key role in achieving translation of cellular mRNA. eIF4E binds to the 7-methylguanosine cap at the 5' end of cellular mRNA and delivers them to the ribosome as part of the eIF4F complex, which also includes eIF4G and eIF 4A. Although all capped mrnas require eIF4E for translation, mRNA pools were aberrantly dependent on elevated eIF4E activity for translation. These so-called "weak mRNAs" are often poorly translated due to their long and complex 5' UTR region, and they encode proteins that play an important role in all aspects of malignancy, including VEGF, FGF-2, c-Myc, cyclin D1, survivin, BCL-2, MCL-1, MMP-9, heparanase, and others. expression and function of eIF4E is elevated in a variety of human cancers and is directly associated with disease progression [ Konicek et al, Cell Cycle 7:16,2466-2471,2008 ].

MKNK1 and MKNK2 are the only kinases known to phosphorylate eIF4E at Ser 209. The overall translation rate is not affected by phosphorylation of eIF4E, but it has been suggested that eIF4E phosphorylation promotes polysome formation (i.e., multiple ribosomes on a single mRNA) that ultimately enables "weak mrnas" to be translated more efficiently [ Buxade M et al, Frontiers in bioscience 5359-5374, 1/5 2008 ]. Alternatively, phosphorylation of eIF4E by MKNK protein facilitates the release of eIF4E from the 5' cap, such that the 48S complex can move along the "weak mRNA", thereby locating the initiation codon [ Blagden SP and WillisaE, Nat Rev Clin Oncol.8(5):280-91,2011 ]. Thus, increased eIF4E phosphorylation predicts a poor prognosis for non-small cell lung Cancer patients [ Yoshizawa et al, Clin Cancer Res.16(1):240-8,2010 ]. Other data suggest a functional role for MKNK 1in carcinogenesis, as overexpression of constitutively activated MKNK1 (but not kinase-inactivated MKNK1) in mouse embryonic fibroblasts promoted tumor formation [ Chrestensen c.a. et al, Genes cells12, 1133-1140,2007 ]. Furthermore, increased phosphorylation and activity of MKNK protein in breast cancer is associated with overexpression of HER2 [ Chrestensen, c.a. et al, j.biol.chem.282, 4243-4252,2007 ]. In a model using E μ -Myc transgenic hematopoietic stem cells for tumor generation in mice, constitutive activation (rather than kinase inactivation) of MKNK1 also accelerated tumor growth. Comparable results were obtained when eIF4E with the S209D mutation was analyzed. The S209D mutation mimics phosphorylation at the MKNK1 phosphorylation site. In contrast, the non-phosphorylated form of eIF4E attenuated tumor growth [ Wendel HG et al, GenesDev.21(24):3232-7,2007 ]. Selective MKNK inhibitors that block phosphorylation of eIF4E induced apoptosis and inhibited proliferation of cancer cells and soft agar growth in vitro. This inhibitor also inhibited the growth halo of experimental B16 melanoma lung metastases and the growth of subcutaneous HCT116 colon cancer xenograft tumors without affecting body weight [ Konicek et al, cancer Res.71(5):1849-57,2011 ]. In summary, phosphorylation of eIF4E by MKNK protein activity promotes cell proliferation and survival and is critical for malignant transformation. Inhibition of MKNK activity may provide an easily manageable cancer treatment.

WO2007/025540a2(Bayer Schering Pharma AG) relates to substituted imidazo [1,2-b ] pyridazines as kinase inhibitors, in particular PKC (protein kinase C) inhibitors, in particular PKC θ inhibitors.

WO2007/025090a2(Kalypsis, Inc.) relates to heterocyclic compounds that are useful as inhibitors of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (Erk) kinase (abbreviated "MEK"). In particular, WO2007/025090a2 relates in particular to imidazo [1,2-b ] pyridazine.

WO2007/013673a1(Astellas Pharma Inc.) relates to fused heterocycles as inhibitors of lymphocyte protein tyrosine kinases (abbreviated as "LCK"). In particular, WO2007/013673a1 relates in particular to imidazo [1,2-b ] pyridazine.

WO2007/147646a1(Bayer Schering Pharma AG) relates to oxo-substituted imidazo [1,2-b ] pyridazines as kinase inhibitors, in particular PKC (protein kinase C) inhibitors, in particular PKC θ inhibitors.

WO2008/025822a1(cellzome (uk) Ltd.) relates to oxadiazodiazine derivatives for use as kinase inhibitors. In particular, WO2008/025822a1 relates to inter alia imidazo [1,2-b ] pyridazines for use as kinase inhibitors, in particular inducible T cell kinase (abbreviated "Itk") inhibitors.

WO2008/030579a2(Biogen Idec MA Inc.) relates to modulators of interleukin-1 (IL-1) receptor-associated kinase (abbreviated "IRAK"). In particular, WO2008/030579a2 relates in particular to imidazo [1,2-b ] pyridazine.

WO2008/058126a2(Supergen, Inc.) relates in particular to imidazo [1,2-b ] pyridazine derivatives as protein kinase inhibitors, in particular PIM kinase inhibitors.

WO2009/060197a1(Centro Nacional de investigations Oncology (CNIO)) relates to imidazopyridazines for use as protein kinase inhibitors, such as PIM family kinases.

US4,408,047(Merck & co., Inc.) relates in particular to imidazopyridazines with 3-amino-2-OR-propoxy substituents which have β -adrenergic blocking activity.

WO03/018020A1(Takeda Chemical Industries, Ltd.) relates to inhibitors against c-Jun N-terminal kinases comprising a compound which is in particular imidazo [1,2-b ] pyridazine.

WO2008/052734a1(Novartis AG) relates to heterocyclic compounds as anti-inflammatory agents. In particular, said compound is in particular imidazo [1,2-b ] pyridazine. The compounds are useful for treating diseases mediated by the ALK-5 and/or ALK-4 receptor, and also for treating diseases mediated by the PI3K receptor, the JAK-2 receptor, and the TRK receptor.

WO2008/072682a1(Daiichi Sankyo Company, Limited) relates to imidazo [1,2-b ] pyridazine derivatives having an inhibitory effect on TNF- α production, playing a role in pathological models of inflammatory and/or autoimmune diseases.

WO2008/079880a1(Alcon Research, Ltd.) relates to 6-aminoimidazo [1,2-b ] pyridazine analogs useful as Rho-kinase inhibitors for the treatment of glaucoma and ocular hypertension.

WO2009/091374a2(Amgen Inc.) relates to fused heterocyclic derivatives. The selected compounds are effective in preventing and treating diseases, such as hepatocyte growth factor ("HGF") diseases.

WO2013/013188a1(Tolero Pharmaceuticals, Inc.) relates to heterocyclic derivatives for the treatment of cancer, autoimmunity, inflammation and other Pim kinase related disorders.

An article entitled "Structural base of inhibition specificity of the Protooncogene Proviral Insertion Site in molecular MurinLeukemia Virus (PIM-1) Kinase" in J.Med.Chem.,2005,48,7604-7614 discloses inter alia imidazo [1,2-b ] pyridazines as inhibitor structures for use in the studies described therein.

An article named "Discovery of Mitogen-activated protein Kinase-Interacting Kinase1Inhibitors by a comparative fragmentation-organized viral Screening Approach" in J.Med.chem.,2010,53, 6618-.

An article entitled "Therapeutic inhibition of MAP kinase inhibition blocks and immunological inhibition 4 Ephosphatation and supressures output of experimental residues" in Cancer Res, 3/1.2011, 71,1849-1857 discloses in particular that the known antifungal agent Cercosporamide (Cercoporamide) is an MKNK1 inhibitor.

WO 2013/041634(Bayer Intelligent Property GmbH), WO 2013/034570(Bayer Intelligent Property GmbH), WO 2013/144189(Bayer Intelligent Property GmbH) and WO 2014/128093(Bayer Pharma Aktiengesellschaft) relate to 4-methoxy-1-benzofuran-2-yl-substituted imidazo [1,2-b ] pyridazines and 5-methoxy-1-benzofuran-2-yl-substituted imidazo [1,2-b ] pyridazines as MKNK1 inhibitors.

However, the above prior art does not describe specific 6-hydroxy-1-benzofuran-2-yl-and 6-alkoxy-1-benzofuran-2-yl-substituted imidazo [1,2-b ] pyridazine compounds of general formula (I) of the present invention as defined herein, i.e. the imidazo [1,2-b ] pyridazinyl moiety as described and defined herein and as hereinafter referred to as "compounds of the present invention", which carries:

-at the 3-position:

a group;

-at the 6-position:

a group;

wherein R1 and R2 are as defined in the claims, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, or their pharmacological activity is not described.

The compounds of the invention have been found to have unexpected and superior properties, which form the basis of the present invention.

In particular, it has surprisingly been found that said compounds of the invention effectively inhibit MKNK-1 kinase and are therefore useful for the treatment or prevention of diseases caused by or accompanied by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response, in particular wherein said uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is mediated by MKNK-1 kinase, such as haematological tumours, solid tumours and/or their metastases, such as leukaemia and myelodysplastic syndrome, malignant lymphomas, head and neck tumours including brain tumours and brain metastases, diseases in which there is an inappropriate cellular immune response or inappropriate cellular inflammatory response, and in particular wherein said uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is mediated by MKNK-1 kinase Breast, gastrointestinal, endocrine, breast and other gynaecological tumours including non-small cell and small cell lung tumours, urological tumours including renal, bladder and prostate tumours, skin tumours and sarcomas, and/or metastases thereof.

The above prior art does not show that the specific 6-hydroxybenzofuranyl-and 6-alkoxybenzofuranyl-substituted imidazopyridazine compounds of general formula (I) of the invention defined herein would be so active as inhibitors of MKNK-1 kinase.

Disclosure of Invention

According to a first aspect, the invention covers compounds of general formula (I):

wherein:

r1 represents C₂-C₆-alkyl-or C₃-C₆-cycloalkyl, which is substituted by a group a, and which is optionally substituted once, twice or three times, independently of each other, by a substituent selected from:

halogen atom, CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₆-cycloalkyl-, phenyl, optionally substituted one or more times independently of each other by R substituents; heteroaryl-, which is optionally substituted one or more times, independently of each other, with an R substituent; -C (═ O) NH₂A group, -C (═ O) N (h) R ', -C (═ O) N (R ') R ", -C (═ O) OH, OR-C (═ O) OR ';

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂an-OC (═ O) NHR 'or-OC (═ O) N (R') R "group,

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group in which one carbon atom is optionally replaced by another heteroatom-containing group selected from: NH, O, S (═ O) and S (═ O)₂And 5-to 7-membered nitrogen-containing heterocycloalkyl wherein one additional ring atom is optionally replaced by C (═ O);

-said azetidinyl and said 5-to 7-membered nitrogen-containing heterocycloalkyl optionally substituted once or twice independently of each other by a substituent selected from:

halogen atom, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, or C₃-C₄-a cycloalkyl group;

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₆-an alkyl group;

r represents a substituent selected from:

halogen atom, CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₆-cycloalkyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-C(＝O)OH、-C(＝O)OR’、-NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂A N (R ') R' group;

r' and R "independently of each other represent a substituent selected from:

C₁-C₆-alkyl-, C₃-C₆-cycloalkyl-, C₁-C₆-haloalkyl-, C₁-C₆-alkoxy-C₂-C₆-alkyl-, or C₁-C₆-haloalkoxy-C₂-C₆-an alkyl-group;

or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same.

Definition of

Unless otherwise indicated, components optionally substituted as described herein may be substituted one or more times independently of each other at any possible position. When any variable occurs more than one time in any constituent, each definition is independent. For example, each definition of R, R 'and R "is independent whenever R, R' and/or R" occurs more than one time in any compound of formula (I).

In the case where the component contains more than one moiety, e.g. C₁-C₆-alkoxy-C₂-C₆-alkyl-, the position of possible substituents may be at any suitable position of any of these moieties. The hyphen at the beginning or end of a component marks the point of attachment to the rest of the molecule. In the case of substituted rings, the substituents can be in any suitable position of the ring, also on the ring nitrogen atom, if appropriate.

The terms mentioned herein preferably have the following meanings:

the term "halogen atom", "halo-" or "halo-" is to be understood as meaning a fluorine, chlorine, bromine or iodine atom, preferably a fluorine, chlorine, bromine or iodine atom. According to one embodiment, the terms "halogen atom", "halo-" or "halo-" are to be understood as meaning a fluorine atom. According to one embodiment, the terms "halogen atom", "halo-" or "halo-" are to be understood as meaning a chlorine atom.

The term "C₁-C₆Alkyl "is understood to mean preferably a straight-chain or branched, saturated, monovalent hydrocarbon radical having 1,2, 3,4, 5 or 6 carbon atoms, for example methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutanylA group, 1-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 3-dimethylbutyl, or 1, 2-dimethylbutyl, or an isomer thereof. In particular, the radicals have 1,2, 3 or 4 carbon atoms ("C)₁-C₄Alkyl groups) such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, or2, 3 or 4 carbon atoms ("C)₂-C₄Alkyl radicals), such as ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, more particularly having 1,2 or 3 carbon atoms ("C)₁-C₃Alkyl groups) such as methyl, ethyl, n-propyl or isopropyl.

The term "C₁-C₆Haloalkyl "is understood as preferably meaning a straight-chain or branched saturated monovalent hydrocarbon radical (where the term" C "is used)₁-C₆Alkyl "as defined above) and wherein one or more hydrogen atoms are substituted with the same or different halogen atoms (i.e., one halogen atom is independent of another). In particular, the halogen atom is F. Said C is₁-C₆Haloalkyl is, for example, -CF₃、-CHF₂、-CH₂F、-CF₂CF₃Or CH₂CF₃。

The term "C₁-C₆Alkoxy "is to be understood as meaning preferably a straight-chain or branched saturated monovalent hydrocarbon radical of the formula-O-alkyl (where the term" alkyl "is as defined above), for example methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, pentoxy, isopentoxy or n-hexoxy, or isomers thereof.

The term "C₁-C₆Haloalkoxy "is understood to mean preferably a straight-chain or branched saturated monovalent C₁-C₆Alkoxy (as defined above) in which one or more hydrogen atoms are the same orDifferent halogen atoms. In particular, the halogen atom is F. Said C₁-C₆Haloalkoxy is, for example, -OCF₃、-OCHF₂、-OCH₂F、-OCF₂CF₃or-OCH₂CF₃。

The term "C₁-C₆alkoxy-C₂-C₆Alkyl "is understood to mean preferably a straight-chain or branched saturated monovalent C₂-C₆Alkyl-alkyl (as defined above) in which one or more hydrogen atoms are replaced by the same or different C₂-C₆Alkoxy (as defined above) is substituted, e.g. methoxyalkyl, ethoxyalkyl, propoxyalkyl, isopropoxyalkyl, butoxyalkyl, isobutoxyalkyl, tert-butoxyalkyl, sec-butoxyalkyl, pentoxyalkyl, isopentoxyalkyl, hexyloxyalkyl (wherein the term "C" is used)₂-C₆Alkyl "as defined above), or isomers thereof.

The term "C₁-C₆halogenoalkoxy-C₂-C₆Alkyl "is understood to mean preferably a straight-chain or branched saturated monovalent C₁-C₆alkoxy-C₂-C₆Alkyl (as defined above) in which one or more hydrogen atoms are replaced by the same or different halogen atoms. In particular, the halogen atom is F. Said C is₁-C₆halogenoalkoxy-C₂-C₆Alkyl is, for example, -CH₂CH₂OCF₃、-CH₂CH₂OCHF₂、-CH₂CH₂OCH₂F、-CH₂CH₂OCF₂CF₃or-CH₂CH₂OCH₂CF₃。

The term "C₃-C₆Cycloalkyl is understood to mean a saturated, monovalent, monocyclic hydrocarbon ring containing 3,4, 5 or 6 carbon atoms ("C)₃-C₆-cycloalkyl "), for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In particular, the radicals have 3 or 4 carbon atoms ("C₃-C₄-cycloalkyl "), for example cyclopropyl or cyclobutyl. The term "5-to 7-membered nitrogen-containing heterocycloalkyl" is understood to mean a saturated monovalent monocyclic hydrocarbon ring containing 4, 5 or 6 carbon atoms and a nitrogen-containing group selected from N and NH, wherein one carbon atom is optionally replaced by another heteroatom-containing group selected from: NH, O, S (═ O) and S (═ O)₂And in said nitrogen-containing heterocycloalkyl, one additional ring atom is optionally replaced by C (═ O); the nitrogen-containing heterocycloalkyl group can be attached to the remainder of the molecule via any carbon or nitrogen atom.

Specifically, without limitation, the nitrogen-containing heterocycloalkyl group may be a 5-membered ring such as pyrrolidinyl, imidazolidinyl, or pyrazolidinyl, or a 6-membered ring such as piperidinyl, morpholinyl, thiomorpholinyl, or piperazinyl, or a 7-membered ring such as azepanyl, diazepanyl, or oxazepanyl; the nitrogen-containing heterocycloalkyl group can be attached to the remainder of the molecule via any carbon or nitrogen atom.

The term "heteroaryl" is understood as preferably meaning a monovalent, monocyclic aromatic ring system having 5 or 6 ring atoms (a "5-to 6-membered heteroaryl" group) which contains at least one heteroatom, which may be identical or different, for example oxygen, nitrogen or sulfur. Specifically, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl and the like, or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like.

Typically, unless otherwise specified, heteroaryl or heteroarylene includes all possible isomeric forms thereof, e.g., positional isomers thereof. Thus, for certain illustrative, non-limiting examples, the term pyridyl or pyridinylidene includes pyridin-2-yl, pyridin-2-ylidene, pyridin-3-yl, pyridin-3-ylidene, pyridin-4-yl, and pyridin-4-ylidene; or the term thienyl or thienylene includes thien-2-yl, thien-3-yl, and thien-3-yl.

As used throughout this document, for example at "C₁-C₆Alkyl group "," C₁-C₆Haloalkyl "," C₁-C₆Alkoxy "or" C₁-C₆The term "C" as used in the context of the definition of haloalkoxy₁-C₆"is to be understood as meaning alkyl having from 1 to 6 carbon atoms in a defined number, i.e. 1,2, 3,4, 5 or 6 carbon atoms. It is also understood that the term "C" refers to₁-C₆"should be interpreted as including any subrange therein, e.g. C₁-C₆、C₂-C₅、C₃-C₄、C₁-C₂、C₁-C₃、C₁-C₄、C₁-C₅(ii) a In particular C₁-C₂、C₁-C₃、C₁-C₄、C₁-C₅、C₁-C₆(ii) a More particularly C₁-C₄(ii) a In "C₁-C₆Haloalkyl "or" C₁-C₆In the case of haloalkoxy ", even more particularly C₁-C₂。

Further, as used herein, throughout this document, for example at "C₃-C₆The term "C" as used in the context of the definition of-cycloalkyl₃-C₆"is understood to mean cycloalkyl having 3 to 6 defined number of carbon atoms, i.e. 3,4, 5 or 6 carbon atoms. It is also understood that the term "C" refers to₃-C₆"should be interpreted as including any subrange therein, e.g. C₃-C₆、C₄-C₅、C₃-C₅、C₃-C₄、C₄-C₆，C₅-C₆(ii) a In particular C₃-C₆。

The term "substituted" means that one or more hydrogens on the designated atom is replaced with an option from the designated group, provided that the designated atom's normal valency at the present time is not exceeded and that the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

The term "optionally substituted" means optionally substituted with a specified group, radical or moiety.

A substituent of a ring system refers to a substituent attached to an aromatic or non-aromatic ring system, e.g. replacing an available hydrogen on the ring system.

The term "one or more", as used herein, for example in the definition of a substituent of a compound of the general formula in accordance with the invention, is understood to mean "one, two, three, four or five, in particular one, two, three or four, more in particular one, two or three, even more in particular one or two".

The invention also includes all suitable isotopic variations of the compounds of the invention. Isotopic variations of the compounds of the present invention are defined as those in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually or predominantly found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine and iodine, such as²H (deuterium),³H (tritium),¹¹C、¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、³²P、³³P、³³S、³⁴S、³⁵S、³⁶S、¹⁸F、³⁶Cl、⁸²Br、¹²³I、¹²⁴I、¹²⁹I and¹³¹I. some isotopic variations of the compounds of the present invention, e.g. wherein one or more are incorporated, e.g.³H or¹⁴Those of the radioactive isotopes of C are useful for drug and/or substrate tissue distribution studies. Tritiated and carbon-14 (i.e., tritiated) are particularly preferred due to ease of preparation and detectability¹⁴C) An isotope. In addition, substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, e.g. increased in vivo half-lifeOr reduced dosage requirements, and is therefore preferred in some circumstances. Isotopic variations of the compounds of the present invention can generally be prepared by conventional methods known to those skilled in the art, for example by the exemplary procedures or preparative procedures described in the examples below, using appropriate isotopic variations of appropriate reagents.

When the plural form of the words compound, salt, polymorph, hydrate, solvate and the like are used herein, it is to be understood that reference to a compound, salt, polymorph, isomer, hydrate, solvate and the like in the singular is also intended.

"stable compound" or "stable structure" refers to a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture and formulation into an effective therapeutic agent.

The compounds of the present invention may contain one or more asymmetric centers, depending on the location and nature of the various substituents desired. Asymmetric carbon atoms may exist in either the (R) or (S) configuration, resulting in a racemic mixture in the case of one asymmetric center and a diastereomeric mixture in the case of multiple asymmetric centers. In some cases, asymmetry may also exist due to hindered rotation about a particular bond, for example, the central bond connecting two substituted aromatic rings of a particular compound.

The compounds of the invention may contain a sulfur atom, which may be asymmetric, such as an asymmetric sulfoxide group of the structure:

wherein denotes an atom which may be bonded to the rest of the molecule.

The ring substituents may also be present in cis or trans form. All such configurations (including enantiomers and diastereomers) are intended to be included within the scope of the present invention.

Preferred compounds are those that produce a more desirable biological activity. Isolated, purified or partially purified isomers and stereoisomers, or racemic or diastereomeric mixtures of the compounds of the invention are included within the scope of the invention. Purification and isolation of such materials can be accomplished by standard techniques known in the art.

Optical isomers may be obtained by resolution of the racemic mixture according to conventional methods, for example by formation of diastereomeric salts using optically active acids or bases, or by formation of covalent diastereomers. Examples of suitable acids are tartaric acid, diacetyltartaric acid, ditoluoyltartaric acid and camphorsulfonic acid. Mixtures of diastereomers may be separated into their individual diastereomers on the basis of their physical and/or chemical differences by methods known in the art, e.g., by chromatography or fractional crystallization. The optically active base or acid is then released from the separated diastereoisomeric salt. Another different method of separating optical isomers involves the use of chiral chromatography (e.g., a chiral HPLC column) with or without conventional derivatization, which can be optimally selected to maximize separation of the enantiomers. Suitable chiral HPLC columns are manufactured by Daicel, e.g., chiralel OD and chiralel OJ, all of which are routinely selected. Enzymatic separation may also be used with or without derivatization. Likewise, the optically active compounds of the present invention can be obtained by chiral synthesis using optically active starting materials.

To distinguish the different types of isomers from each other, reference is made to IUPAC Rules Section E (Pure Appl Chem 45,11-30,1976).

The present invention includes all possible stereoisomers of the compounds of the invention, which are single stereoisomers or any mixture of said stereoisomers (e.g. R-or S-isomers, or E-or Z-isomers) in any proportion. Separation of the individual stereoisomers (e.g. individual enantiomers or individual diastereomers) of the compounds of the invention may be achieved by any suitable prior art method, such as chromatography, particularly, for example, chiral chromatography.

In addition, the compounds of the present invention may exist in tautomeric forms. For example, any compound of the invention comprising a pyrazole moiety as heteroaryl may, for example, exist in the form of a 1H tautomer or a 2H tautomer, or even in the form of a mixture of any amounts of the two tautomers, or any compound of the invention comprising a triazole moiety as heteroaryl may, for example, exist in the form of a 1H tautomer, a 2H tautomer, or a 4H tautomer, or even in the form of a mixture of any amounts of the 1H, 2H, and 4H tautomers, i.e.:

the present invention includes all possible tautomers of the compounds of the invention, either as single tautomers or as any mixtures of said tautomers, in any ratio.

In addition, the compounds of the present invention may exist in the form of N-oxides, which are defined as compounds of the present invention in which at least one nitrogen is oxidized. The present invention includes all such possible N-oxides.

The invention also relates to useful forms of the compounds as disclosed herein, such as metabolites, hydrates, solvates, prodrugs, salts, especially pharmaceutically acceptable salts, and co-precipitates.

The compounds of the invention may be present in the form of hydrates or solvates, wherein the compounds of the invention comprise, for example, as structural element of the crystal lattice of the compound, a polar solvent, in particular water, methanol or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric proportions. In the case of stoichiometric solvates, such as hydrates, there may be semi (hemi-or hemi-) solvates or hemihydrate, mono-or monohydrate, sesquisolvates or sesquihydrates, di-or dihydrate, tri-or trihydrate, tetra-or tetrahydrate, penta-or pentahydrate, respectively, and the like. The present invention includes all such hydrates or solvates.

In addition, the compounds of the invention may exist in free form, e.g. as a free base, or as a free acid or zwitterion, or may exist in the form of a salt. The salt may be any salt, which may be an organic or inorganic addition salt, in particular any pharmaceutically acceptable organic or inorganic addition salt commonly used in pharmacy.

The term "pharmaceutically acceptable salts" refers to relatively non-toxic, inorganic or organic acid addition salts of the compounds of the present invention. See, for example, S.M.Berge et al, "Pharmaceutical Salts", J.pharm.Sci.1977,66, 1-19.

Suitable pharmaceutically acceptable salts of the compounds of the invention may be, for example, acid addition salts of the compounds of the invention which carry a nitrogen atom in the chain or ring and which are sufficiently basic, for example with the following inorganic acids: such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid (disufuric acid), phosphoric acid or nitric acid, or acid addition salts with organic acids such as: such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid (hemisulfuric acid), or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt such as a sodium or potassium salt, an alkaline earth metal salt such as a calcium or magnesium salt, an ammonium salt, or a salt with an organic base which affords a physiologically acceptable cation, for example a salt with: n-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1, 6-hexanediamine, ethanolamine, glucosamine, sarcosine, serinol, tris (hydroxymethyl) aminomethane, aminopropanediol, sovak-base, 1-amino-2, 3, 4-butanetriol. In addition, the basic nitrogen-containing groups may be quaternized with the following agents: lower alkyl halides such as methyl chloride, bromide and iodide, ethyl chloride, bromide and iodide, propyl chloride, bromide and iodide and butyl chloride, bromide and iodide; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate; long chain halides such as decyl chloride, bromide and iodide, lauryl chloride, bromide and iodide, myristyl chloride, bromide and iodide and stearyl chloride, bromide and iodide; aralkyl halides such as benzyl bromide and phenethyl bromide, and the like.

Those skilled in the art will also recognize that acid addition salts of the claimed compounds can be prepared by reacting the compounds with the appropriate inorganic or organic acid by any of a variety of known methods. Alternatively, the alkali metal salts and alkaline earth metal salts of the acidic compounds of the present invention are prepared by reacting the compounds of the present invention with an appropriate base by various known methods.

The present invention includes all possible salts of the compounds of the invention, which may be single salts or any mixture of said salts in any proportion.

In this context, in particular in the experimental section for the synthesis of intermediates and examples of the invention, when a compound is mentioned as a salt form with the corresponding base or acid, the exact stoichiometric composition of said salt form obtained by the corresponding preparation and/or purification method is in most cases unknown.

Unless otherwise specified, suffixes of chemical names or structural formulae such as "hydrochloride", "trifluoroacetate", "sodium salt" or "x HCl", "x CF₃COOH "," x Na + "should be understood as not being of stoichiometric specification, but only in salt form.

This similarly applies to the following cases: wherein synthetic intermediates or example compounds or salts thereof have been obtained as solvates, if defined, with unknown stoichiometric composition, by the described preparation and/or purification methods, as hydrates.

The term "in vivo hydrolysable ester" as used herein is understood to mean an in vivo hydrolysable ester of a compound of the invention which comprises a carboxy or hydroxy group, for example a pharmaceutically acceptable ester which can be hydrolysed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for the carboxyl group include, for example, alkyl esters, cycloalkyl esters and optionally substituted phenylalkyl esters, in particular benzyl esters, C₁-C₆Alkoxymethyl esters, e.g. methoxymethyl ester, C₁-C₆Alkanoyloxymethyl esters, e.g. pivaloyloxymethyl ester, phthalidyl ester, C₃-C₈cycloalkoxy-carbonyloxy-C₁-C₆Alkyl esters such as 1-cyclohexylcarbonyloxyethyl ester; 1, 3-dioxole-2-oxomethyl ester (1,3-dioxolen-2-onylmethyl ester), such as 5-methyl-1, 3-dioxole-2-oxomethyl ester; and C₁-C₆Alkoxycarbonyloxyethyl esters, such as 1-methoxycarbonyloxyethyl ester, and the esters may be formed on any of the carboxyl groups of the compounds of the invention.

In vivo hydrolysable esters of compounds of the invention which contain a hydroxy group include inorganic acid esters (e.g. phosphate esters) and [ alpha ] -acyloxyalkyl ethers and related compounds which are cleaved by in vivo hydrolysis of the ester to form the parent hydroxy group. Examples of [ α ] -acyloxyalkyl ethers include acetoxymethyl ether (acetoxymethyloxy) and 2, 2-dimethylpropionyloxymethyl ether (2, 2-dimethylpropionyloxymethyloxy). The selection of groups which form in vivo hydrolysable esters with hydroxyl groups include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, alkoxycarbonyl (to form alkyl carbonates), dialkyl carbamoyl and N- (dialkyl aminoethyl) -N-alkyl carbamoyl (to form carbamates), dialkyl aminoacetyl and carboxy acetyl groups. The present invention includes all such esters.

In addition, the present invention includes all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs, or mixtures of more than one polymorph in any ratio.

According to a second embodiment of the first aspect, the present invention covers compounds of the above general formula (I), wherein:

halogen atom, CN, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, C₃-C₄-cycloalkyl-, -C (═ O) NH₂A group, -C (═ O) N (h) R ', -C (═ O) N (R ') R ", -C (═ O) OH, OR-C (═ O) OR ';

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-、-OC(＝O)R’、-OC(＝O)NH₂-OC (═ O) NHR ', or-OC (═ O) N (R') R "groups,

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

r2 represents a substituent selected from:

hydrogen atom or C₁-C₃-an alkyl group;

r' and R "independently of each other represent a substituent selected from:

C₁-C₃-alkyl-, C₃-C₄-cycloalkyl-, C₁-C₃-haloalkyl-, C₁-C₃-alkoxy-C₂-C₃-alkyl-, or C₁-C₃-haloalkoxy-C₂-C₃-an alkyl-group;

According to a third embodiment of the first aspect, the present invention covers compounds of the above general formula (I), wherein:

r1 represents C₂-C₆-alkyl, substituted by a group a, and optionally substituted by C₁-C₃-the alkyl groups are substituted once, twice or three times independently of each other;

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group,

or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

-said 5-to 7-membered nitrogen-containing heterocycloalkyl optionally substituted by C₁-C₃-the alkyl groups are substituted once or twice independently of each other;

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₃-an alkyl group;

r 'and R' independently of one another represent C₁-C₃-an alkyl group;

According to a fourth embodiment of the first aspect, the present invention covers compounds of the above general formula (I), wherein:

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group,

or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group in which one carbon atom is optionally replaced by another heteroatom-containing group selected from: NH and O, and 5-to 7-membered nitrogen-containing heterocycloalkyl wherein one additional ring atom is optionally replaced by C (═ O);

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₃-an alkyl group;

r 'and R' independently of one another represent C₁-C₃-an alkyl group;

According to a fifth embodiment of the first aspect, the present invention covers compounds of the above general formula (I), wherein:

r1 represents C₂-C₄-an alkyl group, which is substituted by a group a, and which is optionally substituted once or twice by a methyl group;

wherein the group a represents a substituent selected from:

--NH₂-N (R ') R ', -N (H) C (═ O) R ', -OH, or methoxy,

or:

-a 5-or 6-membered nitrogen-containing heterocycloalkyl group,

-a 5-or 6-membered nitrogen-containing heterocycloalkyl group in which one carbon atom is optionally replaced by another heteroatom-containing group selected from: NH and O, and 5-or 6-membered nitrogen-containing heterocycloalkyl wherein one additional ring atom is optionally replaced by C (═ O);

-said 5-or 6-membered nitrogen-containing heterocycloalkyl optionally substituted with methyl;

r2 represents a substituent selected from:

a hydrogen atom, or a methyl group;

r 'and R' independently of one another represent methyl or ethyl;

In yet another embodiment of the above aspects, the present invention relates to a compound of formula (I), wherein:

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂-OC (═ O) NHR ', or-OC (═ O) N (R') R "groups,

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

halogen atom, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, or C₃-C₄-a cycloalkyl group.

halogen atom, CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₆-cycloalkyl-, phenyl, optionally substituted with ROne or more times independently of each other; heteroaryl-, which is optionally substituted one or more times, independently of each other, with an R substituent; -C (═ O) NH₂A group, -C (═ O) N (h) R ', -C (═ O) N (R ') R ", -C (═ O) OH, OR-C (═ O) OR ';

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂An — OC (═ O) NHR ', or — OC (═ O) N (R') R "group.

wherein the group a represents a substituent selected from:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、

-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂-OC (═ O) NHR ', or-OC (═ O) N (R') R "groups,

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、

-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂An — OC (═ O) NHR ', or — OC (═ O) N (R') R "group.

wherein the group a represents a substituent selected from:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group, or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

-said 5-to 7-membered nitrogen-containing heterocycloalkyl optionally substituted by C₁-C₃-alkyl groups are substituted once or twice independently of each other.

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group.

wherein the group a represents a substituent selected from:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group, or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

wherein the group a represents a substituent selected from:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

wherein the group a represents a substituent selected from:

--NH₂-N (R ') R ', -N (H) C (═ O) R ', -OH, or methoxy,

or:

-a 5-or 6-membered nitrogen-containing heterocycloalkyl group,

-said 5-or 6-membered nitrogen-containing heterocycloalkyl group, optionally substituted with a methyl group.

wherein the group a represents a substituent selected from:

--NH₂-N (R ') R ", -N (h) C (═ O) R', -OH, or methoxy.

wherein the group a represents a substituent selected from:

-a 5-or 6-membered nitrogen-containing heterocycloalkyl group,

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₆-an alkyl group.

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₃-an alkyl group.

r2 represents a substituent selected from:

a hydrogen atom, or a methyl group.

r2 represents a hydrogen atom.

r2 represents a methyl group.

r represents a substituent selected from:

halogen atom, CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₆-cycloalkyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-C(＝O)OH、-C(＝O)OR’、-NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂N (R ') R' group.

r' and R "independently of each other represent a substituent selected from:

C₁-C₆-alkyl-, C₃-C₆-cycloalkyl-, C₁-C₆-haloalkyl-, C₁-C₆-alkoxy-C₂-C₆-alkyl-, or C₁-C₆-haloalkoxy-C₂-C₆-an alkyl-group.

r' and R "independently of each other represent a substituent selected from:

C₁-C₃-alkyl-, C₃-C₄-cycloalkyl-, C₁-C₃-haloalkyl-, C₁-C₃-alkoxy-C₂-C₃-alkyl-, or C₁-C₃-haloalkoxy-C₂-C₃-an alkyl-group.

r 'and R' independently of one another represent C₁-C₃-an alkyl group.

r 'and R' independently of one another represent a methyl or ethyl group.

In a further embodiment of the above aspects, the present invention relates to a compound of formula (I), according to any of the above embodiments, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same.

In a further embodiment of the above aspect, the present invention relates to a compound of formula (I) in isolated form, in particular a compound of general formula (I) in isolated form as disclosed in the examples section below.

It will be appreciated that the present invention relates to any subcombination within any embodiment or aspect of the invention of the compounds of formula (I) above.

More specifically, the present invention encompasses the compounds of general formula (I) disclosed in the examples section below.

According to another aspect, the present invention encompasses a method of preparing a compound of the invention, said method comprising the steps described in the experimental section herein.

According to a further aspect, the present invention covers intermediate compounds which are useful in the preparation of the compounds of the invention of general formula (I), especially in the processes described herein. In particular, the invention encompasses compounds of general formula (E):

wherein R2 is as defined above for the compound of general formula (I), wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group. According to a further aspect, the present invention covers intermediate compounds which are useful in the preparation of the compounds of the invention of general formula (I), especially in the processes described herein. In particular, the invention encompasses compounds of general formula (V):

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R7 represents a protecting group, for example a silyl protecting group, for example a tert-butyldimethylsilyl group.

According to a further aspect, the present invention covers intermediate compounds which are useful in the preparation of the compounds of the invention of general formula (I), especially in the processes described herein. In particular, the invention encompasses compounds of general formula (W):

wherein R1 is as defined above for the compound of general formula (I), and wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl.

According to a further aspect, the present invention covers intermediate compounds which are useful in the preparation of the compounds of the invention of general formula (I), especially in the processes described herein. In particular, the invention encompasses compounds of general formula (X):

wherein X represents a leaving group, e.g. a halogen atom, e.g. a chlorine, bromine or iodine atom, or perfluoroAlkyl sulfonate ester groups, e.g. trifluoromethylsulfonate ester group, nonafluorobutylsulfonate ester group, and wherein R8 represents C₁-C₆An alkyl group.

According to a further aspect, the present invention covers the use of intermediate compounds of general formula (D) for the preparation of compounds of general formula (I) as defined above,

wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group. According to a further aspect, the present invention covers the use of intermediate compounds of general formula (E) for the preparation of compounds of general formula (I) as defined above,

wherein R2 is as defined above for the compound of general formula (I), wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

According to a further aspect, the present invention covers the use of an intermediate compound of formula (E') for the preparation of a compound of formula (I) as defined above,

wherein R1 is as defined above for the compound of formula (I), and wherein Y represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

According to a further aspect, the present invention covers the use of intermediate compounds of formula (V) for the preparation of compounds of formula (I) as defined above,

According to a further aspect, the present invention covers the use of intermediate compounds of general formula (W) for the preparation of compounds of general formula (I) as defined above,

According to a further aspect, the present invention covers the use of intermediate compounds of general formula (X) for the preparation of compounds of general formula (I) as defined above,

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R8 represents C₁-C₆An alkyl group.

Yet another aspect of the invention are intermediates for the synthesis of compounds of formula (I) as described herein and their use for the synthesis of compounds of formula (I). Preferred intermediates are the intermediate examples disclosed below.

Experimental part

The following table lists the abbreviations used in this paragraph and in the examples section.

Synthesis of compounds (overview):

the compounds of the invention can be prepared as described in the section below. Schemes 1 to 8 and the following procedures illustrate general synthetic routes for the compounds of general formula (I) of the present invention and are not intended to be limiting. It is clear to those skilled in the art that the conversion order illustrated in schemes 1 to 8 can be changed in various ways. Therefore, the conversion order illustrated in schemes 1 to 8 is not limiting. Further, interconversion of any substituents (R1 and R2) may be achieved before and/or after the exemplified transformations. These modifications may be, for example, the introduction of protecting groups; cleavage of the protecting group; exchange of functional groups; reduction or oxidation; halogenation; metallization, substitution, or other reactions known to those skilled in the art. These transformations include transformations that introduce functional groups that allow further interconversion of substituents. Suitable protecting Groups and their introduction and cleavage are well known to those skilled in the art (see, e.g., p.g.m.wuts and t.w.greene in "Protective Groups in Organic Synthesis", 4th edition, Wiley 2006). Specific examples are described in subsequent paragraphs. Furthermore, it is possible that two or more consecutive steps may be carried out without carrying out work-up between said steps, for example a "one-pot" reaction as known to the person skilled in the art.

Scheme 1:

wherein R1 and R2 are as defined above for the compound of formula (I), and wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

In a first step, a compound of formula A bearing a suitable X substituent (i.e., dichloropyridazine) may be reacted with ammonia at elevated temperature and pressure to provide a compound of formula B. [ similar to WO200733080, the entire contents of which are incorporated herein by reference ]

In a second step, the compound of the general formula B is reacted with, for example, chloroacetaldehyde diacetal (chloroacetaldehyde diacetal) or bromoacetaldehyde diacetal (bromoaldehyde diacetal) to give a bicyclic ring system C [ analogously to DE102006029447, the entire content of which is incorporated herein by reference ].

Activation of the 3-position of the bicyclic system to give compounds of general formula D can be accomplished, for example, by: the compound of formula C is brominated or iodinated using N-bromo-succinimide or N-iodo-succinimide, respectively.

In a fourth step, the introduction of the benzofuranyl residue can be effected using a suitably catalyzed cross-coupling reaction, using, for example, boric acid or stannane, which leads to a compound of the general formula E.

The compound of formula E acts as a central intermediate for the introduction of various side chains containing an alcohol function, which leads to the imidazopyridazinyl ether of formula (I). Introduction of the side chain can be achieved, for example, by using a base such as sodium hydride. Depending on the nature of the side chains, it may be necessary to carry out these reactions at elevated temperatures. It may also be desirable to introduce side chains modified with suitable protecting groups on functional groups that may interfere with the desired reaction.

The fourth and fifth steps of the sequence may also be switched to each other as shown in scheme 2.

Scheme 2:

Building blocks suitable for the introduction of benzofuranyl moieties may be prepared, for example, as shown in schemes 3a and 3 b.

Scheme 3 a:

wherein R7 represents a protecting group such as a silyl protecting group, e.g. tert-butyldimethylsilyl, and R9 represents a boronic acid-B (OH)₂Or a borate ester, and R10 represents a stannyl group, such as a tri-n-butylstannyl group.

Starting from hydroxybenzofuran F, the — OH function can be protected by a silylation method (e.g. using tert-butyldimethylsilyl chloride in the presence of imidazole as base), for example as a silyl ether, which leads to compounds of general formula G. After introduction of the R7 moiety, the 2-position of benzofuran can be activated to give compounds of general formula H after deprotonation with a strong base such as butyl lithium and reaction with a trialkylborate such as triisopropyl borate or with bis (pinacolato) diborane [ see, e.g., WO2009154780 or ACSMedicinal Chemistry Letters,2011, volume 2, page 97 ] to give compounds of general formula H.

Alternatively, after deprotonation with a strong base such as butyl lithium, the compound of formula G can be reacted with a trialkyltin halide such as tributyltin chloride [ see, for example, Bioorganic & Medicinal Chemistry,2012, vol.20, p.2762-2772 ], to give the corresponding stannyl benzofuran of formula J, which is also suitable for cross-coupling reactions as used in schemes 1 or 2.

Alternatively, building blocks suitable for the introduction of benzofuranyl moieties may be prepared, for example, as shown in scheme 3 b.

Scheme 3 b:

wherein R8 represents C₁-C₆-alkyl, and R9 represents boronic acid-B (OH)₂Or a borate ester, and R10 represents a stannyl group, such as a tri-n-butylstannyl group.

Starting from the hydroxybenzofuran F, an alkyl residue R8 can be introduced (e.g. using standard alkylation methods), which leads to a compound of general formula L. After introduction of the R8 moiety, the 2-position of the benzofuran can be activated to give compounds of general formula M after deprotonation with a strong base such as butyl lithium and reaction with a trialkylborate such as triisopropyl borate or with bis (pinacolato) diborane [ see, e.g., WO2009154780 or ACS Medicinal Chemistry Letters,2011, volume 2, page 97 ] to perform a cross-coupling reaction as shown in scheme 1 or 2.

Alternatively, after deprotonation with a strong base such as butyl lithium, the compound of formula L can be reacted with a trialkyltin halide such as tributyltin chloride [ see, for example, Bioorganic & Medicinal Chemistry,2012, Vol.20, p.2762-2772 ], to give the corresponding stannyl benzofuran of formula N, which is also suitable for cross-coupling reactions as used in schemes 1 or 2.

Scheme 4 shows a route for the synthesis of compounds of general formula (I-a), which are compounds of general formula (I) wherein-OR 2 represents hydroxy.

Scheme 4:

wherein R1 is as defined above for compounds of general formula (I), and wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, and wherein R10 represents a stannyl group, such as a tri-n-butylstannyl group, and wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example, a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

The compounds of formula (I-a) can be prepared according to the procedure described in scheme 1 using a coupling reaction of a compound of formula D as described in scheme 3a with a boronic acid or boronic ester of formula H or with a stannyl benzofuran of formula J to give a compound of formula (V), which is then reacted with an alcohol of formula K to introduce a residue R1 to give a compound of formula (W), followed by cleavage of the protecting group to give the compound of formula (I-a).

Scheme 5 illustrates an alternative synthesis of compounds of general formula (W).

Scheme 5:

wherein R1 is as defined above for compounds of general formula (I), and wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, and wherein R10 represents a stannyl group, such as a tri-n-butylstannyl group, and wherein Y represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

Alternatively, compounds of formula (W) may be prepared according to the method described in scheme 2 using a coupling reaction of a compound of formula E' as described in scheme 3a with a boronic acid or boronic ester of formula H or with a stannyl benzofuran of formula J.

Scheme 6 illustrates a route for the synthesis of compounds of general formula (I-b), which are compounds of general formula (I) wherein-OR 2 represents C₁-C₆An alkoxy group.

Scheme 6:

wherein R1 is as defined above for compounds of general formula (I), and wherein R8 represents C₁-C₆Alkyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, and wherein R10 represents a stannyl group, such as a tri-n-butylstannyl group, and wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example, a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

Compounds of formula (I-b) can be prepared according to the procedure described in scheme 1 using a coupling reaction of a compound of formula D as described in scheme 3b with a boronic acid or boronic ester of formula M or with a stannyl benzofuran of formula N to give a compound of formula (X), which is then reacted with an alcohol of formula K to introduce a residue R1 to give a compound of formula (I-b).

Scheme 7 illustrates an alternative synthesis of compounds of general formula (I-b), which is a compound of general formula (I) wherein-OR 2 represents C₁-C₆An alkoxy group.

Scheme 7:

wherein R1 is as defined above for compounds of general formula (I), and wherein R8 represents C₁-C₆Alkyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, and wherein R10 represents a stannyl group, such as a tri-n-butylstannyl group, and wherein Y represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group.

Alternatively, compounds of formula (I-b) may be prepared according to the method described in scheme 2 using a coupling reaction of a compound of formula E' as described in scheme 3b with a boronic acid or boronic ester of formula M or with a stannyl benzofuran of formula N.

Scheme 8 illustrates an alternative synthesis of compounds of general formula (I-b), which is a compound of general formula (I) wherein-OR 2 represents C₁-C₆An alkoxy group.

Scheme 8

Wherein R1 is as defined above for compounds of general formula (I), and wherein R8 represents C₁-C₆Alkyl-or C₁-C₆A haloalkyl group.

The compound of formula (I-a) may be converted to the compound of formula (I-b) using an alkylation process, for example, by reaction with an alkyl halide in the presence of a suitable base such as potassium carbonate.

Synthesis of the Compounds of the general formula (I) according to the invention

Compounds of general formula (I) wherein R1 and R2 have the meanings as given for general formula (I) can be synthesized according to the procedures shown in schemes 1 and 2. These schemes illustrate a main route that allows R1 to change at different synthesis stages. However, other routes may be used to synthesize the target compound, according to common general knowledge of those skilled in the art of organic synthesis.

The compounds of general formula (I-a) are compounds of general formula (I) wherein R1 has the meaning as given for general formula (I) and wherein-OR 2 represents hydroxy, which can be synthesized according to the procedures shown in schemes 1,2, 3a, 4 and 5. These schemes illustrate a main route that allows R1 to change at different synthesis stages. However, other routes may be used to synthesize the target compound, according to common general knowledge of those skilled in the art of organic synthesis.

The compounds of the general formula (I-b) are compounds of the general formula (I) in which R1 has the meaning as given for the general formula (I) and in which-OR 2 represents C₁-C₆-alkoxy groups, which can be synthesized according to the procedures shown in schemes 1,2, 3b, 6, 7 and 8. These schemes illustrate a main route that allows R1 to change at different synthesis stages. However, other routes may be used to synthesize the target compound, according to common general knowledge of those skilled in the art of organic synthesis.

According to one embodiment, the present invention also relates to a process for the preparation of a compound of the above general formula (I-a), said process comprising the steps of:

reacting the intermediate compound of formula (D) with a compound of formula (H) to thereby obtain a compound of formula (V),

the general formula (D) is:

wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group,

the general formula (H) is:

wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, or a salt of a carboxylic acid,

the general formula (V) is:

1. reacting the intermediate compound of formula (V) with a compound of formula (K) to thereby obtain a compound of formula (W),

the general formula (V) is:

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R7 represents a protecting group, such as a silyl protecting group, such as a tert-butyldimethylsilyl group,

the general formula (K) is:

R1-OH

(K)

wherein R1 is as defined above for the compound of formula (I),

the general formula (W) is:

wherein R1 is as defined above for compounds of general formula (I), and wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl, and

2. reacting a compound of formula (W) with tetra-n-butylammonium fluoride, thereby obtaining a compound of formula (I-a):

wherein R1 is as defined above for the compound of formula (I).

According to another embodiment, the present invention also relates to a process for the preparation of the compound of the above general formula (I-b), said process comprising the steps of:

1. reacting the intermediate compound of formula (E') with a compound of formula (H) to thereby obtain a compound of formula (W),

said general formula (E') is:

wherein R1 is as defined above for the compound of formula (I), and wherein Y represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group,

the general formula (H) is:

the general formula (W) is:

wherein R1 is as defined above for the compound of formula (I).

reacting the intermediate compound of formula (D) with a compound of formula (M) to thereby obtain a compound of formula (X),

the formula (D) is:

the general formula (M) is:

wherein R8 represents C₁-C₆Alkyl, and wherein R9 represents boronic acid-B (OH)₂Or a borate ester, or a salt of a carboxylic acid,

the general formula (X) is:

reacting the intermediate compound of formula (X) with a compound of formula (K) to thereby obtain a compound of formula (I-b),

the general formula (X) is:

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R8 represents C₁-C₆An alkyl group, a carboxyl group,

the general formula (K) is:

R1-OH

(K)

wherein R1 is as defined above for the compound of formula (I),

the general formula (I-b) is:

wherein R1 is as defined above for compounds of general formula (I), and wherein R8 represents C₁-C₆An alkyl group.

reacting the intermediate compound of formula (E') with a compound of formula (M) to thereby obtain a compound of formula (I-b),

said general formula (E') is:

the general formula (M) is:

wherein R8 represents C₁-C₆Alkyl radicalAnd wherein R9 represents boric acid-B (OH)₂Or a borate ester, or a salt of a carboxylic acid,

the general formula (I-b) is:

General part

The ACD/Name Batch version 12.02 was used to generate the chemical Name.

All reagents not described for synthesis in the experimental part are commercially available or synthesized as described in the literature.

HPLC method:

the method comprises the following steps:

the instrument comprises the following steps: waters Acquity UPLCMS ZQ 4000; column: acquity UPLC BEH C181.7 μm,50 × 2.1mm; eluent A: water +0.05 vol% formic acid, eluent B: acetonitrile +0.05 vol% formic acid, gradient: 1-99% B at 0-1.6min, 99% B at 1.6-2.0 min; the flow rate is 0.8 mL/min; temperature: 60 ℃; and (3) injection: 2 mu L of the solution; DAD scan: 210-400 nm; ELSD

The method 2 comprises the following steps:

the instrument comprises the following steps: waters Acquity UPLC-MS SQD; column: acquity UPLC BEH C181.750x2.1mm; eluent A: water +0.1 vol% formic acid (99%), eluent B: acetonitrile; gradient: 0-1.6min 1-99% B,1.6-2.0min 99% B; the flow rate is 0.8 mL/min; temperature: 60 ℃; and (3) injection: 2 mu L of the solution; DAD scan: 210-400 nm; ELSD.

Intermediates

Intermediate 1

3-bromo-6-chloroimidazo [1,2-b ] pyridazine

3-bromo-6-chloro-imidazo [1,2-b ] pyridazine is synthesized as described, for example, in WO2007/147646 or DE102006029447, for example, as follows:

step 1: preparation of 6-chloroimidazo [1,2-b ] pyridazine:

5.0g (38.6mmol) of 3-amino-6-chloropyridazine are heated in 15mL of n-butanol at 120 ℃ for 5 days together with 4.7mL (40mmol) of chloroacetaldehyde (55% strength in water). After completion of the reaction, the reaction mixture was added to a saturated sodium bicarbonate solution and extracted three times with ethyl acetate. The combined organic phases are then washed with saturated sodium chloride solution and dried over sodium sulfate, and the solvent is removed in vacuo. In the final purification by chromatography on silica gel, 4.17g (70%) of the desired product were isolated as an amorphous white solid.

¹H-NMR(CDCl₃Stored on molecular sieves): [ ppm of]＝7.06(1H)；7.79(1H)；7.92,(1H)；7.96(1H)ppm。

Step 2: preparation of 3-bromo-6-chloroimidazo [1,2-b ] pyridazine

478mg (3.11mmol) of 6-chloroimidazo [1,2-b ] pyridazine is introduced under argon into 10mL of chloroform, and 664mg (3.73mmol) of N-bromosuccinimide is added while cooling in ice. After the addition was complete, the reaction mixture was stirred at room temperature overnight. The reaction mixture was then mixed with water and ethyl acetate, and the phases were separated after addition of saturated sodium bicarbonate solution. The aqueous phase was extracted three more times with ethyl acetate. The combined organic phases are then washed with saturated sodium chloride solution and dried over sodium sulfate. In the final removal of the solvent in vacuo, the desired product was isolated in quantitative yield as an amorphous white solid, which was used in the subsequent reaction without further chromatographic purification.

¹H-NMR(CDCl₃Stored on molecular sieves): [ ppm of]＝7.12(1H)；7.79(1H)；7.90,(1H)ppm。

Intermediate 2

6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine

1.68g (7.22mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine (intermediate 1) was suspended in 124mL of 1, 4-dioxane. 1.46g (7.58mmol) of (6-methoxy-1-benzofuran-2-yl) boronic acid, 334mg (0.289mmol) of tetrakis (triphenylphosphino) palladium- (0) and 11mL of 2M aqueous sodium carbonate solution are added. The resulting mixture was heated to 105 ℃ and held for 16 hours.

The mixture was poured into a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The combined organic layers were washed with brine and dried over magnesium sulfate. After evaporation of the solvent, the resulting solid material was dissolved in dichloromethane, filtered and dried in vacuo to give 1.14g (53%) of the title compound as a solid material.

¹H-NMR(300MHz,DMSO-d₆)：[ppm]＝8.34(d,1H),8.28(s,1H),7.63(d,1H),7.51(s,1H),7.48(d,1H),7.25(d,1H),6.90(dd,1H),3.80(s,3H)。

LCMS (method 1): r_t＝1.30min；MS(ESIpos)m/z＝300[M+H]⁺。

Intermediate 3

6- (3- { [ tert-butyl (dimethyl) silyl ] oxy } propoxy) -3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine

16.4mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 1.6mL of anhydrous THF in an ice bath. 90.7mg (0.467mmol) of 3- { [ tert-butyl (dimethyl) silyl ] oxy } propan-1-ol were slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70.0mg (0.234mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) is added, the ice bath is removed and the resulting mixture is stirred at room temperature for 16 h.

The reaction mixture was carefully poured into saturated aqueous ammonium chloride solution. The aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated.

The crude product precipitated upon concentration was used in the subsequent step without further purification.

LC-MS (method 1): r_t＝1.75min；MS(ESIpos)m/z＝454[M+H]⁺。

Intermediate 4

(2R) -2- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-1-amine

In an ice bath, 1.2g (30mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 200mL of anhydrous THF. 2.26g (30mmol) of (2R) -1-aminopropan-2-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 5.0g (21.5mmol) 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine (intermediate 1) was added, the ice bath was removed and the resulting mixture was stirred at 60 ℃ for 16 h.

The reaction mixture was carefully poured into brine. The aqueous layer was extracted with dichloromethane. The combined organic layers were dried over magnesium sulfate and concentrated.

The crude product was purified by flash chromatography to yield 3.3g of the title compound.

LC-MS (method 1): r_t＝0.54min；MS(ESIpos)m/z＝272[M+H]⁺。

Intermediate 5

(1-benzofuran-6-yloxy) (tert-butyl) dimethylsilane

To 5g (37.3mmol) of 1-benzofuran-6-ol in 60mL of DMF was added 3.3g (48.5mmol) of imidazole. The mixture was cooled to 0 ℃ and 6.7g (44.7mmol) tert-butyl (chloro) -dimethylsilane was added. The mixture was stirred for 16 hours.

Brine was added, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated. The crude product was purified by flash chromatography to yield 6.3g of the title compound.

LC-MS (method 1): r_t＝1.68min；MS(ESIpos)m/z＝249[M+H]⁺。

Intermediate 6

(6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) boronic acid

To 3.0g (12.1mmol) of (1-benzofuran-6-yloxy) (tert-butyl) dimethylsilane (intermediate 5) in 51mL of THF was slowly added 7.25mL (18.1mmol) of a 2.5M solution of n-butyllithium in hexane. The resulting mixture was stirred at-78C for 90 minutes. 4.17mL (18.1mmol) of triisopropyl borate were added, and the mixture was stirred at room temperature for 16 hours.

10mL of 2M hydrochloric acid was added and stirring was continued at room temperature for 30 minutes.

The mixture was concentrated. Toluene was added and the mixture was concentrated again. This procedure was repeated with toluene and acetone to give 6.65g of the title compound as crude product, which was used in the subsequent step without further purification.

Intermediate 7

(2R) -2- { [3- (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine

1.6g (6.0mmol) of (2R) -2- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-1-amine (intermediate 4), 3.5g (12.1mmol) of (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) boronic acid (intermediate 6), 1.4g (1.21 μmol) of tetrakis (triphenylphosphine) palladium (0), and 9mL (18.1mmol) of potassium carbonate (c ═ 2mol/L in water) in 65mL of 1, 4-dioxane were heated to reflux for 16 h.

Saturated aqueous ammonium chloride and ethyl acetate were added successively. The organic layer was separated, dried over magnesium sulfate and evaporated.

The crude material was purified by flash chromatography to give 1.1g of crude product, which was used in the next step without further purification.

LC-MS (method 1): r_t＝1.21min；MS(ESIpos)m/z＝439[M+H]⁺。

Intermediate 8

N- { (2R) -2- [ (3-Bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propyl } acetamide

To 1.6g (5.98mmol) of N- { (2R) -2- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propyl } acetamide (intermediate 4) in 100mL of dichloromethane were added 1.9mL (23.9mmol) of pyridine and 1.13mL (12mmol) of acetic anhydride. The mixture was stirred at room temperature for 10 minutes.

The mixture was concentrated and the precipitate was purified by flash chromatography to give 1.08g of the title compound, which was used without further purification.

LC-MS (method 1): r_t＝0.75min；MS(ESIpos)m/z＝314[M+H]⁺。

Intermediate 9

N- [ (2R) -2- { [3- (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide

1.7g (5.4mmol) of N- { (2R) -2- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propyl } acetamide (intermediate 8), 3.2g (10.9mmol) (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) boronic acid (intermediate 6), 1.26g (1.09 μmol) of tetrakis (triphenylphosphine) palladium (0), and 8.1mL (16.3mmol) of potassium carbonate (c ═ 2mol/L in water) in 58mL 1, 4-dioxane were heated to reflux for 16 h.

The crude material was purified by flash chromatography to give 2g of crude product, which was used in the next step without further purification.

LC-MS (method 1): r_t＝1.54min；MS(ESIpos)m/z＝481[M+H]⁺。

Examples

Example 1

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-ol

To 131mg (0.289mmol) of 6- (3- { [ tert-butyl (dimethyl) silyl ] oxy } propoxy) -3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 3) in 4mL of THF is added 0.866mL (0.866mmol) of a 1M solution of tetra-n-butylammonium fluoride in THF. The mixture was stirred at room temperature for 16 hours.

The crude mixture was concentrated. The precipitate was taken up in DMF and purified by HPLC to give 42mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.10(s,1H),7.84(d,1H),7.50(d,1H),7.41(s,1H),7.05(d,1H),6.90(dd,1H),6.71(d,1H),4.64(t,2H),3.94(t,2H),3.88(s,3H),2.19(quin,2H)。

LC-MS (method 1): r_t＝1.02min；MS(ESIpos)m/z＝340[M+H]⁺。

Example 2

3- (6-methoxy-1-benzofuran-2-yl) -6- (3-methoxypropoxy) imidazo [1,2-b ] pyridazine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 43mg (0.47mmol) of 3-methoxypropan-1-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.234mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 72 h.

The reaction mixture was carefully poured into brine. The aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated.

The crude product was purified by HPLC to yield 27mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.12(s,1H),7.88(d,1H),7.54-7.45(m,2H),7.10(d,1H),6.91(dd,1H),6.77(d,1H),4.61(t,2H),3.89(s,3H),3.63(t,2H),3.41(s,3H),2.20(quin,2H)。

Example 3

2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } ethanamine

14mg (0.35mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 1.6mL of anhydrous THF in an ice bath. 25mg (0.40mmol) of 2-aminoethanol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 60mg (0.2mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 16 h.

The reaction mixture was carefully poured into brine. The aqueous layer was extracted with a 9:1 mixture of ethyl acetate and methanol. The combined organic layers were dried over magnesium sulfate and concentrated.

The crude product was purified by HPLC to give 48mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆)：[ppm]＝8.33(s,1H),8.12(d,1H),8.06(s,1H),7.56(d,1H),7.52(s,1H),7.23(d,1H),6.99(d,1H),6.89(dd,1H),4.60(t,2H),3.78(s,3H),3.23(t,2H)。

LC-MS (method 1): r_t＝0.72min；MS(ESIpos)m/z＝325[M+H]⁺。

Example 4

2- (6- { [ (2R) -1-Aminoprop-2-yl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) -1-benzofuran-6-ol

To 310mg (0.71mmol) of (2R) -2- { [3- (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine (intermediate 7) in 10mL of THF are added 446mg (1.4mmol) of tetra-n-butylammonium fluoride trihydrate. The mixture was stirred at room temperature for 10 minutes.

Brine was added and the crude mixture was concentrated. The precipitate was purified by HPLC to give 127mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆)：[ppm]＝8.27(s,1H),8.17(d,1H),8.10-8.08(m,1H),7.54-7.50(m,1H),7.47(d,1H),7.00(d,1H),6.97(d,1H),6.80(dd,1H),5.34-5.25(m,1H),3.03(d,2H),1.48(d,3H)。

LC-MS (method 1): r_t＝0.62min；MS(ESIpos)m/z＝325[M+H]⁺。

Example 5

N- [ (2R) -2- { [3- (6-hydroxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide

To 1.97g (4.1mmol) of N- [ (2R) -2- { [3- (6- { [ tert-butyl (dimethyl) silyl ] oxy } -1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide (intermediate 8) in 100mL of THF are added 2.59mg (8.2mmol) of tetra-N-butylammonium fluoride trihydrate. The mixture was stirred at room temperature for 24 hours.

Brine was added. The mixture was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The crude material obtained was purified by flash chromatography. The material from flash chromatography was dissolved in a mixture of dichloromethane and hexane to give 950mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆)：[ppm]＝9.67(s,1H),8.16-8.05(m,3H),7.51(t,2H),6.98(d,1H),6.92(d,1H),6.78(dd,1H),5.40-5.28(m,1H),3.54-3.40(m,2H),1.80(s,3H),1.42(d,3H)。

LC-MS (method 1): r_t＝0.81min；MS(ESIpos)m/z＝367[M+H]⁺。

Example 6

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine

14mg (0.35mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2.7mL of anhydrous THF in an ice bath. 31mg (0.40mmol) of 3-aminopropanol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 60mg (0.2mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 16 h.

The crude product was purified by HPLC to yield 43mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆)：[ppm]＝8.37(s,1H),8.09(d,1H),8.04(s,1H),7.59-7.53(m,2H),7.22(d,1H),6.98(d,1H),6.89(dd,1H),4.53(t,2H),3.78(s,3H),2.97(t,2H),2.10(quin,2H)。

LC-MS (method 1): r_t＝0.75min；MS(ESIpos)m/z＝339[M+H]⁺。

Example 7

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -N, N,2, 2-tetramethylpropan-1-amine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 1.6mL of anhydrous THF in an ice bath. 63mg (0.467mmol) of 3- (dimethylamino) -2, 2-dimethylpropan-1-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 22 h.

The crude product was dissolved in a mixture of methanol and DMF to give 53mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆)：[ppm]＝8.11(d,1H),8.05(s,1H),7.57-7.46(m,2H),7.25(s,1H),7.00(d,1H),6.89(dd,1H),4.24(s,2H),3.80(s,3H),2.30-2.21(m,8H),1.00(s,6H)。

LC-MS (method 1): r_t＝0.86min；MS(ESIpos)m/z＝395[M+H]⁺。

Example 8

4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } butan-1-amine

14mg (0.35mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2.7mL of anhydrous THF in an ice bath. 36mg (0.40mmol) of 4-aminobutanol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 60mg (0.2mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 16 h.

The crude product was purified by HPLC to yield 44mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆)：[ppm]＝8.46(s,1H),8.16(d,1H),8.09(s,1H),7.62(d,1H),7.56(s,1H),7.29(d,1H),7.01(d,1H),6.94(dd,1H),4.53(t,2H),3.84(s,3H),2.80(t,2H),1.97-1.87(m,2H),1.76-1.66(m,2H)。

LC-MS (method 1): r_t＝0.78min；MS(ESIpos)m/z＝353[M+H]⁺。

Example 9

4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -N, N-dimethylbut-1-amine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 56mg (0.467mmol) of 4- (dimethylamino) butan-1-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 72 h.

The crude product was purified by HPLC to yield 26mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.12(s,1H),7.87(d,1H),7.51(d,1H),7.44(d,1H),7.10(d,1H),6.91(dd,1H),6.76(d,1H),4.54(t,2H),3.89(s,3H),2.45-2.37(m,2H),2.28(s,6H),2.03-1.91(m,2H),1.81-1.69(m,2H)。

LC-MS (method 1): r_t＝0.81min；MS(ESIpos)m/z＝381[M+H]⁺。

Example 10

N, N-diethyl-4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } pentan-1-amine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 76mg (0.467mmol) of 5- (diethylamino) pentan-2-ol were slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 18 h and at 50 ℃ for 24 h.

The crude product was purified by HPLC to give 23mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.11(s,1H),7.86(d,1H),7.50(d,1H),7.39(s,1H),7.10(d,1H),6.91(dd,1H),6.72(d,1H),5.39-5.24(m,1H),3.89(s,3H),2.60(quin,6H),1.97-1.60(m,4H),1.53(d,3H),1.04(t,6H)。

LC-MS (method 1): r_t＝1.30min；MS(ESIpos)m/z＝300[M+H]⁺。

Example 11

3- (6-methoxy-1-benzofuran-2-yl) -6- [2- (1-methylpyrrolidin-2-yl) ethoxy ] imidazo [1,2-b ] pyridazine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 62mg (0.467mmol) of 2- (1-methylpyrrolidin-2-yl) ethanol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 18 h.

The crude product was purified by HPLC to yield 46mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.12(s,1H),7.88(d,1H),7.50(d,1H),7.45(d,1H),7.10(d,1H),6.91(dd,1H),6.76(d,1H),4.67-4.50(m,2H),3.89(s,3H),3.19-3.09(m,1H),2.41(s,3H),2.39-2.29(m,2H),2.29-2.18(m,1H),2.16-2.02(m,1H),1.95-1.58(m,4H)。

LC-MS (method 1): r_t＝0.77min；MS(ESIpos)m/z＝393[M+H]⁺。

Example 12

1- (2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } ethyl) imidazolidin-2-one

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 62mg (0.467mmol) of 1- (2-hydroxyethyl) imidazolidin-2-one are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 17 h.

The reaction mixture was carefully poured into water. The precipitate was filtered and washed with water to give 81mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆)：[ppm]＝8.12(d,1H),8.05(s,1H),7.63-7.55(m,2H),7.24(s,1H),6.98(d,1H),6.89(dd,1H),6.38(s,1H),4.56(t,2H),3.87-3.74(m,3H),3.56(t,2H),3.51-3.42(m,2H),3.25-3.16(m,2H)。

LC-MS (method 1): r_t＝0.97min；MS(ESIpos)m/z＝394[M+H]⁺。

Example 13

3- (6-methoxy-1-benzofuran-2-yl) -6- [3- (morpholin-4-yl) propoxy ] imidazo [1,2-b ] pyridazine

16mg (0.41mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2mL of anhydrous THF in an ice bath. 71mg (0.467mmol)3- (morpholin-4-yl) propan-1-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 70mg (0.23mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at room temperature for 18 h.

The crude product was purified by HPLC to give 19mg of the title compound.

¹H-NMR(300MHz,CDCl₃)：[ppm]＝8.12(s,1H),7.88(d,1H),7.50(d,1H),7.44(s,1H),7.10(d,1H),6.91(dd,1H),6.76(d,1H),4.58(t,2H),3.89(s,3H),3.79-3.70(m,4H),2.61(t,2H),2.57-2.49(m,4H),2.19-2.06(m,2H)。

LC-MS (method 1): r_t＝0.77min；MS(ESIpos)m/z＝409[M+H]⁺。

Example 14

N- [ (2R) -2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide

To 50mg (0.136mmol) of N- [ (2R) -2- { [3- (6-hydroxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -propyl ] acetamide (example 5) in 2mL of THF are added 37mg (0.273mmol) of potassium carbonate and 9. mu.L (0.15mmol) of iodomethane. The reaction mixture was stirred at room temperature for 12 hours. 9 μ L (0.15mmol) of iodomethane were again added and stirring was continued at room temperature for 5 hours. A third 9. mu.L (0.15mmol) of iodomethane was added and stirring was continued at room temperature for 1 hour. Brine was added. The mixture was extracted with ethyl acetate. The combined organic layers were concentrated and the resulting crude product was purified by HPLC to give 24mg of the title compound.

¹H NMR(400MHz,DMSO-d₆)[ppm]＝8.09-8.18(m,3H),7.63(d,1H),7.58(d,1H),7.29(d,1H),6.92-6.98(m,2H),5.31-5.42(m,1H),3.85(s,3H),3.42-3.55(m,2H),1.82(s,3H),1.44(d,3H)。

LC-MS (method 2): r_t＝0.1.02min；MS(ESIpos)m/z＝381[M+H]⁺。

Example 15

(2R) -2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine

14.5mg (0.36mmol) of sodium hydride (60% dispersion in mineral oil) are dispersed in 2.4mL of anhydrous THF in an ice bath. 27mg (0.47mmol) of (2R) -1-aminopropan-2-ol are slowly added. After the addition was complete, stirring was continued at 0 ℃ for 15 minutes. 78mg (0.26mmol) 6-chloro-3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazine (intermediate 2) are added, the ice bath is removed and the resulting mixture is stirred at 60 ℃ for 16 h.

The crude product was purified by HPLC to give 19mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆)：[ppm]＝8.30(s,1H),8.18(d,1H),8.11(s,1H),7.63(d,1H),7.52(s,1H),7.30(d,1H),6.99(d,1H),6.95(dd,1H),5.38-5.28(m,1H),3.85(s,3H),3.04(d,2H),1.49(d,3H)。

LC-MS (method 1): r_t＝0.79min；MS(ESIpos)m/z＝339[M+H]⁺。

Furthermore, the compounds of general formula (I) of the present invention may be converted into any of the salts described herein by any method known to those skilled in the art. Likewise, any salt of a compound of formula (I) of the present invention may be converted to the free compound by any method known to those skilled in the art.

Pharmaceutical compositions of the compounds of the invention

The invention also relates to pharmaceutical compositions comprising one or more compounds of the invention. These compositions can be used to achieve a desired pharmacological effect by administration to a patient in need thereof. For the purposes of the present invention, a patient is a mammal, including a human, in need of treatment for a particular condition or disease. Accordingly, the present invention includes pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound of the present invention or a salt thereof. A pharmaceutically acceptable carrier is preferably one that is relatively non-toxic and non-injurious to a patient at concentrations consistent with effective activity of the active ingredient, such that any side effects caused by the carrier do not destroy the beneficial effects of the active ingredient. A pharmaceutically effective amount of a compound is preferably an amount that results in or affects the particular condition being treated. The compounds of the present invention may be administered together with pharmaceutically acceptable carriers known in the art in any effective conventional dosage unit form, including immediate release, sustained release and timed release formulations, in the following manner: oral, parenteral, topical, nasal, ocular (opthalmologicaly), ocular (opthalcogly), sublingual, rectal, vaginal, and the like.

For oral administration, the compounds may be formulated into solid or liquid preparations such as capsules, pills, tablets, troches (troche), lozenges (lozenge), melt gels (melt), powders, solutions, suspensions or emulsions and may be prepared according to methods known in the art for the preparation of pharmaceutical compositions. The solid unit dosage form may be a capsule, which may be of the ordinary hard or soft capsule type, comprising, for example, surfactants, lubricants, and inert fillers such as lactose, sucrose, calcium phosphate, and corn starch.

In another embodiment, the compounds of the present invention can be compressed into tablets in combination with conventional tablet bases (e.g., lactose, sucrose and corn starch) and the following: binders such as acacia, corn starch or gelatin, disintegrating agents to aid disintegration and dissolution of the tablet after administration, such as potato starch, alginic acid, corn starch and guar gum, tragacanth, acacia, lubricants to improve the flowability of the tablet particles and to prevent adhesion of the tablet material to the surfaces of the tablet die and punch, such as talc, stearic acid or magnesium stearate, calcium or zinc stearate, dyes, colorants and flavouring agents to improve the organoleptic properties of the tablet and make them more acceptable to the patient, such as peppermint, oil of wintergreen or cherry flavouring. Suitable excipients for oral liquid dosage forms include dicalcium phosphate and diluents such as water and alcohols (e.g., ethanol, benzyl alcohol, and polyethylene glycol), with or without the addition of pharmaceutically acceptable surfactants, suspending agents or emulsifying agents. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets, pills, or capsules may be coated with shellac, sugar or both.

Dispersible powders and granules are suitable for use in the preparation of an aqueous suspension. They provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Examples of suitable dispersing or wetting agents and suspending agents are those mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, such as those described above, may also be present.

The pharmaceutical composition of the invention may also be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil, such as liquid paraffin, or a mixture of vegetable oils. Suitable emulsifying agents may be (1) natural gums, for example gum acacia and gum tragacanth, (2) natural phosphatides, for example soya bean lecithin and lecithin, (3) esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, (4) condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsion may also contain sweetening and flavoring agents.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. The suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate; one or more colorants; one or more flavoring agents; and one or more sweetening agents, such as sucrose or saccharin.

Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent and a preservative such as methyl and propyl parabens as well as flavoring and coloring agents.

The compounds of the invention may also be administered parenterally, i.e., subcutaneously, intravenously, intraocularly, intrasynovially, intramuscularly or intraperitoneally, in an injectable form of the compound, preferably in a physiologically acceptable diluent with a pharmaceutical carrier, which may be a sterile liquid or a mixture of liquids, such as water, saline, aqueous dextrose and related sugar solutions, alcohols such as ethanol, isopropanol or cetyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2, 2-dimethyl-1, 1-dioxolane-4-methanol, ethers such as poly (ethylene glycol) 400, oils, fatty acids, fatty acid esters or glycerides or acetylated glycerides, with or without the addition of pharmaceutically acceptable surfactants such as soaps or detergents, suspending agents such as pectin, carbomer, methylcellulose, hypromellose or carboxymethylcellulose, or emulsifying agents and other pharmaceutically acceptable adjuvants.

Exemplary oils useful in the parenteral formulations of the invention are those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum and mineral oil. Suitable fatty acids include oleic acid, stearic acid, isostearic acid and myristic acid. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate. Suitable soaps include fatty acid alkali metal, ammonium and triethanolamine salts, and suitable detergents include cationic detergents such as dimethyl dialkyl ammonium halides, alkyl pyridinium halides and alkylamine acetates; anionic detergents such as alkyl sulfonates, aryl sulfonates and olefin sulfonates, alkyl sulfates and alkyl sulfosuccinates, olefin sulfates and olefin sulfosuccinates, ether sulfates and ether sulfosuccinates, and monoglyceride sulfates and monoglycerides sulfosuccinates; nonionic detergents such as fatty amine oxides, fatty acid alkanolamides, and poly (oxyethylene-oxypropylene) or ethylene oxide copolymers or propylene oxide copolymers; and amphoteric detergents such as alkyl-beta-aminopropionates and 2-alkylimidazoline quaternary ammonium salts, and mixtures thereof.

The parenteral compositions of the invention will typically comprise from about 0.5% to about 25% by weight of the active ingredient in solution. Preservatives and buffers may also be advantageously employed. To minimize or eliminate irritation at the injection site, such compositions may comprise a nonionic surfactant having a hydrophilic-lipophilic balance (HLB) of preferably from about 12 to about 17. The amount of surfactant in such formulations is preferably from about 5% to about 15% by weight. The surfactant may be a single component having the above HLB, or a mixture of two or more components having the desired HLB.

Exemplary surfactants for parenteral formulations are polyethylene sorbitan fatty acid esters such as sorbitan monooleate, and the high molecular weight adducts of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide and propylene glycol.

The pharmaceutical composition may be in the form of a sterile aqueous suspension for injection. Such suspensions may be formulated according to known methods using: suitable dispersing or wetting agents and suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hypromellose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents which may be a naturally-occurring phosphatide, for example lecithin, condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyoxyethylene sorbitan monooleate.

The sterile injectable preparation may also be a sterile solution or suspension for injection in a non-toxic parenterally-acceptable diluent or solvent. Diluents and solvents which can be used are, for example, water, ringer's solution, isotonic sodium chloride solution and isotonic glucose solution. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. In this regard, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compositions of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are, for example, cocoa butter and polyethylene glycols.

Another formulation used in the methods of the invention utilizes a transdermal delivery device ("patch"). Such transdermal patches may be used to provide continuous or discontinuous delivery of a controlled amount of a compound of the present invention. The construction and use of transdermal patches for delivering agents is well known in the art (see, e.g., U.S. patent No.5,023,252 issued on 6/11 of 1991, which is incorporated herein by reference). Such patches may be configured for continuous, pulsed, or on-demand delivery of the agent.

Controlled release formulations for parenteral administration include liposomal microspheres, polymeric microspheres, and polymeric gel formulations known in the art.

It may be desirable or necessary to deliver the pharmaceutical composition to a patient by a mechanical delivery device. The construction and use of mechanical delivery devices for delivering pharmaceutical agents is well known in the art. Direct techniques such as administering drugs directly to the brain typically involve placing a drug delivery catheter into the ventricular system of the patient to bypass the blood brain barrier. One such implantable delivery system for delivering agents to specific anatomical locations of the body is described in U.S. patent No.5,011,472 issued on 30/4 1991.

The compositions of the present invention may also contain, as necessary or desired, other conventional pharmaceutically acceptable formulation ingredients, which are commonly referred to as carriers or diluents. Conventional procedures for preparing such compositions into suitable dosage forms may be used. Such ingredients and procedures include those described in the following references, all of which are incorporated herein by reference: powell, M.F. et al, "Complex of Excipients for particulate Formulations" PDA Journal of pharmaceutical Science & Technology 1998,52(5), 238-; strickley, R.G, "partial formulations of Small Molecule Therapeutics marked in the United States (1999) -Part-1," PDA Journal of Pharmaceutical Science & Technology 1999,53(6), 324-; and Nema, S. et al, "Excipients and the same Use in Injectable Products," PDAjournal of Pharmaceutical Science & Technology 1997,51(4),166- "171.

Common pharmaceutical ingredients that may be used to formulate the composition for the intended route of administration include:

acidulants (examples include, but are not limited to, acetic acid, citric acid, fumaric acid, hydrochloric acid, nitric acid);

alkalizing agents (examples include, but are not limited to, aqueous ammonia, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium hydroxide, triethanolamine (triethanolamine), triethanolamine (trolamine));

adsorbents (examples include, but are not limited to, powdered cellulose and activated carbon);

aerosol propellants (examples include, but are not limited to, carbon dioxide, CCl₂F₂、F₂ClC-CClF₂And CClF₃)；

Air displacement agents (examples include, but are not limited to, nitrogen and argon);

antifungal preservatives (examples include, but are not limited to, benzoic acid, butyl paraben, ethyl paraben, methyl paraben, propyl paraben, sodium benzoate);

antibacterial preservatives (examples include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, phenylmercuric nitrate, and thimerosal);

antioxidants (examples include, but are not limited to, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorous acid, thioglycerol, propyl gallate, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite);

adhesive substances (examples include, but are not limited to, block polymers, natural and synthetic rubbers, polyacrylates, polyurethanes, silicones, polysiloxanes, and styrene-butadiene copolymers);

buffering agents (examples include, but are not limited to, potassium metaphosphate, dipotassium hydrogen phosphate, sodium acetate, anhydrous sodium citrate, and sodium citrate dihydrate);

a carrier (examples include, but are not limited to, acacia syrup, flavoring elixir, cherry syrup, cocoa syrup, orange syrup, corn oil, mineral oil, peanut oil, sesame oil, bacteriostatic sodium chloride injection, and bacteriostatic water for injection);

chelating agents (examples include, but are not limited to, sodium edetate and edetic acid);

coloring agents (examples include, but are not limited to FD & C Red No.3, FD & C Red No.20, FD & C Yellow No.6, FD & C Blue No.2, D & C Green No.5, D & C Orange No.5, D & C Red No.8, caramel, and Red iron oxide);

clarifying agents (examples include, but are not limited to, bentonite);

emulsifying agents (examples include, but are not limited to, acacia, cetomacrogol, cetyl alcohol, glyceryl monostearate, lecithin, sorbitan monooleate, polyoxyethylene 50 monostearate);

encapsulating agents (examples include, but are not limited to, gelatin and cellulose acetate phthalate);

flavors (examples include, but are not limited to, anise oil, cinnamon oil, cocoa, menthol, orange oil, peppermint oil, and vanillin);

humectants (examples include, but are not limited to, glycerin, propylene glycol, and sorbitol);

abrasives (examples include, but are not limited to, mineral oil and glycerin);

oils (examples include, but are not limited to, peanut oil (arachis oil), mineral oil, olive oil, peanut oil (parautoil), sesame oil, and vegetable oils);

ointment bases (examples include, but are not limited to, lanolin, hydrophilic ointment, polyethylene glycol ointment, petrolatum, hydrophilic petrolatum, white ointment, yellow ointment, and rose water ointment);

penetration enhancers (transdermal delivery) (examples include, but are not limited to, mono-or polyhydric alcohols, mono-or polyvalent alcohols, saturated or unsaturated fatty esters, saturated or unsaturated dicarboxylic acids, essential oils, phosphatidyl derivatives, cephalins, terpenes, amides, ethers, ketones, and ureas);

plasticizers (examples include, but are not limited to, diethyl phthalate and glycerol);

solvents (examples include, but are not limited to, ethanol, corn oil, cottonseed oil, glycerol, isopropanol, mineral oil, oleic acid, peanut oil, purified water, water for injection, sterile water for injection, and sterile water for rinsing);

hardening agents (examples include, but are not limited to, cetyl alcohol, cetyl esters wax, microcrystalline wax, paraffin, stearyl alcohol, white wax, and yellow wax);

suppository bases (examples include, but are not limited to, cocoa butter and polyethylene glycol (mixtures));

surfactants (examples include, but are not limited to, benzalkonium chloride, nonoxynol 10, octoxynol 9, polysorbate 80, sodium lauryl sulfate, and sorbitan monopalmitate);

suspending agents (examples include, but are not limited to, agar, bentonite, carbomer, sodium carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hypromellose, kaolin, methylcellulose, tragacanth and magnesium aluminum silicate);

sweetening agents (examples include, but are not limited to, aspartame, dextrose, glycerin, mannitol, propylene glycol, saccharin sodium, sorbitol, and sucrose);

tablet antiadherents (examples include, but are not limited to, magnesium stearate and talc);

tablet binders (examples include, but are not limited to, acacia, alginic acid, sodium carboxymethylcellulose, compressible sugar, ethylcellulose, gelatin, liquid glucose, methylcellulose, non-crosslinked polyvinylpyrrolidone, and pregelatinized starch);

tablet and capsule diluents (examples include, but are not limited to, dibasic calcium phosphate, kaolin, lactose, mannitol, microcrystalline cellulose, powdered cellulose, precipitated calcium carbonate, sodium phosphate, sorbitol, and starch);

tablet coatings (examples include, but are not limited to, liquid glucose, hydroxyethyl cellulose, hydroxypropyl cellulose, hypromellose, methyl cellulose, ethyl cellulose, cellulose acetate phthalate, and shellac);

tablet direct compression excipients (examples include, but are not limited to, dibasic calcium phosphate);

tablet disintegrating agents (examples include, but are not limited to, alginic acid, carboxymethylcellulose calcium, microcrystalline cellulose, polacrillin potassium, crospovidone, sodium alginate, sodium starch glycolate, and starch);

tablet glidants (examples include, but are not limited to, colloidal silicon dioxide, corn starch, and talc);

tablet lubricants (examples include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, stearic acid, and zinc stearate);

tablet/capsule opacifiers (examples include but are not limited to titanium dioxide);

tablet polishes (examples include, but are not limited to, carnauba wax and white wax);

thickening agents (examples include, but are not limited to, beeswax, cetyl alcohol, and paraffin wax);

tonicity agents (examples include, but are not limited to, glucose and sodium chloride);

viscosity increasing agents (examples include, but are not limited to, alginic acid, bentonite, carbomer, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, sodium alginate, and gum tragacanth); and

wetting agents (examples include, but are not limited to, heptadecaethyleneoxycetanol, lecithin, sorbitol monooleate, polyoxyethylene sorbitol monooleate, and polyoxyethylene stearate).

The pharmaceutical composition of the present invention can be exemplified as follows:

sterile intravenous solution: sterile water for injection can be used to prepare a 5mg/mL solution of the desired compound of the invention, with the pH adjusted as necessary. The solution was diluted to 1-2mg/mL with sterile 5% glucose for administration and administered as an intravenous infusion over about 60 minutes.

Lyophilized powder for intravenous administration: sterile preparations can be prepared from (i)100-1000mg of the desired compound of the invention in the form of a lyophilized powder, (ii)32-327mg/mL sodium citrate, and (iii)300-3000mg dextran 40. The formulation is reconstituted to a concentration of 10-20mg/mL with sterile saline for injection or 5% glucose, then further diluted to 0.2-0.4mg/mL with saline or 5% glucose and administered as an intravenous bolus or intravenous infusion over 15-60 minutes.

Intramuscular injection suspension: the following solutions or suspensions can be prepared for intramuscular injection:

50mg/mL of the desired Water-insoluble Compound of the invention

5mg/mL sodium carboxymethylcellulose

4mg/mL TWEEN 80

9mg/mL sodium chloride

9mg/mL benzyl alcohol

Hard capsules: by filling each with 100mg of powdered active ingredient, 150mg of lactose, 50mg of cellulose and 6mg of magnesium stearateA standard two-piece hard capsule is filled to make a large number of unit capsules.

Soft capsule: a mixture of the active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by a positive displacement pump into molten gelatin to form soft capsules containing 100mg of the active ingredient. The capsules were washed and dried. The active ingredient may be dissolved in a mixture of polyethylene glycol, glycerol and sorbitol to prepare a water-miscible drug mixture.

Tablet formulation: a number of tablets were prepared by conventional procedures such that the dosage unit contained 100mg of active ingredient, 0.2mg of colloidal silicon dioxide, 5mg of magnesium stearate, 275mg of microcrystalline cellulose, 11mg of starch and 98.8mg of lactose. Suitable aqueous and non-aqueous coatings may be employed to increase palatability, improve appearance and stability, or delay absorption.

Immediate release tablet/capsule: these are solid oral dosage forms prepared by conventional and novel processes. These unit dosage forms are administered orally without the need for water for immediate dissolution and delivery of the drug. The active ingredient is mixed in a liquid containing ingredients such as sugar, gelatin, pectin and sweeteners. These liquids are solidified into solid tablets or caplets by freeze-drying and solid-state extraction techniques. The pharmaceutical compound can be tableted with a viscoelastic and thermoelastic sugar and a polymer or effervescent component to produce a porous matrix that is quick-releasing without the need for water.

Combination therapy

The compounds of the present invention may be administered as the sole agent or in combination with one or more other agents, wherein the combination does not cause unacceptable adverse effects. The invention also relates to such combinations. For example, the compounds of the present invention may be combined with known agents and the like that are resistant to hyperproliferative diseases or other indications, as well as mixtures and combinations thereof. Other indications include, but are not limited to, anti-angiogenic agents, mitotic inhibitors, alkylating agents, anti-metabolites, DNA-intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors, biological response modifiers, or anti-hormones.

According to one embodiment, the present invention relates to a pharmaceutical combination comprising:

-one or more first active ingredients selected from compounds of general formula (I) as defined above, and

-one or more second active ingredients selected from chemotherapeutic anti-cancer agents.

The term "chemotherapeutic anti-cancer agents" includes, but is not limited to:

131I-chTNT, abarelix, abiraterone, aclarubicin, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alemtuzumab, alendronic acid, alitretinol, altretamine, amifostine, aminoglutethimide, Hexyl methylaminoketone valerate (Hexyl aminolevulinate), amrubicin, amsacrine, anastrozole, acesulfame, anethole disulfide (anethole disulfoethionne), angiotensin II, antithrombin III, aprepitant, acipimox, Arglabin, diarsenic oxide, asparaginase, acitinib, azacitidine, basiliximab, belveludoxin, belvelurtan, bicalutamide, bisanthrene (), lebertinib, bortezomib, sultrinib, tribenuron acetate, calcium picatinib (tetrahydrocarb), calcium ascorbate, blevacarbacin, sulbactam acetate, betulin, calcium ascorbate, and bromhexetil acetate, Levofolinic acid calcium, capecitabine, carpolizumab, carboplatin, capeline, capecitabine, carmofur, carmustine, rituximab, celecoxib, simethionine, ceritinib, cetuximab, chlorambucil, progesterone chloride, mechlorethamine, cidofovir, cinacalcet, cisplatin, cladribine, clorphosphoid, clofarabine, Copanlisib, kristine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, actinomycin, dabecopotin alpha, dabrafenib, dasatinib, orthopyristin, decitabine, degarelix, dinil-toxin linker (denileukinindifidititox), didanoxemae (denosumab), deprenopeptide, sertraline, dexrazine, dibromospirobichloroammonium, dianhydrogalactitol, diclofenac (diclofenac), doxorafosalxolone, docetaxel, doxorabicolone + doxorabicubicin, doxorabicubicin + doxorabicubicin, doxorabicubicin + doxorabicubin, Dronabinol, eculizumab (eculizumab), eculizumab (edrecolomab), hydroxypyrazole acetate, eltrombopag, endostatin, enocitabine, enzalutamide, epirubicin, epithiandrol, epoetin α, epoetin β, erythropoietin (epoetin zeta), eptaplatin, eribulin (eribulin), erlotinib, esomeprazole, estradiol, estramustine, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, flumetsterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestramustine, gadobutrol, gadolithic, gadoteridol, gadoteric acid meglumine (gadoteracid), gadovaverine, gadoteridol, gadoteracid, gadoteridol, gracilalone, granulocyte colony stimulating factor, histamine dihydrochloride, histrelin, hydroxyurea, I-125 particles, lansoprazole, ibandronic acid, temozolomide, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronate, ingenol mebutate, interferon alpha, interferon beta, interferon gamma, iobitol, iobenguanide (123I), iomeprol, ipilimumab, irinotecan, itraconazole, ixabepilone, lanreotide, lapatinib, Lasochloline, Railidomide, Reynograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, meglumine, megestrol, gehead, ge, Melphalan, metandienone, mercaptopurine, mesna, methadone, methotrexate, methoxsalene, aminolevulinic acid methyl ester (methyalaminolevulinate), methylprednisolone, methyltestosterone, metirane, mivampicin, miltefosine, miriplatin, dibromomannitol, mitoguazone, dibromodulcitol, mitomycin, mitotane, mitoxantrone, Mogamulizumab, moraxentin, mopidamol, morphine hydrochloride, morphine sulfate, marijuone, Nabiximols, nafarelin, naloxone + tebucsin, naltrexone, nitudin, nedaplatin, nelarabine, neridronic acid, Nivolumapentetretone, nilutamide, nimorazole, nimotuzumab, pyrimidinium, guanidium, nitazomab, tamarine, oxtamide, octreotide, tritaethionine, oximulin, oxaliplatin, orimulin, orimulukin, gulipin, and platinum, Oxycodone, oxymetholone, Ozogamicine, p53 gene therapy, paclitaxel, palifermin, palladium-103 particles, palonosetron, pamidronic acid, panitumumab, pantoprazole, palozolomide, asparaginase, PEG-epoetin beta (methoxy PEG-epoetin beta), pamirumab, pefilgrastim, PEG interferon alpha-2 b, pemetrexed, pentazocine, pentostatin, pellomycin, perfluorobutane, superphosphoramide, pertuzumab, streptolysin, pilocarpine, pirarubicin, Pixantrone, pleroxafungin, mithramycin, chitosan, polyesoryl phosphate, polyvinylpyrrolidone + sodium hyaluronate, coriolus versicolor-K, pamamelidomide, panitudinium, porphyrinum sodium, pralatrexate, prednimone, procarbazine, procarbazole, proparaxolone, quinaldine, p53 gene therapy, paclitaxel, prasuzurine, pemphigomisin, pemetrexendine, pemphigomisin, pemetrexe, Rabeprazole, Racotumomab, radium-223 chloride, ladostinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ramosestine, labyrine, ranibazam, Refametinib, regorafenib, risedronic acid, rhenium Re186 etidronate, rituximab, romidepsin, romopeptide, Ronicolilb, Lyricontinane (153Sm), Samomentin, Satuzumab, secretin, Sipuleucel-T, Sipureocel, Sizopyran, Sobucazoxan, Glycobinazole sodium, sorafenib, Parizolone, streptozocin, sunitinib, talaporfin, Tamibaroreceptor, tamoxifen, Tavoxamine, Tagenin, Texidulin, Mercaptitumomab (99mTc) mercaptol, 99 mTc-3-titutesin, Timentine + Tegazetimothricin, Tegazine + Oxalidomide, Tegiline + Tegiline, Tegiline + Tegilicarb + Tegiline, Teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alpha, thioguanine, tositumumab, topotecan, toremifene, tositumomab, trabectedin, tramadol, trastuzumab-Emtansine conjugate (trastuzumab Emtansine), troosulfan, tretinoid, trifluridine + thymidine phosphorylase inhibitor (trifluridine + tipiracil), trolsartan, triptorelin, trametin, trometinib, trofosfamide, thrombopoietin, tryptophan, ubenix, vartananib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinneat, vinorelbine, vorinostat, vorozole, yttrium-90 glass microspheres, setate, sethoxystatin, zolmithramycin, zolamide, or combinations thereof.

The additional agent may be everolimus, aldesleukin, alendronic acid, alpha-interferon, alitretinoin, allopurinol, sodium allopurinol for injection (alprimem), palonosetron hydrochloride injection (aloxi), altretamine, aminoglutethimide, amifostine, amsacrine, anastrozole, dolasemet, alfa-bepotastine injection (aranesp), parthenolide derivative (arglabin), diarsenic trioxide, arnoxine, 5-azacitidine, azathioprine, BAY 80-6946, BCG or tide, BCG, amastatin, betamethasone acetate, betamethasone sodium phosphate, bexarotene, bleomycin sulfate, uridine bromide, bortezomib, busulfan, calcitonin, argatron (caepath), capecitabine, platinum, pemetrexene, bicolone, bicuccine, daunorubicin, carmustine, doxycycline, and other, Cisplatin, cladribine, clodronic acid, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin liposome citrate (DaunoXome), dexamethasone sodium phosphate, estradiol valerate, dinil interleukin 2, methylprednisolone, deslorelin, dexrazoxane, diethylstilbestrol, tolbutan, docetaxel, doxifluridine, doxorubicin, dronabinol, DW-166HC, leuprolide acetate (eligard), labyrinase injection (elitek), epirubicin hydrochloride injection (ellence), aprepitant capsule (emide), epirubicin, alfa eptin alfa, alfa farabin (epogen), epta, levamisole, estradiol formulations (estratriol), estradiol, sodium phosphate estramustine, amifostine, etidronic acid, piridopipons (etophos), etoposide, posaconazole, fafavudine, favudine, dox, etoposide, doxepidopton, doxorfazole, doxepirubin, doxycycline, doxyc, Filgrastim, finasteride, filgrastim, floxuridine, tolbutamide, fludarabine, 5-fluorodeoxyuridine monophosphate, 5-fluorouracil (5-FU), fluoxymesterone, flutamide, formestane, fosetabine, fotemustine, fulvestrant, gamma-globulin (gammagard), gemcitabine, gemumab, gleevec, carmine, carmustine capsules (gliadel), goserelin, granisetron hydrochloride, histrelin, and metin, hydrocortisone, erythrohydroxynonyladenine (eyrtho-hydroxyynyladenine), hydroxyurea, temomamab, idarubicin, ifosfamide, alpha interferon, alpha 2 interferon, alpha-2A interferon, alpha-2B interferon, alpha-n 1 interferon, alpha-n 3 interferon, beta interferon, gamma-1 a interferon, interleukin 2A, alpha-intron A (alpha-Intron A), Iressa, irinotecan, Ketirel, lapatinib, lentinan sulfate, letrozole, leucovorin, leuprorelin acetate, levamisole, calcium levofolinate, levothyroxine sodium (levothroid), levothyroxine sodium preparation (levoxyl), lomustine, lonidamine, dronabinol, mechlorethamine, mecobalamin acetate, megestrol acetate, melphalan, esterified estrogen preparation (menest), 6-mercaptopurine, mesna, methotrexate, medecine, miltefosine, minocycline, mitomycin C, mitotane, mitoxantrone, trostan (Modrenal), doxorubicin liposomes (Myocet), nedaplatin, filgrastimostatin (Entadat), recombinant human interleukin 11 (neuglada), Yogutan (Neugen), Neuguega, Neugueger, Neuguent, Neiwan, Neiwatt, Novavas, OCT-8943, Tritriptan-631570, OCT-C-631570, Tritretin (Octrexate, and its) and its preparation, Ondansetron hydrochloride, prednisolone oral rapidly disintegrating tablet (orapred), oxaliplatin, paclitaxel, prednisone sodium phosphate preparation (pediapared), pemetrexed, pyroxene, pentostatin, streptolysin preparation, pilocarpine hydrochloride, pirarubicin, plicamycin, porfimer sodium, prednisolone, prednisone, bemeili, procarbazine, recombinant human erythropoietin alpha, raltitrexed, RDEA 119, recombinant human interferon beta 1a injection (rebif), rhenium-186 hydroxyethylphosphonate, rituximab, roscovitine (roferon-A), romopeptide, pilocarpine hydrochloride tablet (salagen), shannin, sargrastim, semustine, Sizopyran, sobuzosin, methylprednisolone, fosetyl-aspartic acid, dry cell therapy, zosin, strontium 89, sunitinib, levofloxacin, thyroxine, neoxiphenytoin, and neotamicin, Tasolomine, testolactone, docetaxel injection (taxotere), teicoplanin, temozolomide, teniposide, testosterone propionate, mesterolone capsule (tetred), thioguanine, thiotepa, thyrotropin, tiludronic acid, topotecan, toremifene, tositumomab, trastuzumab, troosulfan, tretinoin, methotrexate (trexal), trimethylmelamine, trimetrexate, triptorelin acetate, triptorelin pamoate, UFT, uridine, valrubicin, vesnarinone, vinblastine, vincristine, vindesine, vinorelbine, vilulizine, dexrazol, dexrazoxane, setastine, pinocetin, abine, ABI-007, allophile (aclobirene), interferon gamma-1 b (actimmumole), finafiltrene, aminopterin, arzoxifene, progesterone receptor modulator (ioprisperidone), oxypetamol (9006-9006) and bafenib (oxypheniramine, oxyphenirane, ketonura-43) Avastin (Avastin), CCI-779, CDC-501, celecoxib, cetuximab, clinacator, cyproterone acetate, decitabine, DN-101, doxorubicin-MTC, dSLIM, dutasteride, edotecarin, efluoromidine, efaprotecan, fenretinide, histamine dihydrochloride, histrelin hydrogel implants, holmium-166 DOTMP, ibandronic acid, gamma interferon, pegylated interferon alpha-2 b (intron-PEG), ixabepilone, keyhole limpet hemocyanin (keyhol limpet hemocyanin), L-651582, lanreotide, lasofoxifene, libra, farnesol protein transferase inhibitors (lonarnib), mirepredfene, minophosphonate (minodronate), MS-209, liposome-PE, MX-6, nararelin, laniroxostat, nemorosa, neosloxostat, minostrobin, trimethoprim, celecoxib, medroxypterin, medroxypr S, merrimeric S, merremicade, neritin, nereid, doxylamine, and doxylamine, Paclitaxel, disodium pamidronate, PN-401, QS-21, quasipam, R-1549, raloxifene, ranpirnase, 13-cis-retinoic acid, satraplatin, seocalcitol, T-138067, Tarceva (tarceva), taxoprexin, alpha-1 thymosin, thiazolufrine, tipifarnib, tirapazamine, TLK-286, toremifene, TransMID-107R, valcephrade, vapreotide, vatalanib (vatalanib), verteporfin, vinflunine, Z-100, zoledronic acid, or combinations thereof.

Optional anti-hyperproliferative agents that may be added to the compositions include, but are not limited to, compounds listed in the cancer chemotherapeutic regimen of the Merck index 11 edition (1996) (incorporated herein by reference), such as asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, levoasparaginase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin (adriamycin), epirubicin, epothilone derivatives, etoposide, 5-fluorouracil, hexamethylmelamine, hydroxyurea, ifosfamide, irinotecan, leucovorin, lomustine, nitrogen mustard, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone, prednisone, procarbazine, raloxifene, streptozocin, tamoxifen, meclizine, thioguanine, topotecan, vinblastine, vincristine, and vindesine.

Other anti-hyperproliferative agents suitable for use with The compositions of The present invention include, but are not limited to, those compounds recognized for use in The treatment of neoplastic diseases in Goodmanand Gilman's, The Pharmacological Basis of Therapeutics (9 th edition), edited by Molinoff et al, McGraw-Hill, pp.1225-1287 (1996) (incorporated herein by reference), e.g., aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine, cladribine, busulfan, diethylstilbestrol, 2' -difluorodeoxycytidine, docetaxel, erythrononyl adenine, ethinyl estradiol, 5-fluorodeoxyuridine monophosphate, fludarabine phosphate, fluorometholone, flutamide, hydroxyprogesterone caproate, idarubicin, interferon, medroxyprogesterone acetate, megestrol acetate, melphalan, mitotanetan, medroxyprogesterone, medroxyprogen, medullone, medullol acetate, melphalan, medetotan, and The like, Paclitaxel, pentostatin, N-phosphonoacetyl-L-aspartate (PALA), plicamycin, semustine, teniposide, testosterone propionate, thiotepa, trimethylmelamine, uridine, and vinorelbine.

Other anti-hyperproliferative agents suitable for use with the compositions of the present invention include, but are not limited to, other anti-cancer agents such as epothilone and its derivatives, irinotecan, raloxifene and topotecan.

The compounds of the invention may also be administered in combination with a protein therapeutic. Such protein therapeutics suitable for use in the treatment of cancer or other angiogenic disorders and suitable for use with the compositions of the invention include, but are not limited to, interferons (e.g., alpha, beta, or gamma interferons), hyperactive monoclonal antibodies, Tuebingen, TRP-1 protein vaccines, Colostrinin, anti-FAP antibodies, YH-16, gimumab, infliximab, cetuximab, trastuzumab, dinil interleukin 2, rituximab, alpha 1 thymosin, bevacizumab, mecamylamine Rifafibate (mecamylin Rifabat), Ompur interleukin, natalizumab, rhMBL, MFE-CP1+ ZD-2767-P, ABT-828, ErbB 2-specific immunotoxins, SGN-35, MT-103, Rifamate (rinfabate), AS-1402, B43-genistein, L-19 series radioimmunotherapeutic, AC-9301, NY-ESO-1 vaccine, IMC-1C11, CT-322, rhCC10, r (m) CRP, MORAB-009, Avisuramine (aviscumine), MDX-1307, Her-2 vaccine, APC-8024, NGR-hTNF, rhH1.3, IGN-311, endostatin, Voloximab (volociximab), PRO-1762, lexatuzumab (lexatuzumab), SGN-40, pertuzumab (pertuzumab), EMD-273, L19-IL-2 fusion protein, PRX-321, CNTO-328, MDX-214, tegafur peptide (tigotide), CAT-3888, labetazumab (labetazumab), α -emitting radioisotope cross-linked monoclonal antibody, EM-Acuk-1421, interleukin-14225, interleukin-7, HPV-30625, HPV-3063, and Ab-40, Javelin-melanoma, NY-ESO-1 vaccine, EGF vaccine, CYT-004-MelQbG10, WT1 peptide, agovacizumab (oregomab), ofatumumab (ofatumumab), zalutumumab (zalutumumab), bevacizine (cindrekin bestudox), WX-G250, Albuferon, aflibercept (aflibercept), denozumab (denosumab), vaccine, CTP-37, efungumab (efungumab), or 131I-chTNT-1/B. Monoclonal antibodies useful as protein therapeutics include, but are not limited to, molobuzumab-CD 3, abciximab, edrecolomab, daclizumab, gemtuzumab (gentuzumab), alemtuzumab, ibritumomab tiuxetan (ibritumomab), cetuximab, bevacizumab, efalizumab (efalizumab), adalimumab (adalimumab), omalizumab, moelimumab-CD 3, rituximab, daclizumab, trastuzumab, palivizumab, basiliximab, and infliximab.

The compounds of the invention may also be combined with biological therapeutic agents such as antibodies (e.g., Avastin (Avastin), B cell monoclonal antibodies (Rituxan), erbitux (Erbitax), Herceptin (Herceptin)) or recombinant proteins.

-one or more compounds of general formula (I) above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same; and

-one or more drugs selected from: a taxane, such as docetaxel, paclitaxel, lapatinib, sunitinib, or taxol; epothilones, such as ixabepilone, paclitaxel, or salgopirone; mitoxantrone; prednisolone; dexamethasone; estramustine; vinblastine; vincamine; doxorubicin; doxorubicin; idarubicin; daunorubicin; bleomycin; etoposide; cyclophosphamide; ifosfamide; procarbazine; melphalan; 5-fluorouracil; capecitabine; fludarabine; cytarabine; Ara-C; 2-chloro-2' -deoxyadenosine; thioguanine; antiandrogens, such as flutamide, cyproterone acetate, or bicalutamide; bortezomib; platinum derivatives, such as cisplatin, or carboplatin; chlorambucil; methotrexate; and rituximab.

The compounds of the invention may also be combined with an anti-angiogenic agent, for example with avastin, axitinib, DAST, recentin, sorafenib or sunitinib. It may also be combined with a proteasome inhibitor or an mTOR inhibitor or an anti-hormonal agent or a steroid metabolic enzyme inhibitor.

In general, the use of cytotoxic and/or cytostatic agents in combination with a compound or composition of the invention will serve the following functions:

(1) produces better efficacy in reducing tumor growth or even eliminating tumors than either agent administered alone,

(2) allowing for the administration of smaller amounts of the administered chemotherapeutic agent,

(3) providing a chemotherapeutic treatment that is well tolerated by patients and has fewer harmful pharmacological complications than observed with single agent chemotherapy and certain other combination therapies,

(4) allowing the treatment of a wider range of different cancer types in mammals, particularly humans,

(5) providing a higher response rate in the treated patient,

(6) provides longer survival in the treated patient compared to standard chemotherapy treatment,

(7) provide longer tumor progression time, and/or

(8) At least as good efficacy and tolerability as the agents used alone are obtained as compared to known cases where other cancer agents produce antagonistic effects in combination.

Method for sensitizing cells to radiation

In a different embodiment of the invention, the compounds of the invention can be used to sensitize cells to radiation. That is, treating cells with a compound of the invention prior to radiation therapy of the cells makes the cells more susceptible to DNA damage and cell death than they would be if the cells were not subjected to any treatment with a compound of the invention. In one aspect, a cell is treated with at least one compound of the invention.

Accordingly, the present invention also provides a method of killing cells, wherein one or more compounds of the invention are administered to the cells along with conventional radiation therapy.

The invention also provides a method of rendering a cell more susceptible to cell death, wherein the cell is treated with one or more compounds of the invention to cause or induce cell death prior to treating the cell. In one aspect, after treating the cells with one or more compounds of the invention, the cells are treated with at least one compound or at least one method or a combination thereof to cause DNA damage for inhibiting the function of normal cells or killing the cells.

In one embodiment, the cells are killed by treating the cells with at least one DNA damaging agent. That is, after treating a cell with one or more compounds of the invention to sensitize the cell to cell death, the cell is treated with at least one DNA damaging agent to kill the cell. DNA damaging agents useful in the present invention include, but are not limited to, chemotherapeutic agents (e.g., cisplatin), ionizing radiation (X-ray, ultraviolet radiation), carcinogens, and mutagenic agents.

In another embodiment, the cells are killed by treating the cells with at least one method to cause or induce DNA damage. Such methods include, but are not limited to: activating a cellular signal transduction pathway (which causes DNA damage when the pathway is activated), inhibiting a cellular signal transduction pathway (which causes DNA damage when the pathway is inhibited), and inducing a biochemical change in a cell (wherein the change causes DNA damage). By way of non-limiting example, DNA repair pathways in a cell may be inhibited, thereby preventing repair of DNA damage and resulting in abnormal accumulation of DNA damage in a cell.

In one aspect of the invention, the compounds of the invention are administered prior to irradiation or other induction that causes DNA damage in the cell. In another aspect of the invention, the compounds of the invention are administered concurrently with irradiation or other induction that causes DNA damage to cells. In another aspect of the invention, the compounds of the invention are administered immediately after the initiation of irradiation or other induction that causes DNA damage to the cells.

In another aspect, the cell is in vitro. In another embodiment, the cell is in vivo.

As described above, it has surprisingly been found that the compounds of the present invention effectively inhibit MKNK-1 and are therefore useful for the treatment or prevention of diseases caused by or accompanied by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response, in particular wherein said uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is mediated by MKNK-1, e.g. hematological tumors, solid tumors and/or their metastases, such as leukemia and myelodysplastic syndrome, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, tumors, and/or metastases, in particular, in the treatment or prevention of diseases caused by or accompanied by uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response Breast, gastrointestinal, endocrine, breast and other gynaecological tumours including non-small cell and small cell lung tumours, urological tumours including renal, bladder and prostate tumours, skin tumours and sarcomas, and/or metastases thereof.

Thus, according to another aspect, the present invention relates to a compound of general formula (I), stereoisomers, tautomers, N-oxides, hydrates, solvates, or salts thereof, particularly pharmaceutically acceptable salts, or mixtures thereof, as described and defined herein, for use in the treatment or prevention of a disease as described above.

Thus, another particular aspect of the present invention is the use of a compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, as described above, for the prophylaxis or treatment of a disease.

Accordingly, another particular aspect of the present invention is the use of a compound of general formula (I) as described above for the preparation of a pharmaceutical composition for the treatment or prevention of a disease.

Another aspect of the present invention is the use of a compound of formula (I) as described above for the preparation of a medicament for the prevention or treatment of a disease.

The diseases mentioned in the first three paragraphs are diseases caused by or accompanied by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response, in particular wherein said uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is mediated by MKNK-1, such as hematological tumors, solid tumors and/or metastases thereof, e.g. leukemia and myelodysplastic syndrome, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, breast tumors including non-small cell lung tumors and small cell lung tumors, gastrointestinal tumors, inflammatory diseases, Endocrine tumors, breast tumors and other gynecological tumors, urological tumors including renal tumors, bladder tumors and prostate tumors, skin tumors and sarcomas, and/or metastases thereof.

In the context of the present invention, in particular in the context of an "inappropriate cellular immune response or inappropriate cellular inflammatory response" as used herein, the term "inappropriate" is to be understood as preferably meaning a response which is weaker or stronger than the normal response and which is associated with, causes or leads to the pathology of the disease.

Preferably, the use is for the treatment or prevention of a disease, wherein the disease is a hematological tumor, a solid tumor and/or metastases thereof.

Methods of treating hyperproliferative disorders

The present invention relates to methods of treating hyperproliferative disorders in mammals using the compounds of the present invention and compositions thereof. The compounds may be used to inhibit, block, reduce, etc., cell proliferation and/or cell division and/or induce apoptosis. The method comprises administering to a mammal, including a human, in need thereof an amount of a compound of the present invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof, and the like, effective to treat the condition. Hyperproliferative disorders include, but are not limited to, for example, psoriasis, keloids and other hyperplasia affecting the skin, Benign Prostatic Hyperplasia (BPH), solid tumors such as breast cancer, respiratory tract cancer, brain cancer, reproductive organ cancer, digestive tract cancer, urinary tract cancer, eye cancer, liver cancer, skin cancer, head and neck cancer, thyroid cancer, parathyroid cancer and their distal metastases. Those conditions also include lymphomas, sarcomas, and leukemias.

Examples of breast cancer include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.

Examples of cancers of the respiratory tract include, but are not limited to, small cell lung cancer and non-small cell lung cancer as well as bronchial adenomas and pleural pneumococcal tumors.

Examples of brain cancers include, but are not limited to, brainstem and hypothalamic gliomas, cerebellum and brain astrocytomas, medulloblastomas, ependymomas, and neuroectodermal and pineal tumors.

Tumors of the male reproductive organs include, but are not limited to, prostate cancer and testicular cancer. Tumors of female reproductive organs include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancers, as well as uterine sarcomas.

Tumors of the digestive tract include, but are not limited to, anal, colon, colorectal, esophageal, gallbladder, gastric, pancreatic, rectal, small intestine, and salivary gland cancers.

Urinary tract tumors include, but are not limited to, bladder cancer, penile cancer, kidney cancer, renal pelvis cancer, ureter cancer, urinary tract cancer, and human papillary renal cancer.

Eye cancers include, but are not limited to, intraocular melanoma and retinoblastoma.

Examples of liver cancer include, but are not limited to, hepatocellular carcinoma (with or without fibrolamellar variation), cholangiocarcinoma (intrahepatic cholangiocarcinoma), and mixed hepatocellular cholangiocarcinoma.

Skin cancers include, but are not limited to, squamous cell carcinoma, kaposi's sarcoma, malignant melanoma, merkel cell skin cancer, and non-melanoma skin cancer.

Head and neck cancers include, but are not limited to, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, lip cancer, oral cavity cancer, and squamous cell. Lymphomas include, but are not limited to, aids-related lymphoma, non-hodgkin's lymphoma, cutaneous T-cell lymphoma, burkitt's lymphoma, hodgkin's disease, and central nervous system lymphoma.

Sarcomas include, but are not limited to, soft tissue sarcomas, osteosarcomas, malignant fibrous histiocytomas, lymphosarcomas, and rhabdomyosarcomas.

Leukemias include, but are not limited to, acute myelogenous leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.

These conditions have been well characterized in humans, but also exist in other mammals with similar etiologies, and can be treated by administering the pharmaceutical compositions of the present invention.

The term "treating" as referred to throughout this document generally manages or cares for the patient, e.g., for the purpose of counteracting, alleviating, reducing, alleviating, ameliorating the condition of a disease or disorder such as sarcoma, etc.

Methods of treating kinase disorders

The invention also provides methods for treating disorders associated with abnormal mitogen extracellular kinase activity including, but not limited to, stroke, heart failure, hepatomegaly, cardiac enlargement, diabetes, alzheimer's disease, cystic fibrosis, symptoms of xenograft rejection, septic shock, or asthma.

An effective amount of a compound of the invention may be used to treat such disorders, including those diseases mentioned in the background section above (e.g., cancer). Moreover, such cancers and other diseases may be treated with the compounds of the present invention regardless of the mechanism of action and/or the relationship of the kinase to the condition.

The phrase "abnormal kinase activity" or "abnormal tyrosine kinase activity" includes any abnormal expression or activity of the gene encoding the kinase or the polypeptide encoded thereby. Examples of such aberrant activity include, but are not limited to, overexpression of the gene or polypeptide; gene amplification; mutations that produce constitutively active or highly active kinase activity; gene mutation, deletion, substitution, addition, and the like.

The present invention also provides methods of inhibiting kinase activity, particularly mitogen extracellular kinase activity, comprising administering an effective amount of a compound of the present invention, including salts, polymorphs, metabolites, hydrates, solvates, prodrugs (e.g., esters) and diastereomeric forms thereof. Kinase activity may be inhibited in cells (e.g., in vitro) or in cells of a mammalian subject, particularly a human patient in need of treatment.

Methods of treating angiogenic disorders

The invention also provides methods of treating conditions and diseases associated with excessive and/or abnormal angiogenesis.

Inappropriate and abnormal expression of angiogenesis can be harmful to an organism. Many pathological states are associated with the growth of new (extra) blood vessels. These include, for example, diabetic retinopathy, ischemic retinal vein occlusion, and retinopathy of prematurity [ Aiello et al, New engl.j.med.1994,331, 1480; peer et al, lab. invest.1995,72,638], age-related macular degeneration [ AMD; see Lopez et al invest, opthalmols, vis, sci, 1996,37,855], neovascular glaucoma, psoriasis, retrocrystallic fibroplasia, angiofibroma, inflammation, Rheumatoid Arthritis (RA), restenosis, in-stent restenosis, restenosis following vascular grafts, and the like. In addition, the increased blood supply associated with cancerous and tumor tissue promotes growth, resulting in rapid tumor enlargement and metastasis. In addition, the growth of new blood and lymph vessels in tumors provides an exit route for cancerous cells (renegade cells), promoting metastasis and leading to the spread of cancer. Thus, the compounds of the present invention may be used to treat and/or prevent any of the aforementioned angiogenic disorders, for example by inhibiting and/or reducing angiogenesis; inhibit, block, reduce, etc., endothelial cell proliferation or other types associated with angiogenesis, and cause cell death or apoptosis of such cells.

Dosage and administration

Effective dosages of the compounds of the present invention for the treatment of each of the desired indications can be readily determined based on standard laboratory techniques known to evaluate compounds for the treatment of hyperproliferative and angiogenic disorders, by standard toxicity tests, as well as by standard pharmacological tests for determining treatment of the disorders described hereinabove in mammals, and by comparing these results with those of known drugs used to treat these disorders. The amount of active ingredient administered in the treatment of one of these conditions may vary widely depending on the following considerations: the particular compound and dosage unit employed, the mode of administration, the course of treatment, the age and sex of the patient to be treated, and the nature and extent of the condition being treated.

The total amount of active ingredient to be administered is generally from about 0.001mg/kg to about 200mg/kg body weight/day, and preferably from about 0.01mg/kg to about 20mg/kg body weight/day. A clinically useful dosing regimen is one to three times daily dosing to once every four weeks dosing. In addition, a "drug withdrawal period" (where no drug is administered to the patient for a certain period of time) may be advantageous for the overall balance between pharmacological efficacy and tolerability. A unit dose may contain from about 0.5mg to about 2000mg of the active ingredient and may be administered one or more times per day, or less than once per day. The average daily dose administered by injection, including intravenous, intramuscular, subcutaneous and parenteral injection, and using infusion techniques, may preferably be from 0.01 to 200mg/kg of total body weight. The average daily rectal dosage regimen is preferably from 0.01 to 200mg/kg of total body weight. The average daily vaginal dosage regimen is preferably 0.01-200mg/kg total body weight. The average daily topical dosage regimen is preferably 0.1-200mg administered one to four times daily. The transdermal concentration is preferably the concentration required to maintain a daily dose of 0.01-200 mg/kg. The average daily inhaled dose regimen is preferably from 0.01 to 100mg/kg of total body weight.

The specific starting and maintenance dosage regimen for each patient will, of course, vary depending upon the following factors: the nature and severity of the condition as determined by the clinician, the activity of the particular compound used, the age and general health of the patient, the time of administration, the route of administration, the rate of excretion of the drug, the drug combination, and the like. The desired mode of treatment and the number of doses of a compound of the invention, or a pharmaceutically acceptable salt, or ester, or composition thereof, can be determined by one skilled in the art using routine therapeutic testing.

Preferably, the disease to which the method is directed is a hematological tumor, a solid tumor and/or metastases thereof.

The compounds of the invention are particularly useful in the treatment and prevention (i.e. prevention) of tumor growth and metastasis, particularly of solid tumors of all indications and stages, with or without prior treatment of said tumor growth.

Methods for determining specific pharmacological or pharmaceutical properties are well known to those skilled in the art.

The example assay experiments described herein are intended to exemplify the invention and the invention is not limited to the examples provided.

And (3) biological determination:

the examples were tested one or more times in selected bioassays. When tested more than once, the data is reported as a mean or median value, where:

the mean, also called arithmetic mean, represents the sum of the values obtained divided by the number of tests, and

the median value represents the number of the median position of the group of numerical values when arranged in ascending or descending order. If the number of values in the data set is odd, the median value is the middle value. If the number of values in the data set is even, the median value is the arithmetic mean of the two intermediate values.

The examples were synthesized one or more times. When synthesized more than once, the data from the bioassay represents mean or median values calculated using a data set obtained by testing one or more synthetic batches.

MKNK1 kinase assay

MKNK 1-inhibitory activity of the compounds of the invention was quantified using the MKNK1TR-FRET assay as described in the following paragraphs.

Recombinant fusion proteins (product No. 02-145) of glutathione-S-transferase (GST, N-terminally bound) and human full-length MKNK1 (amino acids 1-424 and T344D of accession No. BAA 19885.1), expressed in insect cells using a baculovirus expression system and purified by glutathione agarose affinity chromatography, were purchased from Carna Biosciences and used as enzymes. The biotinylated peptide biotin-Ahx-IKKRKLTRRKSLKG (C-terminus in the form of an amide) was used as a substrate for kinase reactions, which are available, for example, from Biosyntan (Berlin-Buch, Germany).

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ l of MKNK1 solution in aqueous assay buffer [50mM HEPES pH7.5, 5mM magnesium chloride, 1.0mM dithiothreitol, 0.005% (v/v) Nonidet-P40(Sigma) ] was added, and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding a solution of 3 μ L of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ L assay volume) and substrate (0.1 μ M ═ 0.06 μ M final concentration in 5 μ L assay volume) in assay buffer and the resulting mixture was incubated at 22 ℃ for a reaction time of 45 min. The concentration of MKNK1 was adjusted according to the activity of the enzyme batch and was chosen appropriately to bring the assay in the linear range, typical concentrations were in the range of 0.05 μ g/ml. The reaction was stopped by the addition of 5. mu.L of TR-FRET detection reagent solution (5nM streptavidin-XL 665[ Cisbio Bioassays, Codolet, France ] and 1nM anti-ribosomal protein S6(pSer236) -antibody [ #44921G ] from Invitrogen and 1nM solution of LANCE EU-W1024-labeled protein G [ Perkin-Elmer, product AD0071] in aqueous EDTA (100mM EDTA, 0.1% (W/v) bovine serum albumin in 50mM HEPES pH 7.5).

The resulting mixture was incubated at 22 ℃ for 1h to allow the formation of a complex between the phosphorylated biotinylated peptide and the detection reagent. The amount of phosphorylated substrate is then assessed by measuring the resonance energy transfer from the Eu-chelate to the streptavidin-XL. Thus, fluorescence emissions at 620nm and 665nm after excitation at 350nm are measured using a TR-FRET reader, such as Rubystar (BMGLABTtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 11 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values, the concentration range is 20. mu.M to 0.1nM (20. mu.M, 5.9. mu.M, 1.7. mu.M, 0.51. mu.M, 0.15. mu.M, 44nM, 13nM, 3.8nM, 1.1nM, 0.33nM and 0.1nM, the dilution series being prepared separately by serial dilution at 100-fold the level of concentrated DMSO solution prior to assay by 1: 3.4).

Table 1: MKNK1 IC₅₀

MKNK1 kinase high ATP assay

MKNK 1-inhibitory activity at high ATP of the compounds of the invention after their pre-incubation with MKNK1 was quantified using the TR-FRET based MKNK1 high ATP assay as described in the following paragraphs.

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ l of MKNK1 solution in aqueous assay buffer [50mM HEPES pH7.5, 5mM magnesium chloride, 1.0mM dithiothreitol, 0.005% (v/v) Nonidet-P40(Sigma) ] was added, and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding 3 μ L of a solution of adenosine triphosphate (ATP,3.3mM ═ 2mM final concentration in 5 μ L assay volume) and substrate (0.1 μ M ═ 0.06 μ M final concentration in 5 μ L assay volume) in assay buffer and the resulting mixture was incubated at 22 ℃ for a reaction time of 30 min. The concentration of MKNK1 was adjusted according to the activity of the enzyme batch and was chosen appropriately to bring the assay in the linear range, typical concentrations were in the range of 0.003 μ g/mL. The reaction was stopped by the addition of 5. mu.L of TR-FRET detection reagent solution (5nM streptavidin-XL 665[ Cisbio Bioassays, Codolet, France ] and 1nM anti-ribosomal protein S6(pSer236) -antibody [ #44921G ] from Invitrogen and 1nMLANCE EU-W1024-labeled protein G [ Perkin-Elmer, product AD0071] in aqueous EDTA (100mM EDTA, 0.1% (W/v) bovine serum albumin in 50mM HEPES pH 7.5).

The resulting mixture was incubated at 22 ℃ for 1h to allow the formation of a complex between the phosphorylated biotinylated peptide and the detection reagent. The amount of phosphorylated substrate is then assessed by measuring the resonance energy transfer from the Eu-chelate to the streptavidin-XL. Thus, fluorescence emissions at 620nm and 665nm after excitation at 350nm are measured using a TR-FRET reader, such as Rubystar (BMGLABTtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 11 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values in the range of 20. mu.M to 0.1nM (e.g., 20. mu.M, 5.9. mu.M, 1.7. mu.M, 0.51. mu.M, 0.15. mu.M, 44nM, 13nM, 3.8nM, 1.1nM, 0.33nM and 0.1nM, the dilution series being prepared separately by serial dilution at 100-fold the level of concentrated DMSO solution prior to assay, the exact concentration being variable depending on the pipette used).

Table 2: MKNK1 high ATP IC₅₀

CDK2/CycE kinase assay

CDK2/CycE inhibitory activity of compounds of the invention was quantified using the CDK2/CycE TR-FRET assay as described in the following paragraphs.

Recombinant fusion proteins of GST and human CDK2 and of GST and human CycE, which were expressed in insect cells (Sf9) and purified by glutathione-agarose affinity chromatography, were purchased from ProQinase GmbH (Freiburg, Germany). The biotinylated peptide biotin-Ttds-YISPLKSPYKISEG (C-terminus in amide form) was used as a substrate for kinase reactions, available, for example, from JERINI peptide science (Berlin, Germany).

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ l of CDK2/CycE was added in aqueous assay buffer [50mM Tris/HCl pH 8.0, 10mM magnesium chloride, 1.0mM dithiothreitol, 0.1mM sodium n-vanadate, 0.01% (v/v) Nonidet-P40(Sigma) ], and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding a solution of 3 μ l of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ l assay volume) and substrate (1.25 μ M ═ 0.75 μ M final concentration in 5 μ l assay volume) in assay buffer and the resulting mixture was incubated at 22 ℃ for a reaction time of 25 min. The concentration of CDK2/CycE was adjusted according to the activity of the enzyme batches and was chosen appropriately to bring the assay in a linear range, with a typical concentration in the range of 130 ng/mL. The reaction was stopped by the addition of 5. mu.L of TR-FRET detection reagent solution (0.2. mu.M streptavidin-XL 665[ Cisbio Bioassays, Codolet, France ] and 1nM anti-RB from BD Pharmingen (pSer807/pSer811) -antibody [ #558389] and 1.2nM LANCE EU-W1024 labeled anti-mouse IgG antibody [ Perkin-Elmer, product number AD0077, alternatively terbium-cryptate-labeled anti-mouse IgG antibody from Cisbio Bioassays ] in aqueous EDTA solution (100mM EDTA, 0.2% (W/v) bovine serum albumin in 100mM HEPES/NaOH pH 7.0).

The resulting mixture was incubated at 22 ℃ for 1h to allow the formation of a complex between the phosphorylated biotinylated peptide and the detection reagent. The amount of phosphorylated substrate is then assessed by measuring the resonance energy transfer from the Eu-chelate to the streptavidin-XL. Thus, fluorescence emissions at 620nm and 665nm after excitation at 350nm are measured using a TR-FRET reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 11 different concentrations, two values at each concentration, on the same microtiter plate, and IC50 values are calculated by 4-parameter fitting, ranging from 20 μ M to 0.1nM (20 μ M, 5.9 μ M, 1.7 μ M, 0.51 μ M, 0.15 μ M, 44nM, 13nM, 3.8nM, 1.1nM, 0.33nM, and 0.1nM, with the dilution series being prepared separately by 1:3.4 serial dilutions at the level of 100-fold concentrated DMSO solution prior to assay).

PDGFR beta kinase assay

PDGFR β inhibitory activity of the compounds of the invention was quantified using the PDGFR β HTRF assay as described in the following paragraphs.

As kinase, GST-His fusion protein containing the C-terminal fragment of human PDGFR β (amino acids 561-. Biotinylated poly-Glu, Tyr (4:1) copolymer (#61GT0BLA) from Cis biointentation (Marcoule, France) was used as a substrate for the kinase reaction.

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ l of PDGFR β was added to a solution in aqueous assay buffer [50mM HEPES/NaOH ph7.5, 10mM magnesium chloride, 2.5mM dithiothreitol, 0.01% (v/v) Triton-X100(Sigma) ], and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding 3 μ l of a solution of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ l assay volume) and substrate (2.27 μ g/mL ═ 1.36 μ g/mL [ about 30nM ] final concentration in 5 μ l assay volume) in assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 25 min. The PDGFR β concentration in the assay was adjusted according to the enzyme batch activity and was chosen appropriately to bring the assay in the linear range, with typical enzyme concentrations being in the range of about 125pg/μ L (final concentration in a5 μ L assay volume). The reaction was stopped by adding 5. mu.L of HTRF detection reagent solution (200nM streptavidin-XLent [ Cis BioInternational ] and 1.4nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Eu-chelate can also be replaced by PT 66-Tb-cryptate from Cis BioInternational ]) in aqueous EDTA solution (100mM EDTA, 0.2% (w/v) in 50mM HEPES/bovine serum albumin in NaOH pH 7.5).

The resulting mixture was incubated at 22 ℃ for 1h to bind biotinylated phosphorylated peptide to streptavidin-XLent and PT 66-Eu-chelate. The amount of phosphorylated substrate was subsequently evaluated by measuring the resonance energy transfer from PT 66-Eu-chelate to streptavidin-XLent. Thus, fluorescence emissions at 620nm and 665nm after excitation at 350nm were measured using an HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 10 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values, the concentration range from 20. mu.M to 1nM (20. mu.M, 6.7. mu.M, 2.2. mu.M, 0.74. mu.M, 0.25. mu.M, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, the dilution series being prepared by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

Fyn kinase assay

Human recombinant kinase domain with His 6-tag at the C-terminus of human T-Fyn (purchased from Invitrogen, P3042) was used as kinase, which was expressed in baculovirus-infected insect cells. The biotinylated peptide biotin-KVEKIGEGTYGVV (C-terminal in the form of an amide) is used as a substrate for kinase reactions, which are available, for example, from Biosynthan GmbH (Berlin-Buch, Germany).

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ L of a solution of T-Fyn in aqueous assay buffer [25mM Tris/HCl ph7.2, 25mM magnesium chloride, 2mM dithiothreitol, 0.1% (w/v) bovine serum albumin, 0.03% (v/v) Nonidet-P40(Sigma) ], and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme prior to starting the kinase reaction. Then, the kinase reaction was started by adding a solution of 3 μ l of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ l assay volume) and substrate (2 μ M ═ 1.2 μ M final concentration in 5 μ l assay volume) in assay buffer and the resulting mixture was incubated at 22 ℃ for a reaction time of 60 min. The concentration of Fyn is adjusted according to the activity of the enzyme batch and is chosen appropriately to bring the assay in the linear range, typical concentration is 0.13 nM. The reaction was stopped by adding 5. mu.L of HTRF detection reagent solution (0.2. mu.M solution of streptavidin-XL [ Cisbio Bioassays, Codolet, France ] and 0.66nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Tb-chelate from Cisbio Bioassays can also be used instead of PT 66-Eu-chelate ]) in aqueous EDTA solution (125mM EDTA, 0.2% (w/v) bovine serum albumin pH7.0 in 50mM HEPES/NaOH)).

The resulting mixture was incubated at 22 ℃ for 1h to bind the biotinylated phosphorylated peptides to the streptavidin-XL and PT 66-Eu-chelate. The amount of phosphorylated substrate was subsequently assessed by measuring the resonance energy transfer from PT 66-Eu-chelate to streptavidin-XL. Thus, HTRF is utilizedA reader, for example a Rubystar (BMG Labtechnologies, Offenburg, Germany) or a Viewlux (Perkin-Elmer) measures the fluorescence emission at 620nm and 665nm after excitation at 350 nm. The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 10 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values, the concentration range from 20. mu.M to 1nM (20. mu.M, 6.7. mu.M, 2.2. mu.M, 0.74. mu.M, 0.25. mu.M, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, the dilution series being prepared by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

Flt4 kinase assay

Flt4TR-FRET assays as described in the following paragraphs were used to quantify the Flt4 inhibitory activity of the compounds of the invention.

As kinase, GST-His fusion proteins containing the C-terminal fragment of human Flt4 (amino acids 799-1298), purchased from Proqinase [ Freiburg i.Brsg., Germany ], expressed in insect cells [ SF9] and purified by affinity chromatography were used. The biotinylated peptide biotin-Ahx-GGEEEEYFELVKKKK (C-terminus in amide form, available from Biosyntan, Berlin-Buch, Germany) was used as a substrate for the kinase reaction.

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ L of Flt4 in aqueous assay buffer [25mM HEPES ph7.5, 10mM magnesium chloride, 2mM dithiothreitol, 0.01% (v/v) Triton-X100(Sigma), 0.5mM EGTA, and 5mM β -glycerophosphate ], and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding 3 μ L of a solution of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ L assay volume) and substrate (1.67 μ M ═ 1 μ M final concentration in 5 μ L assay volume) in assay buffer and the resulting mixture was incubated at 22 ℃ for a reaction time of 45 min. The Flt4 concentration in the assay was adjusted according to the activity of the enzyme batch and was chosen appropriately to bring the assay in the linear range, with typical enzyme concentrations being in the range of about 120 pg/. mu.L (final concentration in a 5. mu.l assay volume). The reaction was stopped by adding 5 μ L of HTRF detection reagent solution (200nM streptavidin-XL 665[ Cis Biointernational ] and 1nM PT 66-Tb-cryptate (terbium-cryptate labeled anti-phosphotyrosine antibody from Cisbio Bioassays (Codolet, France)) in aqueous EDTA (50mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES pH 7.5).

The resulting mixture was incubated at 22 ℃ for 1h to allow binding of biotinylated phosphorylated peptides to streptavidin-XL 665 and PT 66-Tb-cryptate. The amount of phosphorylated substrate was then assessed by measuring the resonance energy transfer from PT 66-Tb-cryptate to streptavidin-XL 665. Thus, fluorescence emissions at 620nm and 665nm after excitation at 350nm were measured using an HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 10 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values, the concentration range from 20. mu.M to 1nM (20. mu.M, 6.7. mu.M, 2.2. mu.M, 0.74. mu.M, 0.25. mu.M, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, the dilution series being prepared by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

TrkA kinase assay

TrkA inhibitory activity of the compounds of the present invention was quantified using the TrkA HTRF assay as described in the following paragraphs.

As kinase, GST-His fusion protein containing the C-terminal fragment of human TrkA (amino acid 443-796) purchased from Proqinase [ Freiburg i.Brsg., Germany ], which was expressed in insect cells [ SF9] and purified by affinity chromatography was used. Biotinylated poly-Glu, Tyr (4:1) copolymer (#61GT0BLA) from Cis biointentation (Marcoule, France) was used as a substrate for the kinase reaction.

For the assay, 50nL of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black low-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 μ L of a solution of TrkA in aqueous assay buffer [8mM MOPS/HCl ph7.0, 10mM magnesium chloride, 1mM dithiothreitol, 0.01% (v/v) NP-40(Sigma), 0.2mM EDTA ], and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme before starting the kinase reaction. Then, the kinase reaction was started by adding 3 μ L of a solution of adenosine triphosphate (ATP,16.7 μ M ═ 10 μ M final concentration in 5 μ L assay volume) and substrate (2.27 μ g/mL ═ 1.36 μ g/mL [ about 30nM ] final concentration in 5 μ L assay volume) in assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 60 min. The TrkA concentration in the assay is adjusted according to the activity of the enzyme batch and is chosen appropriately so that the assay is in the linear range, typical enzyme concentrations are in the range of about 20pg/μ L (final concentration in a5 μ L assay volume). The reaction was stopped by adding 5. mu.L of HTRF detection reagent solution (30nM streptavidin-XL 665[ Cis Biointernational ] and 1.4nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Eu-chelate can also be replaced by PT 66-Tb-cryptate from Cis Biointernational ]) in aqueous EDTA solution (100mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES/pH 7.5).

The resulting mixture was incubated at 22 ℃ for 1h to bind biotinylated phosphorylated peptides to streptavidin-XL 665 and PT 66-Eu-chelate. The amount of phosphorylated substrate was subsequently assessed by measuring the resonance energy transfer from PT 66-Eu-chelate to streptavidin-XL 665. Thus, fluorescence at 620nm and 665nm after excitation at 350nm is measured using an HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer)And (4) transmitting. The ratio of the emissions at 665nm and 622nm was used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor 0% inhibition with all other assay components without enzyme 100% inhibition). Typically, test compounds are tested at 10 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated by 4-parameter fitting₅₀Values, the concentration range from 20. mu.M to 1nM (20. mu.M, 6.7. mu.M, 2.2. mu.M, 0.74. mu.M, 0.25. mu.M, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, the dilution series being prepared by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

AlphaScreen Surefire eIF4E Ser209 phosphorylation assay

The AlphaScreen SureFire eIF4E Ser209 phosphorylation assay was used to measure the phosphorylation of endogenous eIF4E in cell lysates. The AlphaScreen SureFire technology allows for the detection of phosphorylated proteins in cell lysates. In this assay, sandwich antibody complexes formed only in the presence of the analyte (p-eIF4E Ser209) were captured in close proximity by AlphaScreen donor and acceptor microbeads. Excitation of the donor bead causes the release of singlet oxygen molecules, which trigger an energy transfer cascade in the acceptor bead, producing light emission at 520-620 nm.

Surefire EIF4e Alphascreen stimulated with 20% FCS in A549 cells

For the assay, the AlphaScreen SureFire p-eIF4E Ser20910K assay kit and AlphaScreen protein A kit (for 10K assay points), both from Perkin Elmer, were used.

On the first day, 50,000 a549 cells were seeded in 96-well plates at 100 μ L per well in growth medium (DMEM/Hams' F12 with stabilized glutamine, 10% FCS) and incubated at 37 ℃. After cell attachment, the medium was changed to starvation medium (DMEM, 0.1% FCS, no glucose, with glutamine, supplemented with 5g/L maltose). On the next day, test compounds were added in DMSO and to a549 cells in the test plate at final concentrations ranging from up to 30 μ M to down to 10nM depending on the activity of the test compound. The treated cells were incubated at 37 ℃ for 2 hours. FCS was added to the wells over 20 min to a final FCS concentration of 20%. The medium was then removed and the cells were lysed by adding 50 μ Ι _ of cell lysis buffer. Then, the plate was shaken on a plate shaker for 10 min. After 10min cell lysis time, 4 μ L of lysate was transferred to 384 well plates (Proxiplate, from Perkin Elmer) and 5 μ L of reaction buffer containing AlphaScreen receptor microbeads plus activation buffer mix was added. Plates were sealed with TopSeal-A adhesive film and gently shaken on a plate shaker at room temperature for 2 h. Thereafter, 2 μ L of dilution buffer with AlphaScreen donor beads was added under soft light and plates were sealed again with TopSeal-a glue film and covered with foil. Incubate for another 2h and shake gently at room temperature. The plates were then measured in an EnVision reader (Perkin Elmer) with AlphaScreen program. Each data point (compound dilution) was measured in triplicate.

Determination of IC by 4-parameter fitting₅₀The value is obtained.

Assays for other MKNK-1 kinases can be similarly performed using suitable reagents, as will be apparent to those skilled in the art.

Accordingly, the compounds of the present invention are effective in inhibiting one or more MKNK-1 kinases and are therefore suitable for treating or preventing diseases caused by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response, or an inappropriate cellular inflammatory response, particularly wherein said uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response, or inappropriate cellular inflammatory response is mediated by MKNK-1, more particularly wherein said diseases caused by uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response, or inappropriate cellular inflammatory response are hematological tumors, solid tumors, and/or their metastases, such as leukemias and myelodysplastic syndromes, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, tumors, and metastases, Breast, gastrointestinal, endocrine, breast and other gynaecological tumours including non-small cell and small cell lung tumours, urological tumours including renal, bladder and prostate tumours, skin tumours and sarcomas, and/or metastases thereof.

Claims

1. A compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same:

wherein:

r1 represents C₂-C₆-alkyl-or C₃-C₆Cycloalkyl radicals, to which they are attachedGroup a, and they are optionally substituted once, twice or three times independently of each other by a substituent selected from:

wherein the group a represents a substituent selected from:

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

r2 represents a substituent selected from:

hydrogen atom or C₁-C₆-an alkyl group;

r represents a substituent selected from:

r' and R "independently of each other represent a substituent selected from:

C₁-C₆-alkyl-, C₃-C₆-cycloalkyl-, C₁-C₆-haloalkyl-, C₁-C₆-alkoxy-C₂-C₆-alkyl-or C₁-C₆-haloalkoxy-C₂-C₆-an alkyl-group.

2. A compound according to claim 1, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

r1 represents C₂-C₆-alkyl-or C₃-C₆-cycloalkyl radicalThey are substituted by a group a, and they are optionally substituted once, twice or three times, independently of one another, by a substituent selected from the group consisting of:

wherein the group a represents a substituent selected from:

--NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)R’、-N(R’)C(＝O)R’、-N(H)C(＝O)NH₂、-N(H)C(＝O)NHR’、-N(H)C(＝O)N(R’)R”、-N(R’)C(＝O)NH₂、-N(R’)C(＝O)NHR’、-N(R’)C(＝O)N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂-OC (═ O) NHR ', or-OC (═ O) N (R') R "groups,

or:

-azetidinyl, or 5-to 7-membered nitrogen-containing heterocycloalkyl,

r2 represents a substituent selected from:

hydrogen atom or C₁-C₃-an alkyl group;

r' and R "independently of each other represent a substituent selected from:

C₁-C₃-alkyl-, C₃-C₄-cycloalkyl-, C₁-C₃-haloalkyl-, C₁-C₃-alkoxy-C₂-C₃-alkyl-or C₁-C₃-haloalkoxy-C₂-C₃-an alkyl-group.

3. A compound according to claim 1 or2, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

r1 represents C₂-C₆-alkyl, substituted by a group a, and optionally substituted by C₁-C₃-alkyl is substituted once, twice or three times independently of each other;

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group,

or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

-said 5-to 7-membered nitrogen-containing heterocycloalkyl optionally substituted by C₁-C₃-alkyl is substituted once or twice independently of each other;

r2 represents a substituent selected from:

a hydrogen atom, or C₁-C₃-an alkyl group;

r 'and R' independently of one another represent C₁-C₃-an alkyl group.

4. A compound according to any one of claims 1 to 3, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

wherein the group a represents a substituent selected from:

--NH₂n (R ') R ", -N (h) C (═ O) R', -OH, or C₁-C₃-an alkoxy group,

or:

-a 5-to 7-membered nitrogen-containing heterocycloalkyl group,

r2 represents a substituent selected from:

hydrogen atom or C₁-C₃-an alkyl group;

r 'and R' independently of one another represent C₁-C₃-an alkyl group.

5. The compound according to any one of claims 1 to 4, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

wherein the group a represents a substituent selected from:

--NH₂-N (R ') R ", -N (H) C (═ O) R', -OH or methoxy,

or:

-a 5-or 6-membered nitrogen-containing heterocycloalkyl group,

-said 5-or 6-membered nitrogen-containing heterocycloalkyl optionally substituted with a methyl group;

r2 represents a substituent selected from:

a hydrogen atom or a methyl group;

r 'and R' independently of one another represent a methyl or ethyl group.

6. A compound according to any one of claims 1 to 5, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, selected from the group consisting of:

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-ol;

3- (6-methoxy-1-benzofuran-2-yl) -6- (3-methoxypropoxy) imidazo [1,2-b ] pyridazine;

2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } ethanamine;

2- (6- { [ (2R) -1-aminopropan-2-yl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) -1-benzofuran-6-ol;

n- [ (2R) -2- { [3- (6-hydroxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide;

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine;

3- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -N, 2, 2-tetramethylpropan-1-amine;

4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } butan-1-amine;

4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -N, N-dimethylbut-1-amine;

n, N-diethyl-4- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } pentan-1-amine;

3- (6-methoxy-1-benzofuran-2-yl) -6- [2- (1-methylpyrrolidin-2-yl) ethoxy ] imidazo [1,2-b ] pyridazine;

1- (2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } ethyl) imidazolidin-2-one;

3- (6-methoxy-1-benzofuran-2-yl) -6- [3- (morpholin-4-yl) propoxy ] imidazo [1,2-b ] pyridazine;

n- [ (2R) -2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propyl ] acetamide; and

(2R) -2- { [3- (6-methoxy-1-benzofuran-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine.

7. A compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, according to any one of claims 1 to 6, for use in the treatment or prophylaxis of a disease.

8. A pharmaceutical composition comprising a compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, according to any one of claims 1 to 6, and a pharmaceutically acceptable diluent or carrier.

9. A pharmaceutical combination comprising:

-one or more first active ingredients selected from compounds of general formula (I) according to any one of claims 1 to 6, and

-one or more second active ingredients selected from chemotherapeutic anti-cancer agents and target-specific anti-cancer agents.

10. Use of a compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, according to any one of claims 1 to 6, for the prophylaxis or treatment of a disease.

11. Use of a compound of general formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, or a mixture of same, according to any one of claims 1 to 6, for the preparation of a medicament for the prophylaxis or treatment of a disease.

12. Use according to claim 7, 10 or 11, wherein the disease is a disease caused by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response, in particular wherein the uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is mediated by the MKNK-1 pathway, more in particular wherein the disease caused by uncontrolled cell growth, proliferation and/or survival, inappropriate cellular immune response or inappropriate cellular inflammatory response is a hematological tumor, a solid tumor and/or their metastases, such as leukemia and myelodysplastic syndrome, malignant lymphoma, head and neck tumors including brain tumors and brain metastases, breast tumors including non-small cell lung tumors and small cell lung tumors, breast tumors including non-small cell lung tumors, brain metastases, brain tumors, brain, Gastrointestinal tumors, endocrine tumors, breast tumors and other gynecological tumors, urological tumors including renal tumors, bladder tumors and the foregoing adenomas, skin tumors and sarcomas, and/or metastases thereof.

13. A compound selected from:

wherein R2 is as defined for a compound of general formula (I) in any one of claims 1 to 6, wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, such as a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group;

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R7 represents a protecting group, such as a silyl protecting group, for example, a tert-butyldimethylsilyl group;

wherein R1 is as defined for compounds of general formula (I) in claims 1 to 6, and wherein R7 represents a protecting group, such as a silyl protecting group, e.g. tert-butyldimethylsilyl; and

14. Use of a compound selected from the group consisting of,

wherein X and Y represent a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group;

wherein R2 is as defined for the compounds of general formula (I) in claims 1 to 6, wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group;

wherein R1 is as defined for the compound of general formula (I) in claims 1 to 6, and wherein Y represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group;

wherein X represents a leaving group, such as a halogen atom, for example a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group, for example a trifluoromethylsulfonate group, a nonafluorobutylsulfonate group, and wherein R7 represents a protecting group, for example, a silyl protecting group, for example, a tert-butyldimethylsilyl group;

wherein R1 is as defined for compounds of general formula (I) in claims 1 to 6, and wherein R7 represents a protecting group, e.g. a silyl protecting group, e.g. tert-butyldimethylsilyl; and