HK1203195B

HK1203195B - Amino-substituted imidazopyridazines

Info

Publication number: HK1203195B
Application number: HK15103714.2A
Authority: HK
Inventors: K.艾斯; F.普勒; L.措恩; V.舒尔策; D．苏兹勒; P.利瑙; A．哈格巴尔特; K.彼得森; U.伯默
Original assignee: Bayer Intellectual Property Gmbh
Priority date: 2011-12-12
Filing date: 2012-12-10
Publication date: 2017-12-01

Description

Amino-substituted imidazopyridazines

The present invention relates to 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) 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, structure13,1559-1568,2005 and Jauch et al, EMBO J25, 4020-. 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, whereas MKNK2b had basal activity independent of ERK or p38MAP kinase. [ Buxade M et al, Frontiers in Bioscience 5359-5374,2008 5 months and 1 day ]

MKNK has been shown to phosphorylate eukaryotic initiation factor 4E (eIF4E), heterogeneous nuclear RNA-binding protein A1(hnRNPA1), polypyrimidine-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 biol24,6539-6549,2004 ]. eIF4E plays a key role in 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 generally translated less efficiently 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 the like. 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 a "weak mRNA" 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 poor prognosis in patients with small cell lung Cancer [ 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 dead MKNK1) in mouse embryonic fibroblasts promoted tumor formation [ Chrestensen c.a. et al, Genes Cells 12, 1133-. Furthermore, increased MKNK phosphorylation and activity is associated with overexpression of HER2 in breast cancer [ 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, MKNK1, constitutively activated (but not kinase dead), promoted tumor growth. Comparable results were achieved when eIF4E with the S209D mutation was analyzed. The S209D mutation mimics phosphorylation at the MKNK1 phosphorylation site. In contrast, the non-phosphorylatable form of eIF4E attenuates 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.

WO 2007/025540 a2(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.

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

WO 2007/013673 a1(Astellas Pharma Inc.) relates to fused heterocycles as inhibitors of lymphocyte protein tyrosine kinases (abbreviated "LCK"). In particular, WO 2007/013673A 1 relates inter alia to imidazo [1,2-b ] pyridazines.

WO 2007/147646 a1(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.

WO 2008/025822 a1(cellzome (uk) Ltd.) relates to oxadiazodiazine derivatives for use as kinase inhibitors. In particular, WO 2008/025822 a1 relates inter alia to imidazo [1,2-b ] pyridazines useful as kinase inhibitors, in particular inducible T cell kinase (abbreviated as "Itk") inhibitors.

WO 2008/030579 a2(Biogen Idec MA Inc.) relates to modulators of interleukin-1 (IL-1) receptor associated kinase (abbreviated as "IRAK"). In particular, WO 2008/030579A 2 relates inter alia to imidazo [1,2-b ] pyridazines.

WO 2008/058126 a2(Supergen, Inc.) relates inter alia to imidazo [1,2-b ] pyridazine derivatives which act as protein kinase inhibitors, in particular PIM kinase inhibitors.

WO 2009/060197 a1(Centro Nacional de investigations Oncology (CNIO)) relates to imidazopyridazines for use as protein kinase inhibitors, such as PIM family kinases.

US 4,408,047(Merck & co., Inc.) relates inter alia to imidazopyridazines with 3-amino-2-OR-propoxy substituents, which have β -adrenergic blocking activity.

WO 03/018020A 1(Takeda Chemical Industries, Ltd.) relates to inhibitors against c-Jun N-terminal kinases comprising compounds which are inter alia imidazo [1,2-b ] pyridazine.

WO 2008/052734 a1(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.

WO 2008/072682 a1(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.

WO 2008/079880 a1(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.

WO 2009/091374 a2(Amgen Inc.) relates to fused heterocyclic derivatives. The selected compounds are effective in preventing and treating diseases, such as hepatocyte growth factor ("HGF") diseases.

An article entitled "Structural Basis of the Inhibitor Specificity of the protooncogenic viral infection Site in molecular tissue Virus (PIM-1) Kinase" is described in J.Med.chem.,2005,487604-7614, and in particular discloses imidazo [1,2-b ] as inhibitor structures for use in the studies described therein]Pyridazine.

Named "Discovery of MiAn article by togen-Activated Protein Kinase-Interacting Kinase1Inhibitors by a Comprehensive Fragment-organized visual Screening Approach "is described in j.med.chem.,2010,536618-6628, and especially in Table 1, disclose some specific imidazo [1,2-b ] compounds identified as MKNK-1 inhibitors]Pyridazine.

An article entitled "Therapeutic inhibition of MAP inhibition of blocking inhibition of binding of polypeptides" is described in Cancer Res, 3/1/2011,711849-1857, and in particular, discloses that the known antifungal agent Cercosporamide (Cercosporamide) is an inhibitor of MKNK 1.

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

-having the following group in its 3-position:

a group having the following structure at its 6-position:

wherein:

indicates the point of attachment of the group to the rest of the molecule,

-R1 represents an optionally substituted linear chain C as defined herein₂-C₆-alkyl-, straight chain C₃-C₆-alkyl-or C₃-C₆A cycloalkyl radical, and

-R2 represents a substituent as defined herein;

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

It has now been found that the compounds of the invention have surprising and advantageous properties and this forms 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 hematological tumors, solid tumors and/or metastases thereof, such as leukemia and myelodysplastic syndromes, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, head and neck tumors, and head and neck tumors including brain tumors and metastases, and/or metastases thereof, 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.

Disclosure of Invention

According to a first aspect, the present invention covers 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:

represents a group selected from:

wherein indicates the point of attachment of the group to the rest of the molecule;

r1 represents a straight-chain C optionally substituted by one or more substituents selected independently of one another from the group consisting of₂-C₆Alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-a cycloalkyl group:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 6-membered heterocycloalkyl linked as a spiro ring; aryl optionally substituted one or more times by R substituents, C₁-C₆-alkoxy-, heteroaryl optionally substituted one or more times by R substituents-, -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’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-an alkyl-S-group;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, -OC (═ O) R', -SH, C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂N (R ') R ", -S (═ O) (═ NR') R";

r4 represents a substituent selected from:

hydrogen atom, halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 10-membered heterocycloalkyl-, aryl optionally substituted one or more times by R substituents, heteroaryl optionally substituted one or more times by R substituents, C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-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 ", -S (═ O) (═ NR') R";

r represents a substituent selected from the group consisting of:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 10-membered heterocycloalkyl-, aryl-, heteroaryl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-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 ", -S (═ O) (═ NR') R";

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0,1, 2 or 3.

According to an embodiment of the first aspect, the present invention covers a compound of formula (I) as described above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein

Represents a group selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-; aryl-optionally substituted one or more times independently of each other by R substituents; aryl-C optionally substituted one or more times independently of each other by R substituents₁-C₆-alkoxy-; heteroaryl-optionally substituted one or more times independently of each other by R substituents; -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’、-OH、C₁-C₆-alkoxy-,C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-an alkyl-S-group;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -SH, C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂N (R ') R ", -S (═ O) (═ NR') R";

r4 represents a substituent selected from:

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0,1, 2 or 3.

The terms mentioned herein preferably have the following meanings:

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

The term "C₁-C₆Alkyl "is understood to preferably mean 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-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 isomers thereof. In particular, the radicals have 1,2, 3 or 4 carbon atoms ("C)₁-C₄-alkyl "), such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, more particularly said group having 1,2 or 3 carbon atoms (" C)₁-C₃-alkyl "), such as methyl, ethyl, n-propyl or isopropyl.

The term "halo-C₁-C₆Alkyl is understood to mean preferably a straight-chain or branched, saturated monovalent hydrocarbon radical, where the term "C" is₁-C₆Alkyl radicals "being as defined aboveAnd wherein one or more hydrogen atoms are replaced by halogen atoms in the same or different manner, i.e. independently of each other. In particular, the halogen atom is F. Said halo-C₁-C₆Alkyl is, for example, -CF₃、-CHF₂、-CH₂F、-CF₂CF₃or-CH₂CF₃。

The term "C₁-C₆Alkoxy is to be understood as preferably meaning a straight-chain or branched, saturated monovalent hydrocarbon radical of the formula-O-alkyl, where the term "alkyl" is defined as above, for example methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, pentoxy, isopentoxy or n-hexoxy, or isomers thereof. In particular, the said "C₁-C₆Alkoxy "may contain 1,2 or 3 carbon atoms (" C)₁-C₃-alkoxy ").

The term "halo-C₁-C₆Alkoxy is understood as preferably meaning a straight-chain or branched, saturated, monovalent C radical as defined above in which one or more hydrogen atoms are replaced by halogen atoms in the same or different manner₁-C₆-alkoxy groups. In particular, the halogen atom is F. Said halo-C₁-C₆Alkoxy is, for example, -OCF₃、-OCHF₂、-OCH₂F、-OCF₂CF₃or-OCH₂CF₃。

The term "C₁-C₆-alkoxy-C₁-C₆Alkyl is understood to preferably denote-C in which one or more hydrogen atoms are defined as above in the same or different manner₁-C₆Alkoxy substituted straight-chain or branched saturated monovalent alkyl radicals as defined above or their isomers, for example methoxyalkyl, ethoxyalkyl, propoxyalkyl, isopropoxyalkyl, butoxyalkyl, isobutoxyalkyl, tert-butoxyalkyl, sec-butoxyalkyl, pentoxyalkyl, isopentyloxyalkyl, hexyloxyalkyl, where the term "C" is used₁-C₆-alkyl "As defined above.

The term "halo-C₁-C₆-alkoxy-C₁-C₆Alkyl is understood to preferably denote straight-chain or branched, saturated, monovalent-C as defined above in which one or more hydrogen atoms are replaced by halogen atoms in the same or different manner₁-C₆-alkoxy-C₁-C₆-an alkyl group. In particular, the halogen atom is F. Said halo-C₁-C₆-alkoxy-C₁-C₆Alkyl is, for example, -CH₂CH₂OCF₃、-CH₂CH₂OCHF₂、-CH₂CH₂OCH₂F、-CH₂CH₂OCF₂CF₃or-CH₂CH₂OCH₂CF₃。

The term "C₂-C₆Alkenyl "is understood to preferably mean a straight-chain or branched, monovalent hydrocarbon radical which contains one or more double bonds and has 2,3,4, 5 or 6 carbon atoms, in particular 2 or 3 carbon atoms (" C)₂-C₃-alkenyl "), it being understood that in case the alkenyl group comprises more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl group is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, homoallyl, (E) -but-2-enyl, (Z) -but-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, m, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3-enyl, (E) -hex-2-enyl, (Z) -hex-2-enyl, (E) -hex-1-enyl, (Z) -hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2-enyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E) -2-methylbut-1-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl, 4-methylpent-4-enyl, 3-methylpent-4-enyl, 2-methylpent-4-enyl, 1-methylpent-4-enyl, 4-methylpent-3-enyl, (E) -3-methylpent-3-enyl, methyl-but-1-enyl, methyl-1-propenyl, methyl-but-4-enyl, methyl-pent-3, (Z) -3-methylpent-3-enyl, (E) -2-methylpent-3-enyl, (Z) -2-methylpent-3-enyl, (E) -1-methylpent-3-enyl, (Z) -1-methylpent-3-enyl, (E) -4-methylpent-2-enyl, (Z) -4-methylpent-2-enyl, (E) -3-methylpent-2-enyl, (Z) -3-methylpent-2-enyl, (E) -2-methylpent-2-enyl, (Z) -2-methylpent-2-enyl, (E) -1-methylpent-2-enyl, methyl-2-enyl, (Z) -1-methylpent-2-enyl, (E) -4-methylpent-1-enyl, (Z) -4-methylpent-1-enyl, (E) -3-methylpent-1-enyl, (Z) -3-methylpent-1-enyl, (E) -2-methylpent-1-enyl, (Z) -2-methylpent-1-enyl, (E) -1-methylpent-1-enyl, (Z) -1-methylpent-1-enyl, 3-ethylbut-3-enyl, 2-ethylbut-3-enyl, 1-ethylbut-3-enyl, methyl-1-pentenyl, ethyl-3-enyl, ethyl-2-pentenyl, ethyl-1-, (E) -3-ethylbut-2-enyl, (Z) -3-ethylbut-2-enyl, (E) -2-ethylbut-2-enyl, (Z) -2-ethylbut-2-enyl, (E) -1-ethylbut-2-enyl, (Z) -1-ethylbut-2-enyl, (E) -3-ethylbut-1-enyl, (Z) -3-ethylbut-1-enyl, 2-ethylbut-1-enyl, (E) -1-ethylbut-1-enyl, (Z) -1-ethylbut-1-enyl, 2-propylprop-2-enyl, 1-propylprop-2-enyl, 2-isopropylprop-2-enyl, 1-isopropylprop-2-enyl, (E) -2-propylprop-1-enyl, (Z) -2-propylprop-1-enyl, (E) -1-propylprop-1-enyl, (Z) -1-propylprop-1-enyl, (E) -2-isopropylprop-1-enyl, (Z) -2-isopropylprop-1-enyl, (E) -1-isopropylprop-1-enyl, (Z) -1-isopropylprop-1-enyl, (E) -3, 3-dimethylprop-1-enyl, and (Z) -1-isopropylprop-1-enyl), (Z) -3, 3-dimethylprop-1-enyl, 1- (1, 1-dimethylethyl) vinyl, but-1, 3-dienyl, penta-1, 4-dienyl, hex-1, 5-dienyl or methylhexadienyl. In particular, the group is vinyl or allyl.

The term "C₂-C₆Alkynyl is understood as preferably meaning a straight-chain or branched, monovalent hydrocarbon radical which comprises one or more triple bonds and comprisesContaining 2,3,4, 5 or 6 carbon atoms, especially 2 or 3 carbon atoms ("C)₂-C₃-alkynyl "). Said C is₂-C₆Alkynyl is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, prop-2-ynyl, but-3-methylbut-1-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2-dimethylbut-3-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-1-ynyl, 3-methylpent-1-, 1, 1-dimethylbut-3-ynyl, 1-dimethylbut-2-ynyl or 3, 3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

The term "C₃-C₁₀Cycloalkyl "is understood to mean a saturated, monovalent, monocyclic or bicyclic hydrocarbon ring comprising 3,4, 5,6, 7,8, 9 or 10 carbon atoms (" C)₃-C₁₀-cycloalkyl "). Said C is₃-C₁₀Cycloalkyl is, for example, a monocyclic hydrocarbon ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon ring such as a perhydropentalenene or decahydronaphthalene ring. In particular, the ring contains 3,4, 5 or 6 carbon atoms ("C)₃-C₆-cycloalkyl ").

The term "C₃-C₆-cycloalkoxy "is to be understood as preferably representing a saturated monovalent hydrocarbon ring containing 3,4, 5 or 6 carbon atoms of formula-O-cycloalkyl (wherein the term" cycloalkyl "is as defined above), such as for example cyclopropyloxy, cyclobutyloxy, cyclopentyloxy or cyclohexyloxy.

The term "C₃-C₆-cycloalkyl-C₁-C₃Alkoxy is understood to mean preferably C in which one hydrogen atom is as defined above₃-C₆-saturated monovalent alkoxy groups as defined above, substituted by cycloalkyl groups, such as cyclopropylalkoxy, cyclobutylalkoxy, cyclopentylalkoxy, cyclohexylalkoxy groups (wherein the term "alkoxy" is as defined above) or isomers thereof.

The term "C₄-C₁₀Cycloalkenyl is understood as preferably meaning a monovalent monocyclic or bicyclic hydrocarbon ring which contains 4,5, 6, 7,8, 9 or 10 carbon atoms and 1,2, 3 or 4 conjugated or unconjugated double bonds, provided that the size of the cycloalkenyl ring allows this. For example, the C₄-C₁₀Cycloalkenyl is a monocyclic hydrocarbon ring such as cyclobutenyl, cyclopentenyl or cyclohexenyl, or a bicyclic hydrocarbon ring, for example:

the term "3-to 10-membered heterocycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 2,3,4, 5,6, 7,8 or 9 carbon atoms and one or more groups selected from C (═ O), O, S, S (═ O), S (═ O)₂、NR^aWherein R is^aRepresents a hydrogen atom, C₁-C₆-alkyl-or halo-C₁-C₆-an alkyl-group; the heterocycloalkyl group may be attached to the rest of the molecule through any of the carbon atoms or the nitrogen atom (if present).

In particular, the 3-to 10-membered heterocycloalkyl group can comprise 2,3,4, or 5 carbon atoms and one or more of the above-described heteroatom-containing groups ("3-to 6-membered heterocycloalkyl"), more particularly, the heterocycloalkyl group can comprise 4 or 5 carbon atoms and one or more of the above-described heteroatom-containing groups ("5-to 6-membered heterocycloalkyl").

In particular, the heterocycloalkyl group may be, for example but not limited to: 4-membered rings such as azetidinyl, glycidylalkyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or a 7-membered ring, such as a diazepanyl ring. Optionally, the heterocycloalkyl group can be benzofused.

The heterocyclyl group may be bicyclic, such as, but not limited to, a5, 5-membered ring, for example, a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a5, 6-membered bicyclic ring, for example, a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring.

As noted above, the nitrogen atom containing ring may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to a2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydrooxazolyl or 4H- [1,4] thiazinyl ring, or it may be benzo-fused, such as but not limited to a dihydroisoquinolinyl ring.

The term "4-to 10-membered heterocycloalkenyl" is understood to mean an unsaturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 3,4, 5,6, 7,8 or 9 carbon atoms and one or more groups selected from C (═ O), O, S, S (═ O), S (═ O)₂、NR^aWherein R is^aRepresents a hydrogen atom, C₁-C₆-alkyl-or halo-C₁-C₆-an alkyl-group; the heterocycloalkenyl group may be attached to the rest of the molecule through any of the carbon atoms or the nitrogen atom (if present). Examples of said heterocycloalkenyl may comprise one or more double bonds, such as 4H-pyranyl, 2H-pyranyl, 3H-bisaziridinyl, 2, 5-dihydro-1H-pyrrolyl, [1,3 ] -l]Dioxolyl ([1,3 ])]dioxolyl)、4H-[1,3,4]Thiadiazinyl, 2, 5-dihydrofuryl, 2, 3-dihydrofuryl, 2, 5-dihydrothienyl, 2, 3-dihydrothienyl, 4, 5-dihydrooxazolyl or 4H- [1,4 [ ]]Thiazinyl, or it may be benzo-fused.

The term "aryl" is to be understood as meaningPreferably represents a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6, 7,8, 9, 10, 11, 12, 13 or 14 carbon atoms ("C₆-C₁₄Aryl "), in particular a ring having 6 carbon atoms (" C)₆-aryl ") such as phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C₉-aryl "), such as indanyl or indenyl, or a ring having 10 carbon atoms (" C)₁₀-aryl "), such as tetralinyl, dihydronaphthyl or naphthyl, or is a ring having 13 carbon atoms (" C₁₃Aryl radicals ") such as the fluorenyl radical, or a ring having 14 carbon atoms (" C)₁₄Aryl) such as anthracenyl.

The term "heteroaryl" is understood as preferably meaning a monovalent monocyclic, bicyclic or tricyclic aromatic ring system which has 5,6, 7,8, 9, 10, 11, 12, 13 or 14 ring atoms ("5-to 14-membered heteroaryl"), in particular 5 or 6 or 9 or 10 carbon atoms, and which comprises at least one heteroatom which may be identical or different (the heteroatom being, for example, oxygen, nitrogen or sulfur), and which, in addition, in each case may be benzo-fused. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl (thia-4H-pyrazoyl), and the like, and benzo derivatives thereof, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, and the like; or azocinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, xanthenyl or oxepinyl (oxapinyl) and the like.

In general and unless otherwise indicated, the heteroaryl or heteroarylene includes all possible isomeric forms thereof, e.g., positional isomers thereof. Thus, for some illustrative, non-limiting examples, the term pyridyl or pyridinylene includes pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, pyridinylene-3-yl, pyridin-4-yl and pyridinylene-4-yl; alternatively, 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-C₆-alkyl group "," C₁-C₆-haloalkyl "," C₁-C₆-alkoxy "or" C₁-C₆The term "C" as used in the context of the definition of-haloalkoxy₁-C₆"is understood to mean an alkyl group having a limited number of carbon atoms from 1 to 6, i.e. 1,2, 3,4, 5 or 6 carbon atoms. It is also understood that the term "C" refers to₁-C₆"is to be understood as meaning any subrange comprised therein, such as 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-C₆-haloalkyl "or" C₁-C₆In the case of a haloalkoxy group, more particularly C₁-C₂。

Similarly, as used herein, throughout this document, for example at "C₂-C₆-alkenyl "and" C₂-C₆The term "C" as used in the context of the definition of-alkynyl₂-C₆"is understood to mean alkenyl or alkynyl groups having a limited number of carbon atoms of 2 to 6, i.e. 2,3,4, 5 or 6 carbon atoms. It is also understood that the term "C" refers to₂-C₆"is to be understood as meaning any subrange comprised therein, such as C₂-C₆、C₃-C₅、C₃-C₄、C₂-C₃、C₂-C₄、C₂-C₅(ii) a In particular C₂-C₃。

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 a limited number of carbon atoms of from 3 to 6, i.e. 3,4, 5 or 6 carbon atoms. It is also understood that the term "C" refers to₃-C₆"is to be understood as meaning any subrange comprised therein, such as 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 of the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency in the current situation 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., the substituent replaces 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 the present invention, is to be understood as meaning "one, two, three, four or five, in particular one, two, three or four, more particularly one, two or three, even more particularly one or two".

The invention also includes all suitable isotopic variations of the compounds of the invention. Definition of isotopic variations of the compounds of the present inventionIs a compound 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 present invention, for example, in which one or more members such as³H 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. Furthermore, substitution with isotopes such as deuterium may afford some therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and hence may be 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 methods or preparations 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 connects 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 or sulfoximine of the structure:

for example

Where indicates the atoms that may be bound 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, pure 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 an optically active acid or base, 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 diastereomeric 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, either as single stereoisomers or as any mixture of said isomers (e.g. R-or S-isomers or E-or Z-isomers) in any proportion. The separation of single stereoisomers, such as single enantiomers or single 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 amount 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 amount 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 as structural element of the crystal lattice of the compound a polar solvent, such as 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-) solvates or hydrates, (semi- (hemi-) solvates or hydrates, mono-, sesqui-, di-, tri-, tetra-, penta-, etc. solvates or hydrates, respectively. The present invention includes all such hydrates or solvates.

In addition, the compounds of the invention may be present in free form, for example in the form of a free base, a free acid or a zwitterion, or 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 the 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, glucoheptylic 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, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl 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.

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-carbonylmethyl (1,3-dioxolen-2-onylmethyl ester), such as 5-methyl-1, 3-dioxole-2-carbonylmethyl 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), [ α ] 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 groups (to form alkyl carbonates), dialkylcarbamoyl and N- (dialkylaminoethyl) -N-alkylcarbamoyl groups (to form carbamates), dialkylaminoacetyl and carboxyacetyl 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 said first aspect, the present invention covers a compound of general formula (I) as defined above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, optionally substituted once independently of each other by R substituents oraryl-C optionally substituted one or more times by R substituents₁-C₆-alkoxy-, heteroaryl optionally substituted one or more times by R substituents-, -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’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-an alkyl-S-group;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

r4 represents a substituent selected from:

hydrogen atom, halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl group, C₃-C₁₀-cycloalkyl-, aryl-, heteroaryl-groups;

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0,1, 2 or 3.

According to a variant of the second embodiment of said first aspect, the present invention covers a compound of general formula (I) above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, aryl optionally substituted one or more times by R substituents, independently of each other-C₁-C₆-alkoxy-, heteroaryl optionally substituted one or more times by R substituents-, -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’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-an alkyl-S-group;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

r4 represents a substituent selected from:

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0,1, 2 or 3.

According to a third embodiment of said first aspect, the present invention covers a compound of general formula (I) as defined above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

r1 represents optionally substituted by one or more substituents independently selected fromLinear chain of generations C₂-C₆Alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-a cycloalkyl group:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, aryl optionally substituted one or more times by R substituents independently of each other-C₁-C₆-alkoxy-, heteroaryl optionally substituted one or more times by R substituents-, -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’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -OC (═ O) NH₂、-OC(＝O)NHR’、-OC(＝O)N(R’)R”、-SH、C₁-C₆-an alkyl-S-group;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, -NHR', -OH, C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-an alkoxy-group;

r4 represents a substituent selected from:

hydrogen atom, halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₁₀-cycloalkyl-, aryl-, heteroaryl-groups;

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0 or 1.

According to a variant of the third embodiment of said first aspect, the present invention covers a compound of general formula (I) above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, -OH, C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-a group;

r4 represents a substituent selected from:

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0 or 1.

According to a fourth embodiment of said first aspect, the present invention covers a compound of general formula (I) as defined above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

halogen atom, -CN, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, C₃-C₆-cycloalkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, aryl optionally substituted once or twice independently of each other by R substituents, aryl optionally substituted once or more times independently of each other by R substituents-C₁-C₆-alkoxy-, heteroaryl optionally substituted once or twice independently of each other by R substituents-, -C (═ O) NH₂、-NH₂、-NHR’、-N(R’)R”、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-,-NHR’、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-an alkoxy-group;

r4 represents a substituent selected from:

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0 or 1.

According to a variant of the fourth embodiment of said first aspect, the present invention covers a compound of general formula (I) above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

halogen atom, -CN, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, C₃-C₆-cycloalkyl-, aryl optionally substituted once or twice independently of each other by R substituents, aryl optionally substituted once or more independently of each other by R substituents-C₁-C₆-alkoxy-, heteroaryl optionally substituted once or twice independently of each other by R substituents-, -C (═ O) NH₂、-NH₂、-NHR’、-N(R’)R”、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, -OH, C₁-C₆-alkoxy-, C₁-C₆-a haloalkoxy-group;

r4 represents a substituent selected from:

r represents a substituent selected from the group consisting of:

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-;

n represents an integer of 0 or 1.

According to a fifth embodiment of said first aspect, the present invention covers a compound of general formula (I) as defined above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

3-to 6-membered heterocycloalkyl linked as a spiro ring; aryl-optionally substituted once or twice independently of each other by R substituents; aryl-C optionally substituted one or more times independently of each other by R substituents₁-C₆-alkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

C₁-C₆-alkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, -NHR', -OH groups;

r4 represents a hydrogen atom;

n represents an integer of 0 or 1.

According to a variant of the fifth embodiment of said first aspect, the present invention covers a compound of general formula (I) as defined above, or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same, wherein:

represents a group selected from:

r1 represents a straight-chain C optionally substituted by one or more substituents selected independently of one another from the group consisting of₂-C₆-an alkyl-group:

aryl-optionally substituted once or twice independently of each other by R substituents; aryl-C optionally substituted one or more times independently of each other by R substituents₁-C₆-alkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

C₁-C₆-an alkoxy-group;

r4 represents a hydrogen atom;

n represents an integer of 0 or 1.

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

represents a group selected from:

wherein indicates the point of attachment of the group to the rest of the molecule.

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, aryl optionally substituted one or more times by R substituents independently of each other-C₁-C₆-alkoxy-, heteroaryl optionally substituted one or more times by R substituents-, -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’、-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-groups.

r1 tableRepresents a linear C optionally substituted by one or more substituents independently selected from₂-C₆Alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-a cycloalkyl group:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, aryl optionally substituted one or more times by R substituents, independently of each other-C₁-C₆-alkoxy-; heteroaryl which is optionally substituted one or more times by R substituents-, -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’、-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-groups.

r2 represents a hydrogen atom.

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, -OC (═ O) R', -SH, C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂N (R ') R ", -S (═ O) (═ NR') R".

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-OH、C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, -OC (═ O) R', -SH, C₁-C₆-alkyl-S-, -S (═ O) R', -S (═ O)₂R’、-S(＝O)₂NH₂、-S(＝O)₂NHR’、-S(＝O)₂N(R’)R”、-S(＝O) (═ NR') R "groups.

r4 represents a substituent selected from:

hydrogen atom, halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 10-membered heterocycloalkyl-, aryl optionally substituted one or more times by R substituents, heteroaryl optionally substituted one or more times by R substituents, C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-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 ", -S (═ O) (═ NR') R".

r represents a substituent selected from the group consisting of:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-alkenyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3-to 10-membered heterocycloalkyl-, aryl-, heteroaryl-, -C (═ O) R', -C (═ O) NH₂、-C(＝O)N(H)R’、-C(＝O)N(R’)R”、-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’、-NO₂、-N(H)S(＝O)R’、-N(R’)S(＝O)R’、-N(H)S(＝O)₂R’、-N(R’)S(＝O)₂R’、-N＝S(＝O)(R’)R”、-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 ", -S (═ O) (═ NR') R".

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

C₁-C₆-alkyl-, C₁-C₆-haloalkyl-.

n represents an integer of 0,1, 2 or 3.

R4 represents a substituent selected from:

hydrogen atom, halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl group, C₃-C₁₀-cycloalkyl-, aryl-, heteroaryl-groups.

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, -OH, C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, -NHR' -groups;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, -OH, C₁-C₆-alkoxy-, C₁-C₆-haloalkoxy-groups.

n represents an integer of 0 or 1.

halogen atom, -CN, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, C₃-C₆-cycloalkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, optionally substituted once independently of each other by R substituentsOr twice aryl-, aryl-C optionally substituted one or more times independently of each other by R substituents₁-C₆-alkoxy-, heteroaryl optionally substituted once or twice independently of each other by R substituents-, -C (═ O) NH₂、-NH₂、-NHR’、-N(R’)R”、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-.

halogen atom, -CN, C₁-C₃-alkyl-, C₁-C₃-haloalkyl-, C₃-C₆-cycloalkyl-, aryl optionally substituted once or twice independently of each other by R substituents, aryl optionally substituted once or more independently of each other by R substituents-C₁-C₆-alkoxy-, heteroaryl optionally substituted once or twice independently of each other by R substituents-, -C (═ O) NH₂、-NH₂、-NHR’、-N(R’)R”、-OH、C₁-C₃-alkoxy-, C₁-C₃-haloalkoxy-.

represents a group selected from:

3-to 6-membered heterocycloalkyl connected as a spiro ring, aryl-optionally substituted once or twice independently of each other by R substituents; optionally substituted one or more times by R substituents independently of each otheraryl-C₁-C₆-alkoxy-.

aryl optionally substituted once or twice independently of each other by R substituents, aryl optionally substituted once or more independently of each other by R substituents-C₁-C₆-alkoxy-.

r3 represents a substituent selected from:

C₁-C₆-alkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, OH-, -NHR' -groups;

r3 represents a substituent selected from:

C₃-C₆-a cycloalkoxy-group.

r3 represents a substituent selected from:

C₃-C₆-cycloalkoxy-C₁-C₃-alkoxy-groups.

r3 represents a substituent selected from:

OH-group.

r3 represents a substituent selected from:

-NHR' -group.

r3 represents a substituent selected from:

C₁-C₆-alkoxy-groups.

r4 represents a hydrogen atom.

n represents an integer of 0.

n represents an integer of 1.

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

It will be appreciated that the present invention relates to any subcombination within any embodiment or aspect of the invention of a compound of formula (I) as described hereinbefore.

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

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

According to other aspects, the present invention encompasses intermediate compounds useful for the preparation of the compounds of the present invention of general formula (I), in particular for the processes described herein. In particular, the invention encompasses compounds of general formula (V):

wherein A, R2, R3, R4 and n are as defined above for the compound of general formula (I), and X represents a leaving group, for example a halogen atom such as a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group such as a trifluoromethylsulfonate group or a nonafluorobutylsulfonate group.

According to a further aspect, the present invention encompasses the use of an intermediate compound of general formula (V), for example for the preparation of a compound of general formula (I) as defined above:

wherein A, R2, R3, R4 and n are as defined above for the compound of general formula (I), and X represents a leaving group, for example a halogen atom such as a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group such as a trifluoromethylsulfonate group.

Experimental part

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

Abbreviations	Means of
		DMSO	Dimethyl sulfoxide
THF	Tetrahydrofuran (THF)
		NMR	Nuclear magnetic resonance
DMF	N, N-dimethylformamide
		TFA	Trifluoroacetic acid
MS	Mass spectrometry
		R_t	Retention time
HPLC,LC	High performance liquid chromatography
		h	Hour(s)
min	Minute (min)
		PdCl₂(PPh₃)₂	Bis (triphenylphosphine) palladium dichloride (II)

Synthesis of compounds (review):

the compounds of the invention can be prepared as described in the following sections. Scheme 1 and the procedures described below illustrate a general synthetic route for the compounds of general formula (I) of the present invention and are not limiting. It will be apparent to those skilled in the art that the order of transformations illustrated in scheme 1 can be varied in a variety of ways. Thus, the order of transformations illustrated in scheme 1 is not limiting. In addition, substituent R can be achieved before and/or after the exemplified transformation¹、R²、R³、R⁴And tautomerization of any one of a. These modifications may be, for example, the introduction of protecting groups, cleavage of protecting groups, exchange, reduction or oxidation of functional groups, halogenation, metallation, substitution or other reactions known to those skilled in the art. These transformations include those that introduce functionality that allows for further interconversion of substituents. Suitable protecting Groups and their introduction and cleavage are well known to those skilled in the art (see, e.g., t.w.greene and p.g.m.wuts in Protective Groups in Organic Synthesis, 3 rd edition, Wiley 1999). Specific examples are described in subsequent paragraphs. Furthermore, it is possible that two or more successive steps may be carried out without work-up between the steps, for example a "one-pot" reaction, as is well known to the person skilled in the art.

Route 1

Wherein A, R1, R2, R3, R4 and n are as defined above, and X and Y represent a leaving group, for example, a halogen atom such as a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group such as a trifluoromethylsulfonate group or a nonafluorobutylsulfonate group.

In a first step, a compound of formula a (i.e. dichloropyridazine with a suitable X substituent) may be reacted with ammonia at elevated temperature and pressure to give a compound of general formula B [ similarly to WO200733080 (which is incorporated herein by reference in its entirety) ].

In a second step, the compound of formula B is reacted, for example, with chloroacetaldehyde diacetal (chloroacetaldehyde diacetal) or bromoacetaldehyde diacetal (bromoacetaldehyde diacetal) to give a bicyclic system C [ similarly to DE102006029447 (which is incorporated herein by reference in its entirety) ].

Activation of the 3-position of the bicyclic system to give compounds of formula D can be accomplished, for example, by bromination or iodination of compounds of formula C using N-bromo-succinimide or N-iodo-succinimide, respectively.

In a fourth step, the group A- [ R3 ] can be introduced using a suitably catalysed cross-linking reaction (for example using boric acid or tin hydride)]_nThereby obtaining the compound of the general formula E.

The compounds of formula E serve as important intermediates for the introduction of a plurality of side chains containing an alcohol function, thus obtaining imidazopyridazinyl ethers 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 desirable 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 interchanged as shown in route 2.

Route 2:

according to one embodiment, the present invention also relates to a process for the preparation of a compound of formula (I) as defined above, said process comprising the following steps: reacting an intermediate compound of formula (V):

wherein A, R2, R3, R4 and n are as defined above for the compound of general formula (I), and X represents a leaving group, for example a halogen atom such as a chlorine, bromine or iodine atom, or a perfluoroalkylsulfonate group such as a trifluoromethylsulfonate group or a nonafluorobutylsulfonate group,

with a compound of the general formula (III):

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

thereby obtaining a compound of formula (I):

wherein A, R1, R2, R3 and R4 are as defined above.

General part

The chemical Name was generated using ACD/Name Batch Version 12.01.

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 × 2.1 mm; 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 ℃; sample introduction: 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 uplmcs SQD 3001; column: acquity UPLC BEH C181.7 μm,50 × 2.1 mm; eluent A: water +0.1 vol% formic acid (95%), eluent B: acetonitrile, 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 ℃; sample introduction: 2 mu l of the solution; DAD scan: 210-400 nm; ELSD.

The method 3 comprises the following steps:

the instrument comprises the following steps: waters Acquity UPLCMS SQD; column: acquity UPLC BEH C181.7 μm,50 × 2.1mm; eluent A: water +0.05 vol% formic acid (95%), eluent B: acetonitrile +0.05 vol% formic acid (95%); gradient: 1-99% B at 0-1.6min, 99% B at 1.6-2.0 min; flow rate: 0.8 mL/min; temperature: 60 ℃; sample introduction amount: 2 mu L of the solution; DAD scan: PDA 210-; ELSD.

The method 4 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: 1-99% B at 0-1.6min, 99% B at 1.6-2.0 min; flow rate: 0.8 mL/min; temperature: 60 ℃; sample introduction amount: 2 mu L of the solution; DAD scan: 210-400 nm; ELSD.

The method 5 comprises the following steps:

the instrument comprises the following steps: waters Acquity uplmcs SQD 3001; column: acquity UPLC BEH C181.7 μm,50 × 2.1mm; eluent A: water +0.2 vol% ammonia (32%), eluent B: acetonitrile, gradient: 1-99% B at 0-1.6min, 99% B at 1.6-2.0 min; flow rate: 0.8 mL/min; temperature: 60 ℃; sample introduction amount: 2 mu L of the solution; DAD scan: 210-400 nm; ELSD.

Intermediates

Intermediate 1

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

For example, 3-bromo-6-chloroimidazo [1,2-b ] pyridazine is synthesized as described in WO 2007/147646 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 with 4.7mL (40mmol) of chloroacetaldehyde (55% in water) in 15mL of n-butanol at 120 ℃ for a period of 5 days. 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, 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 in the form of an amorphous white solid.

¹H-NMR (chloroform-d) [ ppm ]]＝7.06(d,1H)；7.79(d,1H)；7.92(d,1H)；7.96(d,1H)。

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

478mg (3.11mmol) of 6-chloroimidazo [1,2-b ] pyridazine is added to 10mL of chloroform under argon, and 664mg (3.73mmol) of N-bromosuccinimide is added while cooling on 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, saturated sodium bicarbonate solution was added, and the phases were separated. 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 for subsequent reactions without further chromatographic purification.

¹H-NMR (chloroform-d) [ ppm ]]＝7.12(d,1H)；7.79(s,1H)；7.90(d,1H)ppm。

Intermediate 2

6-chloro-3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazine

A mixture of 2.0g (16.8mmol) furo [3,2-b ] -pyridine in dry THF (100mL) was cooled to-78 ℃. 10.1mL (25.2mmol) of a 1.6M solution of n-butyllithium in hexane were added, and the resulting mixture was stirred at-78 ℃ for 1 h. 6.8mL (25.2mmol) of tributyltin chloride was added at-78 ℃. The cooling bath was removed and the reaction was stirred at room temperature overnight.

Methanol was carefully added and the solvent was evaporated. The resulting residue was purified by flash chromatography to give 7.4g of the corresponding crude 2-stannyl benzofuran product, which was used without further purification.

In an inert atmosphere, 3.0g (12.9mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine, 6.85g (16.8mmol) of crude 2-stannyl-furo [3,2-b ] pyridine, 246mg (1.29mmol) of cuprous iodide and 453mg (0.645mmol) of bis (triphenylphosphine) palladium (II) chloride are stirred in 100mL of THF at 85 ℃ in a closed pressure tube overnight. The solvent was evaporated, the resulting solid was dissolved in dichloromethane/methanol and filtered off. The solid was washed with methanol and hexane to obtain 2g of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝7.35-7.45(1H),7.57-7.64(1H),7.65-7.70(1H),8.08-8.15(1H),8.40-8.47(1H),8.47-8.52(1H),8.54-8.62(1H).

LCMS (method 3) R_t＝0.91min；MS(ESIpos)m/z＝271[M+H]⁺.

Intermediate 3

6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine

6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine was prepared in analogy to 6-chloro-3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazine starting from 314mg (1.35mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine to give 62% solid matter.

LCMS (method 2) R_t＝0.60min；MS(ESIpos)m/z＝271[M+H]⁺.

Intermediate 4

6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine

6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine was prepared in analogy to 6-chloro-3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazine starting from 2.4g (10.3mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine to give 2.64g of a solid material which was used as crude product.

LCMS (method 3) R_t＝1.24min；MS(ESIpos)m/z＝301[M+H]⁺.

Intermediate 5

6-chloro-3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine

A mixture of furo [2,3-c ] -pyridine (918mg,7.7mmol) in dry THF (45mL) was cooled to-78 ℃. A solution of n-butyllithium in hexane (4.6ml, c ═ 2.5M,11.6mmol) was added and the resulting mixture was stirred at-78 ℃ for 1 h. Tributyltin chloride (3.1mL,11.6mmol) was added at-78 ℃. The cooling bath was removed and the reaction mixture was stirred at room temperature for 2 h.

Methanol was added and the solvent was evaporated. Amino phase-silica gel chromatography afforded 1.9g of crude 2- (tributylstannyl) furo [2,3-c ] pyridine, which was used without further purification.

To a stirred solution of crude 2- (tributylstannyl) furo [2,3-c ] pyridine (1.9g) in THF (20mL) was added 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine (676mg,2.9mmol), cuprous iodide (55mg,0.29mmol), bis (triphenylphosphine) palladium (II) chloride (102mg,0.145mmol) and triphenylphosphine (38mg,0.145mmol) under an inert atmosphere. The mixture was heated to reflux for 2 h. The solvent was removed in vacuo. The residue was dissolved in a mixture of dichloromethane and methanol, filtered through an amino phase-silica gel column and the solvent removed in vacuo. The solid was obtained by chromatography on silica gel, which was triturated with a mixture of ethyl acetate and hexane to give 343mg of the title compound, which was used without further purification.

¹H-NMR (300MHz, chloroform-d) [ ppm ]]＝7.24(d,1H),7.62(d,1H),7.71(s,1H),8.07(d,1H),8.43(s,1H),8.48(d,1H),8.95(s,1H).

LCMS (method 3) R_t＝0.63min；MS(ESIpos)m/z＝271[M+H]⁺.

Intermediate 6

6-chloro-3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine

Step 1: 3.1g (78mmol) of sodium hydride (60% suspension in mineral oil) are carefully added at 0 ℃ to 4.7g (78mmol) of isopropanol in 100mL of anhydrous THF. The mixture was stirred at 0 ℃ for 15 min. 3g (19.5mmol) of 4-chlorofuro [3,2-c ] pyridine are added. The mixture was stirred at 80 ℃ for 20 h.

Water was carefully added. The volume of the resulting suspension was reduced by evaporation. Water was added. The aqueous layer was extracted continuously with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate and evaporated to give 4.6g of crude product, which was used in step 2 without further purification.

Step 2: 3.5g (19.5mmol) of the crude product of step 1 were cooled to-78 ℃ in 44mL of anhydrous THF. 11.7mL (29mmol) of a 2.5M solution of n-butyllithium in hexane were added. The mixture was stirred at-78 ℃ for 90 min. 6.8mL (29mmol) of triisopropyl borate were added at-78 ℃. The cooling bath was removed and the mixture was stirred at room temperature for 1 h.

A small amount of water was added and the solvent was evaporated to give 7.7g of crude product, which was used in step 3 without further purification.

And step 3: to 1.9g (8mmol) 3-bromo-6-chloroimidazo [1,2-b ] pyridazine in 68mL dioxane are added 1.9g (8.4mmol) of the crude product from step 2, 370mg (0.32mmol) tetrakis (triphenylphosphine) palladium (0) and 12mL of 2M aqueous sodium carbonate solution. The mixture was stirred at 100 ℃ for 18 h.

The reaction mixture was poured into a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and concentrated. The resulting solid material was dissolved in a 9:1 mixture of dichloromethane and methanol, filtered off, washed with dichloromethane and dried in vacuo to give 428mg of the title compound as a solid material. The mother liquor was concentrated and subjected to flash chromatography to give another portion of product containing material, which was again dissolved in methanol and dichloromethane to give another 316mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.38(6H),5.47(1H),7.33(1H),7.44(1H),7.53(1H),8.03(1H),8.36-8.40(2H).

LCMS (method 3) R_t＝1.43min；MS(ESIpos)m/z＝329[M+H]⁺.

Intermediate 7

6-chloro-3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine

6-chloro-3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine was prepared in analogy to 6-chloro-3- [4- (propan-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine starting from 2.8g (12.2mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine, and was dissolved in a 9:1 mixture of dichloromethane and methanol to give 1.3g of the title compound.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.03(9H),4.15(2H),7.35(1H),7.47(1H),7.53(1H),8.01(1H),8.37(1H).

LCMS (method 3) R_t＝1.59min；MS(ESIpos)m/z＝357[M+H]⁺.

Intermediate 8

6-chloro-3- [4- (cyclopropylmethoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine

6-chloro-3- [4- (cyclopropylmethoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine was prepared in analogy to 6-chloro-3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine starting from 3.5g (14.9mmol) of 3-bromo-6-chloro-imidazo [1,2-b ] pyridazine, which was dissolved in methanol to give 1.9g of the title compound.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝0.37(2H),0.51-0.64(2H),1.33(1H),4.26(2H),7.33(1H),7.43(1H),7.52(1H),8.00(1H),8.32-8.41(2H).

LCMS (method 2) R_t＝1.37min；MS(ESIpos)m/z＝341[M+H]⁺.

Intermediate 9

4-ethoxy furo [3,2-c ] pyridines

To a stirred solution of ethanol (14.7mL) in anhydrous THF (75mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 5.51g), and the mixture was stirred at 0 deg.C for 30 min. 4-Chlorofurano [3,2-c ] pyridine (5.0g) was added to the solution, and the mixture was stirred under reflux for 3 hours. Water was added, and the reaction mixture was extracted with ethyl acetate and hexane (1:1 mixture). The organic phase was washed with saturated sodium chloride solution, dried (sodium sulfate) and the solvent removed in vacuo. Chromatography on silica gel gives 5.1g of the title compound.

¹H-NMR (400MHz, chloroform-d), [ ppm ] C]＝1.48(3H),4.54(2H),6.83-6.90(1H),7.09(1H),7.57(1H),8.00(1H).

LCMS (method 2) R_t＝1.02min；MS(ESIpos)m/z＝164[M+H]⁺.

Intermediate 10

(4-ethoxyfuro [3,2-c ] pyridin-2-yl) boronic acid

To a stirred solution of 4-ethoxyfuro [3,2-c ] pyridine (5.1g) in anhydrous THF (170mL) at-78 deg.C was added a solution of n-butyllithium in hexane (18.8 mL; c ═ 2.5M). The solution was stirred at-78 ℃ for 1.5 h. Triisopropylborate (9.0g) was added at-78 ℃ and the mixture was stirred at-78 ℃ for 0.5h and warmed to room temperature over 16 h. Water was added, the reaction mixture was stirred for 15 minutes and the solvent was removed in vacuo. Water was again added, and the mixture was lyophilized to give 7.7g of the title compound as a crude product (calculated purity: 84%), which was used without further purification.

LCMS (method 2) R_t＝0.7min；MS(ESIpos)m/z＝208[M+H]⁺.

Intermediate 11

6-chloro-3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine

To a stirred mixture of 4-ethoxyfuro [3,2-c ] pyridine (6.1g) in anhydrous THF (90mL) at-78 ℃ was added a solution of n-butyllithium in hexane (22 mL; c ═ 2.5M). The solution was stirred at-78 ℃ for 1.5 h. Tributyltin chloride (19.2g) was added at-78 deg.C and the mixture was stirred at-78 deg.C for 0.5h and warmed to room temperature over 16 h. Methanol was added and the reaction mixture was stirred for 15 minutes. The mixture was filtered through a short silica gel column and the solvent was removed in vacuo. Silica gel chromatography gave 10.3g of 4-ethoxy-2- (tributylstannyl) furo [3,2-c ] pyridine as crude product, which was used without further purification.

To the stirred 3-bromo-6-chloro-imidazo [1,2-b ]]To a solution of pyridazine (3.3g) in THF (85mL) was added crude 4-ethoxy-2- (tributylstannyl) furo [3, 2-c%]Pyridine (10.3g), Pd₂(PPh₃)₂(510mg), triphenylphosphine (187mg) and cuprous iodide (271 mg). The mixture was heated to reflux for 5h, a mixture of dichloromethane and methanol (100:1) was added, the mixture was filtered through Celite and the solvent was removed in vacuo. The residue was triturated with warm ethanol to give 4.7g of the title compound as crude product, which was used without further purification.

LCMS (method 2) R_t＝1.36min；MS(ESIpos)m/z＝315[M+H]⁺.

Intermediate 12

(trans-3- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutyl) carbamic acid tert-butyl ester

To a stirred suspension of tert-butyl (trans) -3-hydroxycyclobutyl-carbamate (226mg) in anhydrous THF (7mL) and anhydrous DMF (0.7mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 48mg), and the mixture was stirred at room temperature for 30 min. 6-chloro-3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (190mg) was added to the solution, and the mixture was stirred at room temperature for 72 h. The solvent was removed in vacuo. Amino phase-silica gel chromatography followed by silica gel chromatography gave a solid which was triturated with ethyl acetate to give 110mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.36(9H),1.41(3H),2.52(4H),4.14-4.30(1H),4.48(2H),5.32(1H),7.04(1H),7.33(1H),7.37-7.47(2H),8.01(1H),8.11-8.19(2H).

LCMS (method 2) R_t＝1.37min；MS(ESIpos)m/z＝466[M+H]⁺.

Intermediate 13

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

To a stirred suspension of (2S) -2-aminopropan-1-ol (2.91g) in anhydrous THF (100mL) and anhydrous DMF (10mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 2.07g) and the mixture was stirred at 0 deg.C for 30 min. 3-bromo-6-chloroimidazo [1,2-b ] pyridazine (6.0g) was added to the solution, and the mixture was stirred at room temperature for 16 hours. Water was added and the mixture was extracted with a mixture of dichloromethane and methanol (100: 1). The organic phase was dried (sodium sulfate) and the solvent was removed in vacuo. A solid was obtained by silica gel chromatography, which was triturated with a mixture of toluene and cyclohexane to give 4.9g of the title compound.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.05(3H),1.63(2H),3.10-3.23(1H),4.06(2H),6.92(1H),7.69(1H),8.01(1H).

LCMS (method 5) R_t＝0.81min；MS(ESIpos)m/z＝271；273[M+H]⁺.

Intermediate 14

N-ethylfuro [3,2-c ] pyridin-4-amine

A stirred suspension of 4-chlorofuro [3,2-c ] pyridine (1.5g), ethylamine hydrochloride (2.39g) and Hunig's base (5.0mL) in 2-propanol (7.5mL) was heated to 130 ℃ in a microwave oven for 20H. Half-saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried (sodium sulfate) and the solvent removed in vacuo. Chromatography on silica gel gave 793mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.15(3H),3.40(2H),6.73(1H),6.87(1H),7.03(1H),7.75(1H),7.78(1H).

LCMS (method 5) R_t＝0.86min；MS(ESIpos)m/z＝163[M+H]⁺.

Intermediate 15

Ethyl (furo [3,2-c ] pyridin-4-yl) carbamic acid tert-butyl ester

To a stirred solution of N-ethylfuro [3,2-c ] pyridin-4-amine (940mg) and Hunig's base (3.0mL) in THF (50mL) was added di-tert-butyl dicarbonate (1.52g), and the mixture was stirred at 65 ℃ for 24H. Half-saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried (sodium sulfate) and the solvent removed in vacuo. Chromatography on silica gel gave 1.38g of the title compound.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.09(3H),1.35(9H),3.80(2H),6.74(1H),7.52(1H),8.04(1H),8.25(1H).

LCMS (method 5) R_t＝1.20min；MS(ESIpos)m/z＝263[M+H]⁺.

Intermediate 16

{4- [ (tert-Butoxycarbonyl) (ethyl) amino ] furo [3,2-c ] pyridin-2-yl } boronic acid

To a stirred solution of tert-butyl ethyl (furo [3,2-c ] pyridin-4-yl) carbamate (1.86g) in anhydrous THF (20mL) at-78 deg.C was added a solution of n-butyllithium in hexane (3.8 mL; c 2.5M). The solution was stirred at-78 ℃ for 1.5 h. Triisopropyl borate (1.92g) was added at-78 ℃ and the mixture was stirred at-78 ℃ for 0.5h and allowed to warm to room temperature over 16 h. Water was added, the reaction mixture was stirred for 15 minutes, and the solvent was removed in vacuo. Water was again added and the mixture was lyophilized to give 1.98g of the title compound as a crude product, which was used without purification.

LCMS (method 5) R_t＝0.46min；MS(ESIpos)m/z＝307[M+H]⁺.

Intermediate 17

4- (Cyclobutoxy) furo [3,2-c ] pyridine

4.98g (69mmol) of cyclobutanol are added in an ice bath to 2.7g (69mmol) of sodium hydride (60% dispersion in mineral oil) in 160mL of anhydrous THF. The mixture was stirred at 0 ℃ for 15 min. 4g (26mmol) of 4-chlorofuro [2,3-c ] pyridine are added and the mixture is stirred at 80 ℃ for 24 h.

5mL of water was carefully added, and the mixture was concentrated under reduced pressure. The residue was dissolved in 200mL of water, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and evaporated.

The crude product was purified by flash chromatography to give 3.75g of the title compound as a solid.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.59-1.74(1H),1.81(1H),2.06-2.18(2H),2.39-2.48(2H),5.32(1H),6.97(1H),7.26-7.32(1H),7.97(1H),8.01(1H).

LCMS (method 2) R_t＝1.26min；MS(ESIpos)m/z＝190[M+H]+.

Intermediate 18

[4- (Cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] boronic acid

To a stirred solution of 3.7g (19.7mmol) 4-cyclobutoxyfuro [3,2-c ] pyridine in 202mL dry THF at-78 deg.C was added 11.7mL (29mmol) of a 2.5M solution of n-butyllithium in hexane. The solution was stirred at-78 ℃ for 1.5 h. 6.8mL (29mmol) of triisopropyl borate were added at-78 deg.C and the mixture was stirred at room temperature for 2 h. Water was added and the solvent was removed in vacuo to give 7.2g of the title compound as a crude product, which was used without further purification.

LCMS (method 2) R_t＝0.88min；MS(ESIpos)m/z＝234[M+H]⁺.

Intermediate 19

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

(2R) -1- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-2-amine was prepared in analogy to its enantiomer (2S) -1- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-2-amine.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.05(3H),3.17(1H),4.06(2H),6.92(1H),7.69(1H),8.01(1H).

LCMS (method 4) R_t＝0.55min；MS(ESIpos)m/z＝271；273[M+H]⁺.

Examples

Example 1

(1S) -2- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine

81mg (0.59mmol) of (S) -2-phenylglycinol ((S) -2-phenylglycinol) are added at 0 ℃ to 23mg (0.59mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 80mg (0.3mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into half-saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 29mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝4.42-4.49(1H),4.55-4.70(2H),7.04(1H),7.27-7.34(1H),7.38(2H),7.54(2H),7.67(1H),7.74(1H),8.16-8.23(3H),8.51(1H),8.98(1H).

LC-MS (method 3) R_t＝0.53min；MS(ESIpos)m/z＝372[M+H]⁺.

Example 2

Trans-3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] -oxy } cyclobutylamine

91mg (0.74mmol) of trans-3-aminocyclobutanol hydrochloride are added at 0 ℃ to 44mg (1.11mmol sodium hydride (60% in mineral oil) in 4mL dry THF, stirred on an ice bath for 15min, then 150mg (0.37mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added, the ice bath is removed, and the mixture is stirred at 40 ℃ for 72h, the mixture is cooled again to 0-5 ℃, an additional 68mg (0.56mmol) of trans-3-aminocyclobutanol hydrochloride and 29mg (0.74mmol) sodium hydride (60% in mineral oil) are added, stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 61mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝2.51-2.58(4H),3.39-3.48(2H),3.79-3.85(1H),4.06(3H),5.39-5.49(1H),7.05(1H),7.35-7.42(1H),7.48(1H),8.07(1H),8.13-8.22(2H).

LC-MS (method 3) R_t＝0.69min；MS(ESIpos)m/z＝352[M+H]⁺.

Example 3

(2R) -1- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] -oxy } propan-2-amine

39mg (0.52mmol) of (R) -2-aminopropan-1-ol are added to 21mg (0.52mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring was carried out for 15min on an ice bath, then 105mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 45mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.19(3H),3.37-3.49(1H),4.00(3H),4.19-4.39(2H),7.02(1H),7.35(1H),7.42(1H),8.03(1H),8.10-8.19(2H).

LC-MS (method 3) R_t＝0.74min；MS(ESIpos)m/z＝340[M+H]⁺.

Example 4

(1S) -2- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine

71mg (0.52mmol) of (S) -2-phenylglycinol are added at 0 ℃ to 21mg (0.52mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF. Stirring was carried out for 15min on an ice bath, then 105mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 19 h.

The reaction mixture was poured into half-saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 4mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝4.02(3H),4.35-4.52(2H),4.55-4.67(1H),6.98-7.09(1H),7.38(5H),7.48(1H),7.52-7.59(2H),8.02-8.09(1H),8.12-8.22(2H).

LC-MS (method 3) R_t＝0.86min；MS(ESIpos)m/z＝402[M+H]⁺.

Example 5

(2S) -1- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine

39mg (0.52mmol) of (S) -2-aminopropan-1-ol are added to 21mg (0.52mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring was carried out for 15min on an ice bath, then 105mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 46mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.19(3H),3.39-3.45(1H),4.00(3H),4.21-4.38(2H),7.02(1H),7.35(1H),7.42(1H),8.03(1H),8.11-8.18(2H).

LC-MS (method 3) R_t＝0.74min；MS(ESIpos)m/z＝340[M+H]⁺.

Example 6

(2R) -2- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] -oxy } propan-1-amine

39mg (0.52mmol) of (R) -1-aminopropan-2-ol are added to 21mg (0.52mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring was carried out for 15min on an ice bath, then 105mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 48mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.43(3H),2.93(2H),4.01(3H),5.06-5.18(1H),6.98(1H),7.35(1H),7.41(1H),8.03(1H),8.11-8.17(2H).

LC-MS (method 3) R_t＝0.76min；MS(ESIpos)m/z＝340[M+H]⁺.

Example 7

(1R) -2- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine

82mg (0.6mmol) of (R) -2-phenylglycinol are added at 0 ℃ to 24mg (0.6mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF. Stirring was carried out for 15min on an ice bath, then 81mg (0.3mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into half-saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 42mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝4.45(1H),4.53-4.69(2H),7.04(1H),7.26-7.32(1H),7.34-7.41(2H),7.54(2H),7.67(1H),7.74(1H),8.15-8.24(2H),8.50(1H),8.98(1H).

LC-MS (method 3) R_t＝0.52min；MS(ESIpos)m/z＝372[M+H]⁺.

Example 8

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

33mg (0.44mmol) of (R) -1-aminopropan-2-ol are added to 14mg (0.59mmol) of sodium hydride (60% in mineral oil) in 5mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 80mg (0.3mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 16 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The whole mixture was concentrated and purified by HPLC, flash chromatography and preparative thin layer chromatography successively to give 8mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.43(3H),2.85-2.92(2H),5.13-5.26(1H),6.97-7.04(1H),7.59-7.65(1H),7.68-7.74(1H),8.11-8.21(2H),8.43-8.51(1H),8.98-9.04(1H).

LC-MS (method 3) R_t＝0.47min；MS(ESIpos)m/z＝310[M+H]⁺.

Example 9

(1R) -2- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine

71mg (0.52mmol) of (R) -2-phenylglycinol are added at 0 ℃ to 21mg (0.52mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF. Stirring was carried out for 15min on an ice bath, then 105mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into half-saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 69mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝3.98(3H),4.42(2H),4.53-4.64(1H),7.00(1H),7.25-7.40(4H),7.43(1H),7.52(2H),8.02(1H),8.10-8.17(2H).

LC-MS (method 3) R_t＝0.88min；MS(ESIpos)m/z＝402[M+H]⁺.

Example 10

(2R,3R) -3- (benzyloxy) -1- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } butan-2-amine

84mg (0.43mmol) of (2R,3R) -2-amino-3- (benzyloxy) butan-1-ol are added at 0 ℃ to 17mg (0.43mmol) of sodium hydride (60% in mineral oil) in 3mL of anhydrous THF and 1mL of DMF. Stirring is carried out in an ice bath for 15min, then 100mg (0.22mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 17 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 20mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.24(3H),1.73-2.01(1H),3.06-3.20(1H),3.64-3.74(1H),3.93(3H),4.24-4.37(1H),4.39-4.53(2H),4.61(1H),6.98(1H),7.06-7.21(3H),7.29(2H),7.36(1H),7.49(1H),8.03(1H),8.10-8.18(2H).

LC-MS (method 3) R_t＝0.90min；MS(ESIpos)m/z＝460[M+H]⁺.

Example 11

(2R) -1- (benzyloxy) -3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] -pyridazin-6-yl ] oxy } propan-2-amine

196mg (1.1mmol) of (R) -2-amino-3-benzyloxypropan-1-ol was added to 43mg (1.1mmol) of sodium hydride (60% in mineral oil) in 8mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 250mg (0.54mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 16 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 38mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝3.37(1H),3.51(2H),3.95(3H),4.35(1H),4.47(1H),4.53(2H),7.01(1H),7.15-7.33(5H),7.36(1H),7.48(1H),8.03(1H),8.15(2H).

LC-MS (method 3) R_t＝0.87min；MS(ESIpos)m/z＝446[M+H]⁺.

Example 12

(2S) -1- { [3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine

To a stirred suspension of (2S) -2-aminopropan-1-ol (27mg, 354. mu. mol) in anhydrous THF (3.5mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 23mg), and the mixture was stirred at 0 deg.C for 30 min. 6-chloro-3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (80mg, 177. mu. mol) was added to the solution, and the mixture was stirred at room temperature for 2 h. A half-saturated aqueous sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic phase was dried (sodium sulfate) and the solvent was removed in vacuo. Amino phase-silica gel chromatography afforded a solid, which was triturated with a mixture of ethanol and hexane to give 35mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆):[ppm]＝1.16(d,3H),1.72(br.s.,2H),3.24-3.32(m,1H),4.28(d,2H),7.12(d,1H),7.65(d,1H),7.78(dd,1H),8.22(d,1H),8.28(s,1H),8.43(d,1H),8.99(s,1H).

LCMS (method 3) R_t＝0.74min；MS(ESIpos)m/z＝310[M+H]⁺.

Example 13

Trans-3- { [3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine formate salt

To a stirred suspension of trans-3-aminocyclobutanol hydrochloride (57.5mg, 465. mu. mol) in anhydrous THF (3.0mL) and anhydrous DMF (1.5mL) at 0 ℃ was added sodium hydride (60% w/w in oil; 27mg) and the mixture was stirred at 0 ℃ for 30 minutes. 6-chloro-3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (70mg, 155. mu. mol) was added to the solution, and the mixture was stirred at room temperature for 16 h. Half-saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The solvent was removed in vacuo. Amino phase-silica gel chromatography afforded a solid, which was triturated with dichloromethane. Preparative reverse phase HPLC gave 21mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆Detection signal of formate) [ ppm ]]＝2.52-2.69(m,4H),3.79(br.s.,1H),5.49-5.67(m,1H),7.09(d,1H),7.67(s,1H),7.79(d,1H),8.22(d,1H),8.27(s,1H),8.37(br.s.,1H),8.44(d,1H),8.98(s,1H).

LCMS (method 3) R_t＝0.47min；MS(ESIpos)m/z＝322[M+H]⁺.

Example 14

Trans-3- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine formate salt

To a stirred suspension of trans-3-aminocyclobutanol hydrochloride (110mg) in anhydrous THF (6mL) and anhydrous DMF (3mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 52mg) and the mixture was stirred at 0 deg.C for 30 min. 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (80mg) and potassium carbonate (204mg) were added to the solution, and the mixture was stirred at room temperature for 72 h. Half-saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. Amino phase-silica gel chromatography followed by preparative reverse phase HPLC gave a solid which was triturated with dichloromethane to give 40mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆Detection signal for formate), [ ppm]＝2.50-2.64(4H),3.76(1H),5.59(1H),7.04(1H),7.66-7.74(2H),8.13-8.21(2H),8.39(1H),8.46(1H),9.03(1H).

LC-MS (method 2) R_t＝0.47min；MS(ESIpos)m/z＝322[M+H]⁺.

Example 15

(2R) -2-amino-3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-ol

To a solution of 60mg (0.14mmol) of (2R) -1- (benzyloxy) -3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine in 5mL of methanol were added 43mg of palladium on charcoal (containing 10% palladium) and 0.67. mu.L of 4M hydrochloric acid/dioxane. The flask was purged with hydrogen and equipped with a hydrogen balloon. The mixture was stirred vigorously for 1 day.

The catalyst was filtered off and washed with methanol. The filtrate was evaporated and the resulting crude product was purified by HPLC to give 5mg of the title compound as a solid material.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝3.15-3.24(2H),3.43-3.55(2H),4.02(3H),4.28-4.37(1H),4.39-4.49(1H),6.99-7.07(1H),7.33-7.39(1H),7.48(1H),8.02-8.07(1H),8.13-8.19(2H).

LCMS (method 3) R_t＝0.65min；MS(ESIpos)m/z＝356[M+H]⁺.

Example 16

3- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-methylpropan-1-amine

96mg (1.1mmol) of 3-amino-2-methylpropan-1-ol are added at 0 ℃ to 43mg (1.1mmol) of sodium hydride (60% in mineral oil) in 8mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 250mg (0.54mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 122mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.07(3H),4.01(3H),4.41(2H),7.04(1H),7.36(1H),7.50(1H),8.04(1H),8.12-8.20(2H).

LC-MS (method 3) R_t＝0.70min；MS(ESIpos)m/z＝354[M+H]⁺.

Example 17

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

41mg (0.56mmol) of (R) -2-aminopropan-1-ol are added to 22mg (0.56mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 80mg (0.29mmol) of 6-chloro-3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into a saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 53mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.19(3H),3.39(1H),4.29-4.39(2H),7.08(1H),7.34(1H),7.66(1H),8.05(1H),8.20(1H),8.23(1H),8.51(1H).

LC-MS (method 3) R_t＝0.57min；MS(ESIpos)m/z＝310[M+H]⁺.

Example 18

(1S,3R) -3- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclopentylamine

36mg (0.26mmol) of (1R,3S) -3-aminocyclopentanol hydrochloride are added to 16mg (0.39mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring was carried out for 15min on an ice bath, then 129mg (0.26mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine were added. The ice bath was removed and the mixture was stirred at 40 ℃ for 15 h.

0.07mL (0.53mmol) of triethylamine was added and the mixture was stirred at 40 ℃ for a further 7 h.

In a separate flask, 19mg (0.13mmol) of (1R,3S) -3-aminocyclopentanol hydrochloride are added to 8mg (0.2mmol) of sodium hydride (60% in mineral oil) in 1mL of anhydrous DMF at 0 ℃. The mixture was added to the reaction and the resulting mixture was stirred at 40 ℃ for a further 16 h.

In a separate flask, 19mg (0.13mmol) of (1R,3S) -3-aminocyclopentanol hydrochloride are added to 8mg (0.2mmol) of sodium hydride (60% in mineral oil) in 1mL of anhydrous DMF at 0 ℃. The mixture was again added to the reaction and the resulting mixture was stirred at 40 ℃ for a further 16 h.

The reaction mixture was poured into a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 54mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.65(1H),1.72-1.81(1H),1.97(1H),2.06-2.15(2H),2.52-2.61(1H),3.46(1H),4.04(3H),5.37-5.44(1H),7.02(1H),7.38(1H),7.48(1H),8.06(1H),8.14-8.19(2H).

LC-MS (method 3) R_t＝0.77min；MS(ESIpos)m/z＝366[M+H]⁺.

Example 19

(2S) -1- ({3- [4- (propan-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine

42mg (0.55mmol) of (2S) -aminopropan-1-ol are added to 22mg (0.55mmol) of sodium hydride (60% in mineral oil) in 3.6mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 90mg (0.27mmol) of 6-chloro-3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by HPLC to give 49mg of the title compound as a solid substance.

¹H-NMR(600MHz,DMSO-d₆):[ppm]＝1.20(3H),1.39(6H),3.47(1H),4.21(1H),4.41(1H),5.45(1H),7.09(1H),7.35(1H),7.51(1H),8.05(1H),8.18(1H),8.21(1H).

LC-MS (method 3) R_t＝0.88min；MS(ESIpos)m/z＝368[M+H]⁺.

Example 20

(2S) -1- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine

44mg (0.59mmol) of (2S) -2-aminopropan-1-ol are added to 24mg (0.59mmol) of sodium hydride (60% in mineral oil) in 5mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 80mg (0.27mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 16 h.

The reaction mixture was poured into a saturated ammonium chloride solution and extracted with ethyl acetate. The organic layer was concentrated and purified by HPLC to give 10mg of the title compound as a solid material.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.13(3H),4.26(2H),7.06(1H),7.64-7.74(2H),8.12-8.25(2H),8.47(1H),9.01(1H).

Example 21

Trans-4- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] -oxy } cyclohexaneamine formate salt

To a stirred suspension of trans-4-aminocyclohexanol hydrochloride (56mg) in anhydrous THF (2mL) and anhydrous DMF (2mL) at 0 ℃ was added sodium hydride (60% w/w in oil; 31mg) and the mixture was stirred at 0 ℃ for 30 minutes. 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (75mg) was added to the solution, and the mixture was stirred under reflux for 30 minutes. The solid was removed by filtration and the solvent was removed in vacuo. The residue was dissolved in DMSO and formic acid (100: 0.1). Preparative reverse phase HPLC gave a solid which was triturated with ethanol to give 60mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆Signal of formate), [ ppm]＝1.35-1.66(4H),1.99(2H),2.33(2H),2.80-2.97(1H),4.02(3H),4.83-5.01(1H),6.99(1H),7.35(1H),7.47(1H),8.03(1H),8.10-8.20(2H),8.44(4H).

LC-MS (method 3) R_t＝0.80min；MS(ESIpos)m/z＝380[M+H]⁺.

Example 22

1- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-methylpropan-2-amine

59mg (0.66mmol) of 2-amino-2-methylpropan-1-ol are added to 27mg (0.67mmol) of sodium hydride (60% in mineral oil) in 5mL of anhydrous DMF at 0-5 ℃. Stirring is carried out for 5min on an ice bath, then 100mg (0.33mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and stirred at room temperature for 3 hours.

The reaction mixture was poured into a half-saturated ammonium chloride solution. Extracted four times with ethyl acetate. The combined organic phases were washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 55mg (47%) of product.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.18(6H),4.01(3H),4.18(2H),7.05(1H),7.36(1H),7.45(1H),8.04(1H),8.11-8.19(2H).

LC-MS (method 2) R_t＝0.73min；MS(ESIpos)m/z＝353[M+H]⁺.

Example 23

3- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-phenylpropan-1-amine

203mg (1.1mmol) of 3-amino-2-phenylpropan-1-ol hydrochloride are added at 0 ℃ to 86mg (2.1mmol) of sodium hydride (60% in mineral oil) in 8mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 250mg (0.54mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by HPLC to give 83mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝3.08-3.14(1H),3.15-3.24(1H),3.37-3.50(1H),4.03(3H),4.65-4.85(2H),6.99-7.10(1H),7.25-7.46(6H),7.51-7.58(1H),8.03-8.11(1H),8.13-8.22(2H).

LC-MS (method 3) R_t＝0.80min；MS(ESIpos)m/z＝416[M+H]⁺.

Example 24

Trans-3- ({3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) cyclobutylamine

94mg (0.5mmol) of tert-butyl (trans-3-hydroxycyclobutyl) carbamate are added at 0 ℃ to 20mg (0.5mmol) of sodium hydride (60% in mineral oil) in 6mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 90mg (0.25mmol) of 6-chloro-3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 19 h.

The reaction mixture was poured into a half-saturated aqueous sodium chloride solution and extracted with dichloromethane. The organic layer was dried over sodium sulfate and concentrated.

The resulting crude material was dissolved in 10mL of dichloromethane. 5mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 10 min.

5mL of ammonia (25% in water) was carefully added. Half-saturated aqueous sodium chloride solution was added. The mixture was extracted with dichloromethane. The organic layer was dried over sodium sulfate and concentrated.

The crude material was purified by HPLC to give 30mg of the title compound as a solid material.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.08(9H),2.24-2.37(2H),2.54(2H),3.66-3.78(1H),4.15(2H),5.34-5.45(1H),7.06(1H),7.37(1H),7.55(1H),8.03(1H),8.15-8.22(2H).

LC-MS (method 4) R_t＝0.96min；MS(ESIpos)m/z＝408[M+H]⁺.

Example 25

(2S) -1- ({3- [4- (cyclopropylmethoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine

41mg (0.54mmol) of (2S) -2-aminopropan-1-ol are added to 21mg (0.54mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 120mg (0.27mmol) of 6-chloro-3- [ 4-cyclopropylmethoxy) -furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 18 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was concentrated and purified by HPLC, then by flash chromatography to give 40mg of the title compound as a solid material.

¹H-NMR(400MHz,DMSO-d₆):[ppm]＝0.38-0.44(2H),0.55-0.62(2H),1.15-1.21(3H),1.29-1.37(1H),3.39-3.45(1H),4.20(1H),4.32(2H),4.37(1H),7.08(1H),7.37(1H),7.53(1H),8.03(1H),8.18(1H),8.20(1H).

LC-MS (method 2) R_t＝0.83min；MS(ESIpos)m/z＝380[M+H]⁺.

Example 26

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

44mg (0.59mmol) of (2R) -2-aminopropan-1-ol are added to 23mg (0.59mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 80mg (0.3mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 16 h.

The reaction mixture was poured into a saturated ammonium chloride solution and extracted with ethyl acetate. The organic layer was concentrated and purified by HPLC to give 19mg of the title compound as a solid material.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.17(3H),1.65-1.83(2H),3.17-3.25(1H),4.26-4.33(2H),7.07-7.13(1H),7.68-7.77(2H),8.22(2H),8.47-8.53(1H),9.01-9.08(1H).

LC-MS (method 2) R_t＝0.44min；MS(ESIpos)m/z＝310[M+H]⁺.

Example 27

1- [3- ({ [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } methyl) oxetan-3-yl ] methanamine

31mg (0.27mmol) of [3- (aminomethyl) oxetan-3-yl ] methanol was added at 0 ℃ to 11mg (0.27mmol) of sodium hydride (60% in mineral oil) in 2mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 54mg (0.13mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by HPLC to give 15mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝2.97(2H),4.02(3H),4.39-4.50(4H),4.69(2H),7.05(1H),7.36(1H),7.55(1H),8.04(1H),8.16(2H).

LC-MS (method 3) R_t＝0.69min；MS(ESIpos)m/z＝382[M+H]⁺.

Example 28

Trans-3- { [3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine

104mg (0.6mmol) of tert-butyl (trans-3-hydroxycyclobutyl) carbamate are added at 0 ℃ to 22mg (0.56mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 80mg (0.28mmol) of 6-chloro-3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into a saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated.

The resulting crude material was dissolved in 2mL of dichloromethane. 1mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 15 min.

Carefully add 2mL ammonia (25% in water). Water was added. The mixture was extracted with a 95:5 mixture of dichloromethane and methanol. The organic layer was dried over magnesium sulfate and concentrated.

The crude material was purified by HPLC to give 48mg of the title compound as a solid material.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝2.54-2.60(4H),3.80(1H),5.54(1H),7.07(1H),7.34(1H),7.65(1H),8.05(1H),8.19(1H),8.22-8.27(1H),8.52(1H).

LC-MS (method 3) R_t＝0.58min；MS(ESIpos)m/z＝322[M+H]⁺.

Example 29

Trans-3- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine

84mg (0.68mmol) of trans-3-aminocyclobutanol hydrochloride in 2mL of a 1:1 mixture of anhydrous THF and anhydrous DMF is added at 0 ℃ to 41mg (1mmol) of sodium hydride (60% in mineral oil) in 2mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 100mg (0.34mmol) of 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 72 h.

The reaction mixture was poured into water and extracted with ethyl acetate. The combined organic and aqueous phases were concentrated separately and then combined to purify the residue by HPLC to give 22mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝2.46(4H),3.55-3.72(1H),5.33-5.55(1H),6.95-7.11(1H),7.57-7.80(2H),8.10-8.26(2H),8.41-8.58(1H),8.95-9.11(1H).

LC-MS (method 3) R_t＝0.48min；MS(ESIpos)m/z＝322[M+H]⁺.

Example 30

(2S) -1- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -3-methylbutan-2-amine

To a stirred suspension of (S) - (+) -2-amino-3-methyl-1-butanol (53mg) in anhydrous THF (5mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 34mg), and the mixture was stirred at 0 deg.C for 30 min. 6-chloro-3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (70mg) was added to the solution, and the mixture was stirred at room temperature for 72 hours. Half-saturated sodium chloride solution was added, and the mixture was extracted with ethyl acetate. The solution was dried (sodium sulfate) and the solvent was removed in vacuo. Amino phase-silica gel chromatography afforded a solid, which was triturated with a mixture of ethanol and hexane to give 48mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝0.95(6H),1.52(2H),1.72-1.89(1H),2.87-2.99(1H),4.23-4.35(1H),4.36-4.47(1H),7.06(1H),7.63(1H),7.69(1H),8.09-8.21(2H),8.46(1H),8.97(1H).

LC-MS (method 2) R_t＝0.51min；MS(ESIpos)m/z＝338[M+H]⁺.

Example 31

(1S,2S) -2- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclopentylamine

137mg (1.00mmol) of (1S,2S) -2-aminocyclopentanol hydrochloride are added to 79.8mg (2.00mmol) of sodium hydride (60% in mineral oil) in 7mL of anhydrous DMF at 0-5 ℃. After stirring for 5 minutes on an ice bath, 150mg (0.50mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and stirred at room temperature for 3 h. The reaction mixture was poured into water. It was concentrated. To the residue were added 1mL of DMF, 3mL of methanol and 0.5mL of water. Heat was applied at reflux and insoluble material was filtered from the hot solution using a Whatman filter. The filtrate was concentrated and dissolved in a mixture of 1mL DMF and 3mL methanol. The insoluble material was filtered and discarded. The filtrate was purified by HPLC to give 43mg (23%) of product.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.37-1.49(1H),1.61-1.88(3H),1.88-2.03(1H),2.25-2.38(1H),3.40-3.48(1H),4.01(3H),4.96-5.03(1H),6.98(1H),7.36(1H),7.53(1H),8.03(1H),8.10-8.18(2H).

LC-MS (method 2) R_t＝0.76min；MS(ESIpos)m/z＝365[M+H]⁺.

Example 32

(2S) -1- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine

To a stirred mixture of (2S) -1- [ (3-bromoimidazo [1,2-b ]]Pyridazin-6-yl) oxy]Method for preparing propan-2-amine (5.60g) in 1-propanol (100mL)To the solution were added 2M potassium carbonate solution (31ml), crude (4-ethoxyfuro [3,2-c ]]Pyridin-2-yl) boronic acid (84% w/w; 7.64g), triphenylphosphine (542mg) and PdCl₂(PPh₃)₂(1.45 g). The mixture was heated to reflux for 2 h. The warm mixture was filtered through Celite and the solvent removed in vacuo. Half-saturated sodium bicarbonate solution was added and the mixture was extracted with a mixture of dichloromethane and methanol. The organic phase was washed with saturated sodium chloride solution, dried (sodium sulfate) and the solvent removed in vacuo. A solid was obtained by silica gel chromatography, which was triturated with a mixture of dichloromethane and hexane to give 4.17g of the title compound.

¹H-NMR(300MHz,DMSO-d₆Detection signal), [ ppm ] ppm]＝1.13(3H),1.36(3H),3.29-3.42(1H),4.14(1H),4.29(1H),4.45(2H),7.02(1H),7.31(1H),7.42(1H),7.99(1H),8.10-8.17(2H).

LC-MS (method 5) R_t＝1.04min；MS(ESIpos)m/z＝354[M+H]⁺.

Example 33

2- { [3- (4-Methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -3-phenylpropan-1-amine

124.5mg (0.66mmol) of 1-amino-3-phenylpropan-2-ol hydrochloride are added at 0-5 ℃ to 58.5mg (1.46mmol) of sodium hydride (60% in mineral oil) in 4.5mL of anhydrous DMF. After stirring for 5 minutes on an ice bath, 100mg (0.33mmol) of 6-chloro-3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine are added. The ice bath was removed and stirred at room temperature for 3 h. The reaction mixture was poured into a half-saturated ammonium chloride solution. 25mL of ethyl acetate was added, and the layers were separated. The insoluble material in the aqueous phase was filtered off and washed with ethyl acetate. The aqueous phase was extracted three times with ethyl acetate. The combined organic phases were washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by HPLC to give 9mg (6%) of product.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝2.79-3.08(3H),3.17-3.25(1H),4.05(3H),5.31-5.42(1H),7.00(1H),7.16-7.28(3H),7.29-7.39(3H),7.51(1H),8.06(1H),8.11-8.19(2H).

LC-MS (method 4) R_t＝0.89min；MS(ESIpos)m/z＝415[M+H]+.

Example 34

(2S) -1- ({3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine

39mg (0.5mmol) of (2S) -2-aminopropan-1-ol are added to 20mg (0.5mmol) of sodium hydride (60% in mineral oil) in 4mL of anhydrous THF at 0 ℃. Stirring is carried out in an ice bath for 15min, then 90mg (0.25mmol) of 6-chloro-3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 19 h.

The reaction mixture was poured into water and extracted successively with dichloromethane and ethyl acetate. The combined organic layers were dried over sodium sulfate and concentrated. The resulting substance was dissolved in methanol to obtain 64mg of the title compound as a solid substance.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.06(9H),1.13(3H),1.69(2H)3.34-3.38(1H),4.12-4.18(3H),4.35(1H),7.09(1H),7.36(1H),7.58(1H),8.03(1H),8.18(1H),8.20(1H.)

LC-MS (method 3) R_t＝0.98min；MS(ESIpos)m/z＝396[M+H]⁺.

Example 35

2- {6- [ (trans-3-aminocyclobutyl) oxy ] imidazo [1,2-b ] pyridazin-3-yl } furo [3,2-c ] -pyridin-4-ol

To 325mg (0.93mmol) of trans-3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo- [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine in 5mL of dioxane is added 0.46mL of a 4M HCl/dioxane solution. The mixture was stirred at room temperature for 1 h.

The solvent was evaporated. The resulting material was dissolved in methanol. The resulting solid substance was subjected to HPLC purification to obtain 43mg of the title compound as a solid substance.

¹H-NMR(500MHz,DMSO-d₆):[ppm]＝2.58-2.72(3H),3.91(1H),5.42-5.50(1H),6.75(1H),7.03(1H),7.38(1H),7.44(1H),8.09(1H),8.18(1H).

LC-MS (method 3) R_t＝0.98min；MS(ESIpos)m/z＝396[M+H]⁺.

Example 36

Trans-3- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine

To a stirred suspension of tert-butyl (trans-3- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutyl) carbamate (110mg) in dichloromethane (10mL) was added TFA (0.5 mL). The mixture was stirred at room temperature for 16 h. TFA (1mL) was added again, and the mixture was stirred at room temperature for 72 h. Saturated potassium carbonate solution was added until a pH of 9 was reached. The mixture was extracted with dichloromethane and methanol (10:1 mixture). The solution was dried (sodium sulfate) and the solvent was removed in vacuo. Chromatography on silica gel gave 40mg of the title compound.

¹H-NMR(300MHz,DMSO-d₆Detection signal), [ ppm ] ppm]＝1.34-1.45(3H),2.11-2.38(4H),3.58-3.73(1H),4.46(2H),5.31-5.46(1H),7.01(1H),7.32(1H),7.50(1H),8.00(1H),8.10-8.18(2H).

LC-MS (method 2) R_t＝0.74min；MS(ESIpos)m/z＝366[M+H]⁺.

Example 37

Trans-3- ({3- [4- (propan-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) cyclobutylamine

103mg (0.55mmol) of tert-butyl (trans-3-hydroxycyclobutyl) carbamate are added at 0 ℃ to 22mg (0.55mmol) of sodium hydride (60% in mineral oil) in 5mL of anhydrous THF. Stirring is carried out in an ice bath for 15min, then 90mg (0.27mmol) of 6-chloro-3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazine are added. The ice bath was removed and the mixture was stirred at 40 ℃ for 16 h.

5mL of ammonia (25% in water) was carefully added. Half-saturated aqueous sodium chloride solution was added. The mixture was extracted with dichloromethane. The organic layer was dried over magnesium sulfate and concentrated.

The crude material was purified by flash chromatography to give 76mg of the title compound as a solid material.

¹H-NMR(500MHz,DMSO-d₆):[ppm]＝1.42(6H),2.35-2.43(2H),3.70-3.77(1H),5.46(2H),7.04(1H),7.33(1H),7.54(1H),8.04(1H),8.16(1H),8.18(1H).

LC-MS (method 3) R_t＝0.78min；MS(ESIpos)m/z＝380[M+H]⁺.

Example 38

(2R) -1- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine

To a stirred suspension of (2R) -2-aminopropan-1-ol (48mg) in dry THF (6mL) at 0 deg.C was added sodium hydride (60% w/w in oil; 42mg), and the mixture was stirred at room temperature for 30 min. 6-chloro-3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazine (100mg) was added to the solution, and the mixture was stirred at room temperature for 16 h. Ethanol was added carefully, the mixture was stirred for 5 minutes, and the solvent was removed in vacuo. Amino phase-silica gel chromatography followed by silica gel chromatography gave 45mg of the title compound.

1H-NMR(400MHz,DMSO-d6),[ppm]＝1.13(3H),1.37(3H),2.01(2H),3.31-3.41(1H),4.16(1H),4.29(1H),4.46(2H),7.03(1H),7.32(1H),7.45(1H),8.01(1H),8.11-8.18(2H).

LC-MS (method 2) R_t＝0.76min；MS(ESIpos)m/z＝354[M+H]⁺.

Example 39

[2- (6- { [ (2S) -2-aminopropyl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) furo [3,2-c ] pyridin-4-yl ] ethylcarbamic acid tert-butyl ester

To a stirred mixture of (2S) -1- [ (3-bromoimidazo [1,2-b ]]Pyridazin-6-yl) oxy]To a solution of propan-2-amine (130mg) in 1-propanol (13ml) was added 2M potassium carbonate solution (0.7ml), crude {4- [ (tert-butoxycarbonyl) (ethyl) amino group]Furo [3,2-c ]]Pyridin-2-yl } boronic acid (70% w/w; 416mg), triphenylphosphine (12.5mg) and PdCl₂(PPh₃)₂(33.5 mg). The mixture was heated to reflux for 1 h. The warm mixture was filtered through Celite and the solvent was removed in vacuo. Half-saturated sodium bicarbonate solution was added and the mixture was extracted with a mixture of dichloromethane and methanol. The organic phase was washed with saturated sodium chloride solution, dried (sodium sulfate) and the solvent removed in vacuo. The solid was obtained by silica gel chromatography, which was triturated with a mixture of dichloromethane and hexane to give 125mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆),[ppm]＝1.12(3H),1.17(3H),1.38(9H),1.97(2H),3.30-3.34(1H),3.85(2H),4.11(1H),4.27(1H),7.10(1H),7.35(1H),7.63(1H),8.17-8.25(2H),8.33(1H).

LC-MS (method 5) R_t＝1.13min；MS(ESIpos)m/z＝453[M+H]⁺.

Example 40

2- (6- { [ (2S) -2-aminopropyl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) -N-ethylfuro [3,2-c ] pyridin-4-amine

To a stirred suspension of tert-butyl [2- (6- { [ (2S) -2-aminopropyl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) furo [3,2-c ] pyridin-4-yl ] ethylcarbamate (115mg) in dichloromethane (1mL) was added TFA (0.4 mL). The mixture was stirred at room temperature for 4 h. The solvent was removed in vacuo. The residue was dissolved in dichloromethane and methanol and saturated potassium carbonate solution was added until pH9 was reached. The organic phase was separated and dried (sodium sulfate) and the solvent was removed in vacuo. Chromatography on silica gel gave a solid which was triturated with methanol to give 83mg of the title compound.

¹H-NMR(400MHz,DMSO-d₆Detection signal), [ ppm ] ppm]＝1.13(3H),1.20(3H),1.66(2H),3.42-3.53(2H),4.24-4.36(2H),6.83(1H),7.00(1H),7.10(1H),7.68(1H),7.87(1H),8.05(1H),8.12(1H).

LC-MS (method 5) R_t＝0.93min；MS(ESIpos)m/z＝353[M+H]⁺.

EXAMPLE 41

(2S) -1- ({3- [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine

To a stirred solution of 100mg (0.37mmol) of (2S) -1- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-2-amine in 6mL 1-propanol were added 550. mu.L (1.1mmol) of a 2M solution of potassium carbonate, 344mg (0.74mmol) of the crude [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] boronic acid (50% w/w), 17mg (15. mu. mol) of tetrakis (triphenylphosphine) palladium (0). The mixture was heated to reflux for 18 h. The warm mixture was filtered through Celite and the solvent removed in vacuo. The mixture was poured into water and extracted with dichloromethane. The organic layer was dried over sodium sulfate and evaporated. The residue was purified by HPLC to give 29mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.20(3H),1.59-1.88(2H),2.12(1H),3.48(2H),4.24(1H),4.40(1H),5.31(1H),7.04(1H),7.33(1H),7.47(1H),7.99(1H),8.12-8.21(2H),8.27(1H).

LC-MS (method 3) R_t＝1.27min；MS(ESIpos)m/z＝190[M+H]⁺⁺.

Example 42

(2R) -1- ({3- [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine

To a stirred solution of 100mg (0.37mmol) of (2R) -1- [ (3-bromoimidazo [1,2-b ] pyridazin-6-yl) oxy ] propan-2-amine in 6mL 1-propanol were added 550. mu.L (1.1mmol) of a 2M solution of potassium carbonate, 344mg (0.74mmol) of the crude [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] boronic acid (50% w/w), 17mg (15. mu. mol) of tetrakis (triphenylphosphine) palladium (0). The mixture was heated to reflux for 18 h. The warm mixture was filtered through Celite and the solvent removed in vacuo. The mixture was poured into water and extracted with dichloromethane. The organic layer was dried over sodium sulfate and evaporated. The residue was purified by HPLC to give 32mg of the title compound as a solid substance.

¹H-NMR(300MHz,DMSO-d₆),[ppm]＝1.20(3H),1.59-1.88(2H),2.12(2H),3.48(2H),4.24(1H),4.40(1H),5.31(1H),7.04(1H),7.33(1H),7.47(1H),7.99(1H),8.12-8.21(2H),8.27(1H)

LC-MS (method 3) R_t＝1.27min；MS(ESIpos)m/z＝190[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 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 a pharmaceutically acceptable carrier 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 (ophthalmic), sublingual, rectal, vaginal administration 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 with conventional tablet bases (e.g., lactose, sucrose, and corn starch) and in combination with: binders such as acacia, corn starch or gelatin, disintegrating agents such as potato starch, alginic acid, corn starch and guar gum, tragacanth, acacia for assisting the disintegration and dissolution of the tablets after administration, lubricants such as talc, stearic acid or magnesium stearate, calcium stearate or zinc stearate for improving the flowability of the tablet granulation and preventing adhesion of the tablet materials to the surfaces of the tablet dies and punches, dyes, colorants and flavouring agents such as peppermint, oil of wintergreen or cherry flavouring for improving the organoleptic properties of the tablets and making them more acceptable to the patient. Suitable excipients for oral liquid dosage forms include dicalcium phosphate and diluents such as water and alcohols (e.g., ethanol, benzyl alcohol, and polyvinyl alcohol), 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 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 include one or more preservatives, such as ethyl or n-propyl paraben; 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 injectable doses of the compounds, 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), 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 monoleate, and high molecular weight adducts of ethylene oxide with hydrophobic bases 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 polyethylene 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 less irritating fixed oil may be used, 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 to 1991, published on 6/11, 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," PDA Journal of Pharmaceutical Science & Technology 1997,51(4), 166-.

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, 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, glycerol 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 (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 min.

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 capsule: a large number of unit capsules were prepared by filling standard two-piece hard capsules with 100mg of powdered active ingredient, 150mg of lactose, 50mg of cellulose and 6mg of magnesium stearate, respectively.

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 units are taken orally without water for immediate dissolution and delivery of the drug. Mixing said active ingredients in a mixture comprisingSuch 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 agent" includes, but is not limited to: 131I-chTNT, abarelix, abiraterone, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, aminoglutethimide, amsacrine, anastrozole, Arglabin, arsenic trioxide, asparaginase, azacitidine, basiliximab, BAY 80-6946, BAY 1000394, BAY 86-9766(RDEA 119, belotecan, bendamustine, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, bortezomib, buserelin, cabazitaxel, calcium folinate, calcium levofolinate, capecitabine, carboplatin, carmofluorine, carmustine, saracaxomab, celecoxib, cimetimikin, cetuximab, bepotastine, chlorambucil, clorabeprinosine, clorpe, clorprenaline, clorpamide, clorprenaloside, atrazine, araffine, alfa, amazide, bax, bazedoxifene, bazedoary, clofarabine, clofarabi, Cytarabine, dacarbazine, dactinomycin, daltepatin alpha, dasatinib, daunorubicin, decitabine, degarelix, dinil interleukin fusion 2 toxin, dinolizumab, deslorelin, dibromospiro-chloride, docetaxel, doxifluridine, doxorubicin + estrone, eculizumab, eletrocumab, etiracetam, eltrompag, endostatin, enocitabine, epirubicin, epithiandrol, alfapentin, betanepatin, etaplatin, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, fadrozole, filgrastimastin, fludarabine, fluorouracil, flutamide, formestan, fulvestrant, gallium nitrate, ganix, gefitinib, gemcitabine, giemlugging, gefitinib, histremulin, histamine, doxycycline, i-125 seed, ibandronic acid, temozolomide, idarubicin, ifosfamide, imatinib, imiquimod, inpropulfan, interferon alpha, interferon beta, interferon gamma, ipilimumab, irinotecan, ixabepilone, lanreotide, lapatinib, lenalidomide, lenogrin, lentinan, letrozole, leuprorelin, levamisole, lisuride, lobaplatin, lomustine, lonidamine, maoprofol, medroxyprogesterone, megestrol, melphalan, mertansine, mercaptopurine, methotrexate, methoxsalen, aminolevulinic acid methyl ester, methyltestosterone, mifamurtide, miltefosine, mifuwhat, mitoxantrone, nelarabine, nilotinib, nimotuzumab, mukurozoxantrone, mukurozepine, mazine, mazotocin, niflumimazine, mazine, oxpocetine, mazine, gefitinib, orveduloxetine, gefitinib, and oxpocetine, Oxaliplatin, p53 gene therapy, paclitaxel, palifermin, palladium-103 seed, pamidronic acid, pamitumumab, pazopanib, pemetrexed, PEG-betaepoetin (methoxy PEG-betapotein, pegylated filgrastim, pegylated interferon alpha-2 b, pemetrexed, pentazocine, pentostatin, pellomycin, perphosphoramide, streptolysin, pirarubicin, plerixafor, priomycin, chitosan, polyetradiol polyphosphate, polysaccharose-k, porfimer sodium, pralatrexate, prednimustine, procarbazine, quinagolide, raloxifene, ranitrexed, ranimustine, propyleneimine, regorafenib, risedronate, rituximab, romidepsin, romiplosidasatin, sarsassertraline, sipuucel-T, levofelodil, tebuconazole, glycitin, streptozocin, sorafenib, sorafe, Talaporfin, tamibarotene, tamoxifen, tasolomine, tesil, tegafur + gimeracil + oteracil, temoporfin, temozolomide, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thioguanine, tosubuzumab, topotecan, toremifene, tositumomab, trabeutidine, trastuzumab, troosulfan, tretinoin, troostine, triptorelin, trofosfamide, tryptophan, ubenis, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vorinostat, vorolconazole, yttrium-90 glass microspheres, neat-statin, stastin ester, zoledronic acid, zotocin, or combinations thereof.

The additional agent may be afinitor, aldesleukin, alendronic acid, alpha-interferon (alfaferone), alitretinoin, allopurinol, sodium allopurinol for injection (alprimem), palonosetron hydrochloride injection (aloxi), altretamine, aminoglutethimide, amifostine, amsacrine, anastrozole, dolasetron tablet (anzmet), alfa bepotastine injection (aranesp), arglabin, diarsene trioxide, exemestane tablet, 5-azacytidine, azathioprine, BAY 80-6946, BCG or tide BCG, amastatin, betamethasone acetate, betamethasone sodium phosphate, bexarotene, bleomycin sulfate, bromozomib, busulfan, calcitonin, adalimumab (caath), capecitabine, cladribine, bivalinate, cline, melphalan, Clodronic acid, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin liposome (Daunoxome), dexamethasone sodium phosphate, estradiol valerate, dinierein 2(denileukin diftox), methylprednisolone, deslorelin, dexrazoxane, diethylstilbestrol, fluconazole, docetaxel, doxifluridine, doxorubicin, dronabinol, DW-166HC, leuprolide acetate (eligard), labyrinase injection (elitek), epirubicin hydrochloride injection (ellence), aprepirubicin capsule (emide), epirubicin, alfa eptin alfa, alfa fariptin (epigen), etaplatinum, levamisole, estradiol (estrrace), estradiol, estramustine sodium phosphate, ethisterol, amifostine, etidronic acid, etoposide, doxepirubicin, favudine, fafavudine, trofavudine, Finasteride, filgrastim, floxuridine, fluconazole, fludarabine, 5-fluorodeoxyuridine monophosphate, 5-fluorouracil (5-FU), fluoxymesterone, flutamide, formestane, fosetabine, fulvestrant, gamma-globulin (gammagard), gemcitabine, gemumab, imatinib mesylate (gleevec), carmustine wafer capsule (gliadel), goserelin, granisetron hydrochloride, histrelin, topotecan (hycamtin), hydrocortisone, erythroxylynoneadenine (eyrthro-hydroxyynyladine), 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, alpha-2A 2 interferon, alpha-n, Interferon alpha (intron A), gefitinib tablet (iressa), irinotecan, granisetron, lapatinib, lentinan sulfate, letrozole, leucovorin, leuprorelin acetate, levamisole, calcium levofolinate (levofolinic acid calcium salt), levothyroxine sodium (levothroid), levothyroxine sodium (levoxyl), lomustine, lonidamine, dronabinol, mechlorethamine, mecobalamin acetate, megestrol acetate, melphalan, esterified estrogen tablet (menest), 6-mercaptopurine, mesna, methotrexate, metivoreq, miltefosine, minocycline, mitomycin C, mitotane, mitoxantrone, trilostane (Modrenal), Myocoet, nedaplatin, filgrastimsin (neurin), recombinant interleukin 11 (neugeuma), glutethione (neugexenia), leucinogen (631570), milbevacene, milbemycin C, medetoposide, medetorphine (Modrenol), medetorphine, med, OCT-43, octreotide, ondansetron hydrochloride, cefixime (orapired), oxaliplatin, paclitaxel, prednisone sodium phosphate (pediapared), pemetrexed, pyroxene, pentostatin, streptolysin (pisibanil), pilocarpine hydrochloride, pirarubicin, plicamycin, porfimer sodium, prednimustine, prednisolone, prednisone, equine estrogen, 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 (salagen), octreotide, sargrastim, semustine, Sizopyran, sobuzosin, prednisolone, fosetyl, dry cell therapy, streptozocin, strontium chloride 89, streptozocin, levofloxacin, thyroxine, levofloxacin, and tamoxifen, Tamsulosin, tasolinamine, testolactone, docetaxel injection (taxotere), temocillin, temozolomide, teniposide, testosterone propionate, methyltestosterone, 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, dexrazoxane, stastin (zinostatin stimalamer), ondansetron, ABI-007, acolbifene, interferon gamma-1 b (actanimine), affinipak, aminopterin, acystus, apine, astrotrichum, astrotrichu, 9006 (B-6-S) (B-S6-B, S-S, S-D-E, S-D-E, S-D-E, S-D-S-E, S-, Atorvastatin (Avastin), CCI-779, CDC-501, celecoxib, cetuximab, clinacatto, cyproterone acetate, decitabine, DN-101, doxorubicin-MTC, dSLIM, dutasteride, edotararin, efluoromithine, efaprotecan, fenretinide, histamine dihydrochloride, histrelin hydrogel implants, holmium-166 DOTMP, ibandronic acid, gamma interferon, pegylated interferon alpha-2 b (intron-PEG), ixabepilone (ixabepilone), keyhole limpet hemocyanin (keyholeletpet hemocyanin), L-651582, lanreotide, lasofoxifene, libra, farnesol protein transferase inhibitors (lonanmnib), mirepredfene, minodronate (minodronate), MS-209, MTP-6, riluzole-6, lanreolin, neomycin, minoxidin, minophen S, meretrix, medrycin, mellitorine, and mefenone, osidmem, polyglutamic acid paclitaxel, pamidronate disodium, PN-401, QS-21, quazepam, R-1549, raloxifene, ranpirnase, 13-cis-retinoic acid, satraplatin, seocalcitol, T-138067, erlotinib hydrochloride tablet (tarceva), taxoprexin, alpha-1 thymosin, thifluzaline, tipifarnib, tirapazamine, TLK-286, toremifene, TransMID-107R, selevada, 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 by reference), such as asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, asparaginase, 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 in Goodmann and Gilman's The pharmaceutical basic of Therapeutics (9 th edition), edited by Molinoff et al, McGraw-Hill, pages 1225-1287 (1996) (incorporated by reference) for use in The treatment of neoplastic disease, such as aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine, cladribine, busulfan, diethylstilbestrol, 2' -difluorodeoxycytidine, docetaxel, erythrononyladenine, ethisterone, 5-fluorodeoxyuridine monophosphate, fludarabine phosphate, fluoxymesterone, flutamide, hydroxyprogesterone caproate, idarubicin, interferon, medroxyprogesterone acetate, megestrol acetate, melphalan, mitotane, 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), radiolabeled pertuzumab (Labetuzumab), radioisotope-crosslinked trastuzumab of eming particles, EM-Acuk-1421, interleukin (Hitachi), HPV-7, HPV-3516, HPV-30625, and, Javelin-melanoma, NY-ESO-1 vaccine, EGF vaccine, CYT-004-MelQbG10, WT1 peptide, agovacizumab (oregomab), ofatumumab, zalutumumab (zalutumumab), betheumatin interleukin (cindrekin bestudox), WX-G250, Albuferon, aflibercept, denosumab (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., atorvastatin, Rituxan, Erbitux, Herceptin) and recombinant proteins.

-one or more compounds of general formula (I) above, 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 agents selected from: a taxane, such as docetaxel, paclitaxel, lapatinib, sunitinib, or taxol; epothilones such as ixabepilone, paclitaxel (Patupilone) or salgopilone (Sagopilone); mitoxantrone; prednisolone; dexamethasone; estramustine; vinblastine; vincristine; 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 atorvastatin, Axitinib, DAST, recentin, sorafenib or sunitinib). It can also be combined with proteasome inhibitors, mTOR inhibitors, anti-hormones or steroid metabolic enzyme inhibitors.

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, treatment of 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.

Thus, the 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 methods 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, 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 sensitizes 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 yet 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 metastases thereof, such as leukemias and myelodysplastic syndromes, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, systemic cancers, and/or other diseases where no control of cell growth, proliferation and/or survival, inappropriate cellular immune, 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), 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 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.

The diseases mentioned in the first two 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 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, 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, 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 distant metastases. Such 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 variant), 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 disorders 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.

Reference throughout this document to the use of the term "treating" is conventional, e.g., for the purpose of counteracting, alleviating, reducing, alleviating, ameliorating the condition of a disease or disorder such as sarcoma, and the like.

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 growth of unrelated (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, 1996,37,855], neovascular glaucoma, psoriasis, retrolental 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 will be one to three times daily dosing to once every four weeks. 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 1500mg 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. Thus, the desired therapeutic regimen and the amount of a compound of the invention, pharmaceutically acceptable salt, ester or composition thereof to be administered 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 illustrate 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 the selected bioassay. When tested more than once, the data is reported as a mean or median value, where:

the mean, also called arithmetic mean, represents the total number of values obtained divided by the number of times tested, and

the median value represents the median of the set of values of the digits 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 MKNK1 TR-FRET assay as described in the following section.

Recombinant fusion proteins of glutathione-S-transferase (GST, N-terminal) 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 Sepharose affinity chromatography were purchased from Carna Biosciences (product No. 02-145) 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 MKNK 1in 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. The kinase reaction was then 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 incubating the resulting mixture 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 adding 5. mu.L of a solution of TR-FRET detection reagent (5nM streptavidin-XL 665[ Cisbio Bioassays, Codolet, France ] and 1nM anti-ribosomal protein S6(pSer236) -antibody from Invitrogen [ #44921G ] and 1 nMLLANCE EU-W1024-labeled protein G [ Perkin-Elmer, product number AD0071] in aqueous EDTA (100mMEDTA, 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 emission at 620nm and 665nm after excitation at 350nm is measured using a TR-FRET reader, for example (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 on the same microtiter plate, two values are tested for each concentration, and IC is calculated using 4-parameter fitting₅₀Values, the concentration 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 and0.1nM, prepared separately as a 1:3.4 dilution series at a 100-fold level of concentrated DMSO solution prior to assay).

Table 1: MKNK1 IC₅₀

Examples	MKNK1 IC50[nM]
		1	17
10	34
		11	17
12	26
		13	5
2	3
		3	11
4	20
		5	21
6	23
		7	25
8	28
		9	48
14	5
		15	12
16	8
		17	67
18	17
		19	17
20	64
		21	10
22	14
		23	32
24	3
		25	16
26	100
		27	153
28	7
		29	8

30	137
		31	13
32	14
		33	99
34	5
		35	21
36	4
		37	7
38	8
		39	250
40	5

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 a TR-FRET based MKNK1 high ATP assay as described in the following section.

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 MKNK 1in 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. The kinase reaction was then 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 adding 5. mu.L of a solution of TR-FRET detection reagent (5nM streptavidin-XL 665[ Cisbio Bioassays, Codolet, France ] and 1nM anti-ribosomal protein S6(pSer236) -antibody from Invitrogen [ #44921G ] and 1 nMLLANCE EU-W1024-labeled protein G [ Perkin-Elmer, product number AD0071] in aqueous EDTA (100mMEDTA, 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 (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 on the same microtiter plate, two values are tested for each concentration, and IC is calculated using 4-parameter fitting₅₀Values, stated at concentrations 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, respectively, were prepared by serial dilution at 100-fold the level of concentrated DMSO solution prior to assayThe exact concentration may vary depending on the pipette used in preparation for the dilution series).

Table 2: MKNK1 high ATP IC₅₀

Examples	MKNK1 high ATP IC50[ nM]
		1	53
10	62
		11	59
12	65
		13	16
2	5
		3	27
4	91

5	65
		6	37
7	91
		8	136
9	94
		14	34
15	31
		16	19
17	127
		18	43
19	34
		20	132
21	31
		22	79
23	57
		24	4
25	28
		26	216
27	259
		28	13
29	16
		30	309
31	28
		32	37
33	117
		34	15
35	44
		36	4
37	17
		38	23
39	593

40	13
		41	19
42	26

CDK2/CycE kinase assay

The CDK 2/CycE-inhibitory activity of the compounds of the invention was quantified using the CDK2/CycE TR-FRET assay as described in the following section.

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 the 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 ph8.0,10mM magnesium chloride, 1.0mM dithiothreitol, 0.1mM sodium orthovanadate, 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. The kinase reaction was then 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 incubating the resulting mixture 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 adding 5. mu.L of TR-FRET detection reagent (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 AD0077, as an alternative, a terbium-cryptate-labeled anti-mouse IgG antibody from Cisbio Bioassays ] in aqueous EDTA (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, a TR-FRET reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (R) ((R))Perkin-Elmer) to measure 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 11 different concentrations on the same microtiter plate, two values are tested for each concentration, and IC is calculated using 4-parameter fitting₅₀Values, the concentration was 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 prepared separately by 1:3.4 serial dilutions at 100-fold the level of concentrated DMSO solution prior to assay).

PDGFR beta kinase assay

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

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/naohph7.5,10mM 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. The kinase reaction was then 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 incubating the resulting mixture at 22 ℃ for a reaction time of 25 min. The PDGFR β concentration in the assay was adjusted according to enzyme batch activity and was chosen appropriately to bring the assay in the linear range, with typical enzyme concentrations ranging from 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 (200nM streptavidin-XLent [ CisBioInternational ] and 1.4nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Tb-chelate from CisBioInternational can also be used instead of PT 66-Eu-chelate ]) in a solution of aqueous EDTA (100mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES/NaOH pH 7.5).

The reaction 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 assessed 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 using 4-parameter fitting₅₀Values, the concentration was 20. mu.M to 0.1nM (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 prepared separately by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

Fyn kinase assay

The His 6-tagged human recombinant kinase domain at the C-terminus of human T-Fyn was used as kinase, which was expressed in baculovirus-infected insect cells (purchased from Invitrogen, P3042). The biotinylated peptide biotin-KVEKIGEGTYGVV (C-terminus 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 T-Fyn was added in aqueous assay buffer [25 mM Tris/hci ph 7.2,25mM 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 before starting the kinase reaction. The kinase reaction was then 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 a solution of HTRF detection reagent (0.2. mu.M streptavidin-XL [ Cisbio Bioassays, Codolet, France ] and 0.66 nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Tb-cryptate from Cisbio Bioassays can also be used instead of PT 66-Eu-chelate ]) in aqueous EDTA solution (125 mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES/NaOH pH 7.0).

The reaction mixture was incubated at 22 ℃ for 1h to bind 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, 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, the test compounds are present in 10 different microtiter platesTest at different concentrations, test two values per concentration, and calculate IC using fitting by 4 parameters₅₀Values, the concentration was 20. mu.M to 0.1nM (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 prepared separately by 1:3 serial dilutions at the level of 100-fold concentrated stock solution prior to assay).

Flt4 kinase assay

Flt4 TR-FRET assays, as described in the following sections, were used to quantify Flt4 inhibitory activity of 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 uL of Flt4 in aqueous assay buffer [25 mM HEPES pH7.5,10mM magnesium chloride, 2mM dithiothreitol, 0.01% (v/v) Triton-X100(Sigma),0.5 mM EGTA and 5mM β -glycerophosphate ] was added, and the mixture was incubated at 22 ℃ for 15min to pre-bind the test compound to the enzyme prior to the start of the kinase reaction. The kinase reaction was then 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, typical enzyme concentrations were 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. mu.L of HTRF detection reagent (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 (50 mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES pH 7.5).

The reaction 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 (BMGLAbtechnologies, 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 using 4-parameter fitting₅₀Values, the concentration was 20. mu.M to 0.1nM (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 prepared separately 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 section.

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 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 ] 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,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 being 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 (30nM streptavidin-XL 665[ Cis Biointernational ] and 1.4nM PT 66-Eu-chelate (europium-chelate labeled anti-phosphotyrosine antibody from Perkin Elmer [ PT 66-Tb-cryptate from Cis Biointernational can also be used instead of PT 66-Eu-chelate ]) in aqueous EDTA solution (100mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES/NaOH pH 7.5).

The reaction 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 evaluated by measuring the resonance energy transfer from PT 66-Eu-chelate 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 using 4-parameter fitting₅₀Values, said concentration being from 20. mu.M to 0.1nM (20)μ M, 6.7 μ M, 2.2 μ M, 0.74 μ M, 0.25 μ M, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, the dilution series being prepared separately by 1:3 serial dilutions at 100-fold levels of 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 technique allows for the determination 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 a unimodal oxygen atom, which triggers 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-eIF4ESer20910K assay kit and the AlphaScreen protein A kit (for 10K assay spots), both from Perkin Elmer, were used.

On the first day, 50.000A 549 cells in growth medium (DMEM/Hams' F12 with stabilized glutamine, 10% FCS) were seeded at 100. mu.L per well in 96-well plates 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 serially diluted in 50 μ L starvation medium and final DMSO concentration was 1% and added to a549 cells in the test plate at a final concentration range of 10 μ M up to 10nM low depending on the concentration of test compound. The treated cells were incubated at 37 ℃ for 2 h. Add 37 μ l FCS to wells (20% final FCS concentration) for 20 min. 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 (proxoplate, from PerkinElmer) 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 an additional 2h at room temperature with gentle shaking. The plates were then measured in an EnVision reader (PerkinElmer) with AlphaScreen program. Each data point (compound dilution) was measured in triplicate.

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

It will be apparent to those skilled in the art that other MKNK-1 kinase assays may be similarly performed using suitable reagents.

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 metastases thereof, such as leukemias and myelodysplastic syndromes, malignant lymphomas, head and neck tumors including brain tumors and brain metastases, tumors, cancers of the head and neck, and the like, 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 formula (I), or a stereoisomer or a salt thereof, or a mixture thereof:

wherein:

represents a group selected from:

C₁-C₆-alkyl-, 3-to 6-membered heterocycloalkyl connected as a spiro ring, C₆-C₁₄-aryl-, C₆-C₁₄-aryl-C₁-C₆-alkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, -NH₂、-NHR’、-N(R’)R”、-N(H)C(＝O)OR’、-N(R’)C(＝O)OR’、-OH、C₁-C₆-alkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-alkoxy-, -SH, C₁-C₆-alkyl-S-;

r4 represents a hydrogen atom;

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

C₁-C₆-alkyl-;

n represents an integer of 0 or 1.

2. The compound of claim 1, or a stereoisomer or salt thereof, or a mixture thereof, wherein:

represents a group selected from:

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

halogen atom, -CN, C₁-C₆-alkyl-, -NHR', -OH, C₁-C₆-alkoxy-, C₃-C₆-cycloalkoxy-, C₃-C₆-cycloalkyl-C₁-C₃-an alkoxy-group;

r4 represents a hydrogen atom;

r' represents a substituent selected from:

C₁-C₆-alkyl-;

n represents an integer of 0 or 1.

3. The compound of claim 1 or 2, or a stereoisomer or salt thereof, or a mixture thereof, wherein:

represents a group selected from:

r1 represents optionally substituted one or moreStraight chain C substituted with one substituent independently selected from₂-C₆Alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-a cycloalkyl group:

3-to 6-membered heterocycloalkyl connected as a spiro ring, C₆-C₁₄-aryl-, C₆-C₁₄-aryl-C₁-C₆-alkoxy-;

r2 represents a hydrogen atom;

r3 represents a substituent selected from:

r4 represents a hydrogen atom;

n represents an integer of 0 or 1.

4. The compound of claim 1 or 2 selected from:

(1S) -2- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine;

trans-3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine;

(2R) -1- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

(1S) -2- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine;

(2S) -1- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

(2R) -2- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine;

(1R) -2- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine;

(2R) -2- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-amine;

(1R) -2- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -1-phenethylamine;

(2R,3R) -3- (benzyloxy) -1- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } butan-2-amine;

(2R) -1- (benzyloxy) -3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

(2S) -1- { [3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

trans-3- { [3- (furo [2,3-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine formate salt;

trans-3- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine formate salt;

(2R) -2-amino-3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-1-ol;

3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-methylpropan-1-amine;

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

(1S,3R) -3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclopentylamine;

(2S) -1- ({3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine;

(2S) -1- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

trans-4- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclohexanecarboxylate;

1- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-methylpropan-2-amine;

3- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -2-phenylpropan-1-amine;

trans-3- ({3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) cyclobutylamine;

(2S) -1- ({3- [4- (cyclopropylmethoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine;

(2R) -1- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

1- [3- ({ [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } methyl) oxetan-3-yl ] methylamine;

trans-3- { [3- (furo [3,2-b ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine;

trans-3- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine;

(2S) -1- { [3- (furo [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -3-methylbutan-2-amine;

(1S,2S) -2- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclopentylamine;

(2S) -1- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

2- { [3- (4-methoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } -3-phenylpropan-1-amine;

(2S) -1- ({3- [4- (2, 2-dimethylpropoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine;

2- {6- [ (trans-3-aminocyclobutyl) oxy ] imidazo [1,2-b ] pyridazin-3-yl } furo [3,2-c ] pyridin-4-ol;

trans-3- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } cyclobutylamine;

trans-3- ({3- [4- (prop-2-yloxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) cyclobutylamine;

(2R) -1- { [3- (4-ethoxyfuro [3,2-c ] pyridin-2-yl) imidazo [1,2-b ] pyridazin-6-yl ] oxy } propan-2-amine;

tert-butyl [2- (6- { [ (2S) -2-aminopropyl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) furo [3,2-c ] pyridin-4-yl ] ethylcarbamate;

2- (6- { [ (2S) -2-aminopropyl ] oxy } imidazo [1,2-b ] pyridazin-3-yl) -N-ethylfuro [3,2-c ] pyridin-4-amine;

(2S) -1- ({3- [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine; and

(2R) -1- ({3- [4- (cyclobutoxy) furo [3,2-c ] pyridin-2-yl ] imidazo [1,2-b ] pyridazin-6-yl } oxy) propan-2-amine.

5. A process for the preparation of a compound of general formula (I) according to any one of claims 1 to 4, comprising the following steps: reacting an intermediate compound of formula (V):

wherein A, R2, R3, R4 and n are as defined for a compound of general formula (I) according to any one of claims 1 to 4, and X represents a leaving group which is a halogen atom or a perfluoroalkylsulfonate group,

with a compound of the general formula (III):

wherein R1 is as defined for a compound of general formula (I) according to any one of claims 1 to 4,

thereby obtaining a compound of formula (I):

6. the process according to claim 5, wherein the halogen atom is a chlorine, bromine or iodine atom and the perfluoroalkylsulfonate group is a trifluoromethylsulfonate group or a nonafluorobutylsulfonate group.

7. A pharmaceutical composition comprising a compound of general formula (I), or a stereoisomer or a salt thereof, or a mixture thereof, according to any one of claims 1 to 4, and a pharmaceutically acceptable diluent or carrier.

8. The pharmaceutical composition of claim 7, wherein the salt is a pharmaceutically acceptable salt.

9. A pharmaceutical composition comprising:

-one or more first active ingredients selected from compounds of general formula (I) according to any one of claims 1 to 4, 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 or a salt thereof, or a mixture thereof, according to any one of claims 1 to 4, for the preparation of a medicament for the prevention or treatment of a disease resulting from uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response mediated by the MKNK-1 pathway.

11. The use of claim 10, wherein the disease caused by uncontrolled cell growth, proliferation and/or survival, an inappropriate cellular immune response or an inappropriate cellular inflammatory response is a hematological tumor, a solid tumor and/or metastases thereof.

12. Use according to claim 11, wherein the hematological tumors, solid tumors and/or metastases thereof are selected from the group consisting of leukemias and myelodysplastic syndromes, malignant lymphomas, tumors of the head and neck, of the thorax, of the gastrointestinal tract, of endocrine tumors, of the breast and other gynaecological tumors, of the urinary system, of skin tumors and sarcomas, and/or metastases thereof.

13. The use according to claim 12, wherein the head and neck tumor is selected from the group consisting of brain tumors and brain metastases.

14. The use of claim 12, wherein the breast tumor is selected from the group consisting of a non-small cell lung tumor and a small cell lung tumor.

15. The use of claim 12, wherein the urological tumour is selected from the group consisting of renal tumours, bladder tumours and prostate tumours.

16. Use of a compound of formula (V) for the preparation of a compound of formula (I) according to any one of claims 1 to 4:

wherein A, R2, R3, R4 and n are as defined for the compound of general formula (I) according to any one of claims 1 to 4, and X represents a leaving group which is a halogen atom or a perfluoroalkylsulfonate group.