HK1194067A

HK1194067A - Thiazole derivatives

Info

Publication number: HK1194067A
Application number: HK14107444.1A
Authority: HK
Inventors: S.R.卡拉; W．斯戴勒; E．斯道布; M．乌赫勒-普列特克
Original assignee: 默克专利有限公司
Priority date: 2011-05-23
Filing date: 2012-04-12
Publication date: 2014-10-10

Description

Thiazole derivatives

Technical Field

The object of the present invention was to find novel compounds with valuable properties, in particular compounds which can be used for the preparation of medicaments.

The present invention relates to pyridine compounds capable of inhibiting one or more kinases. These compounds are useful in the treatment of a variety of diseases including cancer, septic shock, Primary Open Angle Glaucoma (POAG), hyperplasia, rheumatoid arthritis, psoriasis, atherosclerosis, retinopathy, osteoarthritis, endometriosis, chronic inflammation, and/or neurodegenerative diseases such as alzheimer's disease.

The present invention relates to compounds and their use in the inhibition, control and/or modulation of signal transduction by kinases, especially receptor tyrosine kinases, to pharmaceutical compositions comprising these compounds and to the use of said compounds in the treatment of kinase-induced diseases.

Background

Protein kinases are ideal targets for therapeutic intervention in a variety of disease states, as they regulate almost every cellular process, including metabolism, cell proliferation, cell differentiation, and cell survival. For example, protein kinases play a key role in cell cycle control and angiogenesis, both of which are associated with many diseases such as, but not limited to, cancer, inflammatory diseases, abnormal angiogenesis and its related diseases, atherosclerosis, macular degeneration, diabetes, obesity, and pain.

The invention relates inter alia to compounds in which inhibition, control and/or modulation of signaling of TBK1 and IKK epsilon is of interest, and uses thereof.

One of the basic mechanisms for performing cell regulation is the conduction of extracellular signals across the membrane, which in turn regulate biochemical processes within the cell. Protein phosphorylation represents a process by which intracellular signals are conducted between molecules, which ultimately leads to the generation of a cellular response. These signaling cascades are highly regulated and often overlap, as evidenced by the presence of many protein kinases as well as phosphatases. Phosphorylation of proteins occurs predominantly at serine, threonine or tyrosine residues, and protein kinases are therefore classified by the specificity of their phosphorylation sites, i.e., serine/threonine kinases and tyrosine kinases. Since phosphorylation is a ubiquitous process in cells, and cellular phenotype is highly influenced by channel activity, it is currently believed that many disease states and/or diseases are attributable to aberrant activation or functional variation of the molecular components of the kinase cascade. Therefore, there is considerable interest in the characterization of these proteins and compounds capable of modulating their activity (for review, Weinstein-Oppenheimer et al Pharma. &. Therap.,2000,88, 229-279).

IKK epsilon and TBK1 are serine/threonine kinases that are highly homologous to each other and to other IkB kinases. Both kinases play an important role in the innate immune system. Toll-like receptors 3 and 4 and RNA helicases RIG-I and MDA-5 recognize double stranded RNA viruses, resulting in activation of the TRIG-TBK1/IKK ε -IRF3 signaling cascade and resulting in a type I interferon response.

In 2007, Boehm et al described IKK epsilon as a novel oncogene in breast cancer [ j.s. Boehm et al, cell129,1065-1079,2007 ]. The ability of 354 kinases to reproduce the Ras transformation phenotype when used in combination with the activated form of the MAPK kinase Mek was studied. Among them, IKK epsilon is identified as a synergistic oncogene. In addition, the authors demonstrated that IKK epsilon is amplified and overexpressed in a number of breast cancer cell lines and tumor samples. Apoptosis is induced and proliferation is hindered by the reduction of gene expression by RNA interference in breast cancer cells. Similar findings were obtained by Eddy et al 2005, which highlighted the importance of IKK ε in breast Cancer disease [ S.F.Eddy et al, Cancer Res.2005;65(24),11375-11383 ].

The cancer promotion effect of TBK1 was first reported in 2006. In screening a gene bank containing 251,000 cdnas, Korherr et al accurately identified three genes involved in innate immune defense as pro-angiogenic factors: TRIF, TBK1 and IRF3[ C.Korherr et al, PNAS,103, 4240-. Chien et al [ Y.Chien et al, Cell127,157-170,2006] reported that TBK-/-cells could only be transformed to a limited extent using oncogenic Ras in 2006, indicating that TBK1 is involved in Ras-mediated transformation. In addition, they also demonstrated that RNAi-mediated low levels of TBK1 trigger apoptosis in MCF-7 and Panc-1 cells. Barbie et al recently reported that TBK1 has great importance in many cancer cell lines with variant K-Ras, suggesting that TBK1 intervention is of importance in the treatment of related tumors [ D.A. Barbie et al, NatureLetters1-5,2009 ].

Diseases caused by protein kinases are characterized by abnormal activity or overactivity of the protein kinase. The aberrant activity involves: (1) expression in cells that do not normally express these protein kinases; (2) enhanced kinase expression, which results in undesirable cell proliferation, such as cancer; or (3) enhanced kinase activity, which results in undesirable cell proliferation such as cancer and/or overactivity of the corresponding protein kinase. Overactivity involves the amplification of a gene encoding a protein kinase, or the generation of activity levels associated with a cell proliferative disorder (i.e., one or more symptoms of the cell proliferative disorder increase in severity with increased kinase levels). The bioavailability of a protein kinase can also be affected by the presence or absence of a group of binding proteins for that kinase.

IKK epsilon and TBK1 are highly homologous serine/threonine kinases that are critically involved in innate immune responses induced by type 1 interferons and other cytokines. These kinases are stimulated in response to viral/bacterial infections. The immune response to viral and bacterial infections involves the binding of antigens such as bacterial Lipopolysaccharide (LPS), viral double stranded rna (dsrna) to Toll-like receptors, and the subsequent activation of the TBK1 pathway. The activated TBK1 and IKK epsilon phosphorylate IRF3 and IRF7, which triggers dimerization and nuclear translocation of these interferon regulated transcription factors, and finally induces a signaling cascade leading to IFN production.

More recently, IKK epsilon and TBK1 have also been shown to be associated with cancer. IKK epsilon has been shown to cooperate with activated MEK to transform human cells. In addition, IKK epsilon is often amplified/overexpressed in breast cancer cell lines and tumors from patients. TBK1, induced under hypoxic conditions, is expressed at significant levels in many solid tumors.

In addition, TBK1 is required to support oncogenic Ras transformation, whereas TBK1 kinase activity is elevated in transformed cells and TBK1 kinase activity is required for their survival in culture. Similarly, TBK1 and NF-kB signaling were also found to be extremely important in KRAS variant tumors. TBK1 has been identified as a synthetic lethal partner of oncogenic KRAS.

The literature:

y. H.Ou et al, Molecular cell41,458-470,2011;

a. Barbie et al, Nature,1-5,2009.

Accordingly, the compounds according to the invention, or pharmaceutically acceptable salts thereof, are useful for the treatment of cancer, including solid tumors, for example, tumors (e.g., lung, pancreatic, thyroid, bladder or colon cancer), myeloid disorders (e.g., myeloid leukemia), or adenomas (e.g., villous colon adenomas).

The tumor also includes monocytic leukemia, brain tumor, genitourinary tumor, lymphatic system tumor, gastric cancer, laryngeal and lung tumor (including lung adenocarcinoma and small cell lung cancer), pancreatic cancer and/or breast cancer.

The compounds are also useful for the treatment of HIV-1 (human immunodeficiency virus type 1) induced immunodeficiency.

By carcinomatous hyperproliferative diseases are understood brain cancer, lung cancer, squamous epithelial cancer, bladder cancer, gastric cancer, pancreatic cancer, liver cancer, kidney cancer, colorectal cancer, breast cancer, head tumor, neck tumor, esophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia and acute leukemia. In particular, cancer-like cell growth is a disease that represents the object of the present invention. The present invention therefore relates to compounds according to the invention for use as medicaments and/or pharmaceutical active ingredients for the treatment and/or prophylaxis of said diseases, to the use of compounds according to the invention for the preparation of medicaments for the treatment and/or prophylaxis of said diseases, and to methods for the treatment of said diseases, which methods comprise the administration of one or more compounds according to the invention to a patient in need thereof.

The compounds of the invention can be shown to have an antiproliferative effect in vivo in xenograft tumor models. The compounds of the invention can be administered to a subject suffering from a hyperproliferative disorder, for example, to inhibit tumor growth, reduce inflammation associated with lymphoproliferative disorders, inhibit graft rejection or nerve damage due to tissue repair. The compounds of the invention are suitable for prophylactic or therapeutic purposes. The term "treatment" as used herein refers to both prevention of a disease and treatment of an existing disease. Administration of the compounds of the invention before significant disease development can prevent proliferation/growth, e.g., prevent tumor growth, prevent metastatic tumor growth, reduce restenosis associated with cardiovascular surgery, etc. Alternatively, treatment of an existing disease with the compound may stabilize or improve the clinical symptoms of the patient.

The host or patient may be of a mammal, such as a primate, in particular a human; rodents, including mice, rats, and hamsters; rabbits, horses, cattle, dogs, cats, etc. Animal models of interest for experimental studies can provide models for the treatment of human diseases.

Sensitivity to treatment of a particular cell with a compound of the invention can be determined in vitro. Cell cultures are generally incubated with various concentrations of the compounds of the invention for a time sufficient for the active agent to induce cell death or inhibit migration, often between 1 hour and 1 week. In vitro assays can be performed using cultured cells from a biopsy sample. Viable cells remaining after treatment were then counted.

The dosage administered will vary depending on the particular compound used, the particular disease, the condition of the patient, etc. The therapeutic dose should generally be sufficient to reduce the undesirable cell population in the target tissue while maintaining patient survival. Such treatment is generally continued until the cancer cell load is reduced considerably, for example by at least 50%, and treatment is continued until substantially no more undesirable cells are detected in the body.

Many diseases are associated with dysregulation of cell proliferation and cell death (apoptosis). Diseases of interest include, but are not limited to, the following. The compounds of the invention are suitable for the treatment of various diseases in which smooth muscle cells and/or inflammatory cells proliferate and/or migrate into the intimal layer of blood vessels, resulting in restricted blood flow through the vessels, e.g., obstructing the neointimal vessels of the lesion. Graft vessel occlusive diseases of interest include: atherosclerosis, post coronary vascular graft disease, graft vein stenosis, restenosis around the anastomosis of vascular prostheses, restenosis following angioplasty or stenting, and the like.

Furthermore, the use of the compounds according to the invention may achieve additional or synergistic effects in and/or restore the efficacy of certain existing cancer chemotherapies and radiation therapies.

The term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures known to those skilled in the chemical, pharmaceutical, biological, biochemical and medical arts, and those manners, means, techniques and procedures resulting from simple modifications of such known manners, means, techniques and procedures by those skilled in the art.

The term "administering" as used herein refers to a method of contacting a compound of the invention with a target kinase in a manner such that the compound is capable of affecting the enzymatic activity of the kinase, either directly (i.e., interacting with the kinase itself) or indirectly (i.e., interacting with another molecule upon which the catalytic activity of the kinase is dependent). The administration described herein can be performed in vitro (i.e., in a test tube) or in vivo (i.e., in a cell or tissue of a living organism).

In the present specification, the term "treating" includes eliminating, substantially inhibiting, slowing or reversing the progression of the disease or disorder, substantially alleviating the clinical symptoms of the disease or disorder, or substantially preventing the appearance of the clinical symptoms of the disease or disorder.

In the present specification, the term "prevention" refers to a method of preventing a disease or disorder in an organism from the outset.

For any compound used in the present invention, a therapeutically effective amount, also referred to herein as a therapeutically effective dose, can first be estimated from cell culture assays. For example, doses can be formulated in animal models to obtain circulating concentration ranges including IC50 or IC100 determined in cell culture. This information can be used to more accurately determine the effective dose in humans. The starting dose can also be estimated from in vivo data. Using these starting guidelines, one skilled in the art can determine effective dosages in humans.

In addition, toxicity and therapeutic efficacy of the compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals (e.g., by determining LD50 and ED 50). The dose ratio between toxic and therapeutic effectiveness is the therapeutic index, which can be expressed as the ratio between LD50 and ED 50. Compounds with high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used to formulate a non-toxic dosage range for use in humans. Preferably, the dose of the compound is within a range of circulating concentrations that include ED50 with little or no toxicity. The dosage may vary within the stated ranges depending upon the dosage form employed and the route of administration. The exact formulation, route of administration and dosage can be selected by the individual physician in accordance with the patient's circumstances (see, e.g., Fingl et al, 1975, first page of the first chapter of the pharmacological Basis of Therapeutics).

The dosage and interval may be adjusted individually so that the concentration of the active compound in the plasma is sufficient to maintain the therapeutic effect. Typically, the dosage for oral administration to a patient will be in the range of about 50-2000 mg/kg/day, typically about 100-1000 mg/kg/day, preferably about 150-700 mg/kg/day, and most preferably about 250-500 mg/kg/day.

Preferably, therapeutically effective serum concentrations are obtained by multiple daily administrations. In the case of topical administration or selective ingestion, the effective local concentration of the drug may be independent of plasma concentration. Those skilled in the art will be able to optimize the local therapeutically effective dose without undue experimentation.

The compounds described in the present specification are preferably used for the prevention, treatment and/or study of cell proliferative diseases, especially cancer, such as, but not limited to: papilloma, glioma, kaposi's sarcoma, melanoma, lung cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, astrocytoma, head tumor, neck tumor, skin cancer, liver cancer, bladder cancer, breast cancer, lung cancer, uterine cancer, prostate cancer, testicular cancer, colorectal cancer, thyroid cancer, pancreatic cancer, gastric cancer, hepatocellular carcinoma, leukemia, lymphoma, hodgkin's disease, and burkitt's disease.

Prior Art

Other heterocyclic derivatives and their use as antitumor agents are described in WO 2007/129044.

The use of other pyridine and pyrazine derivatives in the treatment of cancer is described in WO2009/053737,

WO2004/055005 describes the use of these compounds in the treatment of other diseases.

Other heterocyclic derivatives are disclosed as IKK epsilon inhibitors in WO 2009/122180.

Pyrrolopyrimidines as inhibitors of IKK epsilon and TBK1 are described in WO 2010/100431.

Pyrimidine derivatives as IKK epsilon and TBK1 inhibitors are described in WO 2009/030890.

Disclosure of Invention

The invention relates to a compound shown in a general formula I,

in the formula (I), the compound is shown in the specification,

x represents H, CONH₂Or the CN group is selected from the group consisting of,

y represents NH, N-Me, S or O,

r represents Ar or Het in the presence of a catalyst,

R¹represents phenyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, pyridazinyl, indolyl, isoindolyl, benzimidazolyl, indazolyl, quinolyl, 1, 3-benzodioxolyl, benzothienyl, benzofuryl, imidazopyridinyl or furo [3,2-b ] group]Pyridyl, each of the above radicals being unsubstituted OR substituted by Hal, A, OR⁵、CN、COOA、COOH、CON(R⁵)₂And/or NR⁵The COA' is mono-or di-substituted,

ar represents phenyl, diphenyl or naphthyl, each of which is unsubstituted or substituted by Hal, A, Het¹、(CH₂)_nHet²、(CH₂)_nOR⁵、(CH₂)_nN(R⁵)₂、NO₂、CN、(CH₂)_nCOOR⁵、(CH₂)_nCON(R⁵)₂、CONH(CH₂)_qNHCOOA'、CON[R⁵(CH₂)_nHet¹]、NR⁵COA、NHCOOA、NR⁵SO₂A、COR⁵、SO₂Het²、SO₂N(R⁵)₂And/or S (O)_pA is mono-, di-or tri-substituted,

het represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, thiadiazole, pyridazinyl, pyrazinyl, indolyl, isoindolyl, benzimidazolyl, indazolyl, quinolyl, 1, 3-benzodioxolyl, benzothienyl, benzofuranyl or imidazopyridinyl, each of which is unsubstituted or substituted by A, COA, (CH)₂)_pHet¹、(CH₂)_pHet²、OH、OA、OAr、Hal、(CH₂)_pN(R⁵)₂、NO₂、CN、(CH₂)_pCOOR⁵、(CH₂)_pCON(R⁵)₂、NR⁵COA、(CH₂)_pCOHet²And/or (CH)₂)_pPhenyl is mono-, di-or tri-substituted,

Het¹represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, thiadiazole, pyridazinyl, pyrazinyl, each of which is unsubstituted or substituted by A, OH, OA, Hal, CN and/or (CH)₂)_pCOOR⁵Mono-, di-or tri-substituted,

Het²represents a dihydropyrrolyl group, a pyrrolidinyl group, a tetrahydroimidazolyl group, a dihydropyrazolyl group, a tetrahydropyrazolyl group, a dihydropyridinyl group, a tetrahydropyridinyl group, a piperidinyl group, a,Morpholinyl, hexahydropyridazinyl, hexahydropyrimidyl, [1, 3]]Dioxolanyl, piperazinyl, each of which is unsubstituted or substituted by OH, COOA', CON (R)⁵)₂The COA and/or the A monosubstitution,

a' represents a straight-chain or branched alkyl group having 1 to 6 carbon atoms, wherein 1 to 7 hydrogen atoms may be substituted by F,

a represents a linear or branched alkyl group containing 1 to 10 carbon atoms, wherein one or two non-adjacent CH and/or CH₂Which groups may be substituted by nitrogen, oxygen, sulfur atoms and/or by-CH = CH groups, and/or wherein 1-7 hydrogen atoms may be substituted by F,

R⁵represents H or a linear or branched alkyl group containing 1 to 6 carbon atoms, in which 1 to 7 hydrogen atoms may be substituted by F,

hal represents F, Cl, Br or I,

n represents 0,1, 2,3, 4or 5,

p represents 0,1 or 2,

q represents 1,2, 3 or 4,

and pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios.

The invention also relates to optically active forms (stereoisomers), salts, enantiomers, racemates, diastereomers and hydrates and solvates of these compounds. The term "solvate of a compound" is used to indicate an adduct of such a compound with an inert solvent molecule formed by the attractive forces of each other. Solvates are, for example, mono-or bis-hydrates or alcoholates. Of course, the invention also relates to solvates of the salts.

The term "pharmaceutically acceptable derivatives" is used to indicate, for example, salts and so-called prodrug compounds of the invention.

The term "prodrug derivative" is intended to denote compounds of the general formula I modified with, for example, alkyl or acyl groups, sugars or oligopeptides, which are rapidly cleaved in the body to form the active form of the compound.

Also included among these are polymeric derivatives of the biodegradable compounds of the present invention, see, for example, int.115,61-67 (1995).

The term "effective amount" means an amount of a drug or pharmaceutical active ingredient that will elicit the production of a biological or medical response of a tissue, system, animal or human that is, for example, sought or desired by a researcher or physician.

Furthermore, the term "therapeutically effective amount" means that the amount produces the following result compared to a corresponding patient who does not receive such amount:

treating improves, cures, prevents or eliminates a disease, syndrome, condition, patient complaint, illness, or side effect or also slows the progression of a disease, complaint, or illness.

The term "therapeutically effective amount" also encompasses an amount effective to enhance normal physiological function.

The invention also relates to the use of mixtures of compounds of the general formula I, for example mixtures of two diastereomers, for example in a mixing ratio of 1: 1,1: 2,1: 3,1: 4,1: 5,1: 10,1: 100 or 1: 1000.

these mixtures are particularly preferably mixtures of stereoisomeric compounds.

The invention also relates to compounds of the general formula I and salts thereof, and to a process for the preparation of compounds of the general formula I and pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, according to claims 1 to 12, which process is characterized in that,

a) a compound of the general formula II,

wherein X, Y and R¹Having the meaning given in claim 1,

reacting with a compound shown in a general formula III,

R-CO-L III

in which R has the meaning given in claim 1, L represents Cl, Br, I or a free OH group or a reactive, functionally modified OH group,

alternatively, the first and second electrodes may be,

b) by treating one of the functional group derivatives with a solvolytic or hydrogenolytic agent, thereby releasing the compound,

and/or converting a base or acid of formula I into a salt thereof.

In this context, the radical or the parameter R¹R and X have the same meanings as indicated under formula I, unless explicitly stated otherwise.

A represents an alkyl group, is unbranched (linear) or branched, and contains 1,2, 3,4,5,6, 7,8, 9 or 10 carbon atoms. A preferably represents methyl, and may also be ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, or may also be pentyl, 1-, 2-or 3-methylbutyl, 1-, 1, 2-or 2, 2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3-or 4-methylpentyl, 1-, 1,2-, 1,3-, 2,2-, 2, 3-or 3, 3-dimethylbutyl, 1-or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1, 2-or 1,2, 2-trimethylpropyl, more preferably for example trifluoromethyl.

It is particularly preferred that a represents an alkyl group having 1,2, 3,4,5 or 6 carbon atoms, preferably selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, trifluoromethyl, pentafluoroethyl or 1,1, 1-trifluoroethyl.

In AOne or two CH and/or CH₂The radicals may also be substituted by nitrogen, oxygen or sulfur atoms and/or by-CH = CH-groups. A therefore also represents, for example, 2-methoxyethyl.

More particularly, A represents an alkyl group having 1 to 8 carbon atoms, wherein one or two non-adjacent CH and/or CH₂The groups may be substituted by nitrogen and/or oxygen atoms, and/or wherein 1 to 7 hydrogen atoms may be substituted by F.

A' represents an alkyl group, is unbranched (linear) or branched, and contains 1,2, 3,4,5 or 6 carbon atoms. A' preferably represents methyl, and may also be ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, or may also be pentyl, 1-, 2-or 3-methylbutyl, 1-, 1, 2-or 2, 2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3-or 4-methylpentyl, 1-, 1,2-, 1,3-, 2,2-, 2, 3-or 3, 3-dimethylbutyl, 1-or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1, 2-or 1,2, 2-trimethylpropyl, more preferably for example trifluoromethyl. Preferably, a' represents an alkyl group containing 1,2, 3 or 4 carbon atoms, preferably selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or trifluoromethyl.

Ar denotes, for example, phenyl, o, m, p-tolyl, o, m, p-ethylphenyl, o, m, p-propylphenyl, o, m, p-isopropylphenyl, o, m, p-tert-butylphenyl, o, m, p-trifluoromethylphenyl, o, m, p-fluorophenyl, o, m, p-bromophenyl, o, m, p-chlorophenyl, o, m, p-hydroxyphenyl, o, m, p-methoxyphenyl, o, m, p-methylsulfonylphenyl, o, m, p-nitrophenyl, o, m, p-aminophenyl, o, m, p-methylaminophenyl, o, m, p-dimethylaminophenyl, o, m, p-aminosulfonylphenyl, o-, m-, p-methylaminosulfonylphenyl, o-, m-, p-aminocarbonylphenyl, o-, m-, p-carboxyphenyl, o-, m-, p-methoxycarbonylphenyl, o-, m-, p-ethoxycarbonylphenyl, o-, m-, p-acetylphenyl, o-, m-, p-formylphenyl, o-, m-, p-cyanophenyl, preferably also 2,3-, 2,4-, 2,5-, 2,6-, 3, 4-or 3, 5-difluorophenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3, 4-or 3, 5-dichlorophenyl, 2,4-, 2,5-, 2,6-, 3, 4-or 3, 5-dibromophenyl, 2,3,4-, 2,3,5-, 2,3,6-, 2,4, 6-or 3,4, 5-trichlorophenyl, p-iodophenyl, 4-fluoro-3-chlorophenyl, 2-fluoro-4-bromophenyl, 2, 5-difluoro-4-bromophenyl or 2, 5-dimethyl-4-chlorophenyl.

It is particularly preferred that Ar represents phenyl, which is unsubstituted or substituted by A, Hal, (CH)₂)_nHet²、(CH₂)_nOR⁵、(CH₂)_nN(R⁵)₂、(CH₂)_nCOOR⁵And/or (CH)₂)_nCON(R⁵)₂Mono-, di-or tri-substituted.

Het preferably represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyrimidinyl, triazolyl, tetrazolyl, thiadiazole, each of the above being unsubstituted or substituted by A, (CH)₂)_pHet¹、(CH₂)_pHet²OH, OA, OAr, Hal and/or (CH)₂)_pCOOR⁵Mono-, di-or tri-substituted.

Het¹Preferably represents a pyridyl group, which is unsubstituted or monosubstituted by a.

Het²Preferably represents pyrrolidinyl, piperidinyl, morpholinyl or piperazinyl, each of which is unsubstituted or substituted by OH, COOA', CON (R)⁵)₂COA and/or A monosubstitution.

R¹Preferably represents phenyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl or pyrimidinyl, each of which is unsubstitutedOr by A, OR⁵And/or CN mono-or di-substitution.

R⁵Preferably represents H, alkyl having 1,2, 3 or 4 carbon atoms, more preferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl or trifluoromethyl.

Hal preferably denotes F, Cl or Br, but may also be I, particularly preferably F or Cl.

X preferably represents CONH₂。

Y preferably represents S.

Throughout the present invention, all groups and identifiers occurring more than once may be the same or different, i.e. they are independent of each other.

The compounds of formula I may have one or more chiral centers and may therefore occur in different stereoisomeric forms. Formula I covers all these forms.

The present invention therefore relates in particular to compounds of the formula I in which at least one of the abovementioned radicals has the preferred meanings indicated above. Certain preferred groups of the compounds can be represented by the following sub-formulae Ia to Il, which correspond to formula I, and which, if no more detailed meanings are specified, have the meanings specified under formula I, but where

R in Ia¹Represents phenyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl or pyrimidinyl, each of which is unsubstituted or substituted by A, OR⁵And/or CN mono-or di-substitution;

in Ib Ar represents a phenyl group which is unsubstituted or substituted by A, Hal, (CH)₂)_nHet²、(CH₂)_nOR⁵、(CH₂)_nN(R⁵)₂、(CH₂)_nCOOR⁵And/or (CH)₂)_nCON(R⁵)₂Mono-, di-, or tri-substituted;

het in Ic represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, thiadiazole, each of which is unsubstituted or substituted by A, (CH)₂)_pHet¹、(CH₂)_pHet²OH, OA, OAr, Hal and/or (CH)₂)_pCOOR⁵Mono-, di-, or tri-substituted;

in Id Het¹Represents a pyridyl group, which is unsubstituted or monosubstituted by a;

het in Ie²Represents pyrrolidinyl, piperidinyl, morpholinyl or piperazinyl, each of which is unsubstituted or substituted by OH, COOA', CON (R)⁵)₂COA and/or A monosubstitution;

in If A represents a linear or branched alkyl group containing 1 to 8 carbon atoms, wherein one or two non-adjacent CH and/or CH₂May be substituted by nitrogen and/or oxygen atoms, and/or wherein 1 to 7 hydrogen atoms may be substituted by F;

in Ie, X represents H, CONH₂Or the CN group is selected from the group consisting of,

y represents S, and Y represents S,

r represents Ar or Het in the presence of a catalyst,

R¹represents phenyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl or pyrimidinyl, each of which is unsubstituted or substituted by A, OR⁵And/or CN is mono-or di-substituted,

ar represents a phenyl group which is unsubstituted or substituted by A, Hal, (CH)₂)_nHet²、(CH₂)_nOR⁵、(CH₂)_nN(R⁵)₂、(CH₂)_nCOOR⁵And/or (CH)₂)_nCON(R⁵)₂Mono-, di-or tri-substituted,

het represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, thiadiazole, each of which is unsubstituted or substituted by A, (CH)₂)_pHet¹、(CH₂)_pHet²OH, OA, OAr, Hal and/or (CH)₂)_pCOOR⁵Mono-, di-or tri-substituted,

Het¹represents a pyridyl group, which is unsubstituted or monosubstituted by A,

Het²represents pyrrolidinyl, piperidinyl, morpholinyl or piperazinyl, each of which is unsubstituted or substituted by OH, COOA', CON (R)⁵)₂The COA and/or the A monosubstitution,

a represents a linear or branched alkyl group containing 1 to 8 carbon atoms, wherein one or two non-adjacent CH and/or CH₂May be substituted by nitrogen and/or oxygen atoms, and/or wherein 1 to 7 hydrogen atoms may be substituted by F,

hal represents F, Cl, Br or I,

n represents 0,1, 2,3, 4or 5,

p represents 0,1 or 2,

The compounds of the general formula I and the starting materials for their synthesis are prepared, as described in the literature, by known methods (for example, standard works, such as Houben-Weyl, Methoden der organischen Chemie [ methods of organic chemistry ], Georg-Thieme-Verlag, Stuttgart) under reaction conditions which are known and suitable for the abovementioned reactions.

Variants known per se but not mentioned in more detail in the present description may also be used.

The compounds of formula I may also be prepared by reacting a compound of formula II with a compound of formula III.

The compounds of the formulae II and III are generally known. If novel, they can also be prepared by methods known per se.

Depending on the reaction conditions adopted, the reaction time is generally between a few minutes and 14 days and the reaction temperature is between-30 ℃ and 140 ℃, generally between 0 ℃ and 110 ℃, preferably between about 60 ℃ and about 110 ℃.

Examples of suitable inert solvents are: hydrocarbons, such as n-hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons, such as trichloroethylene, 1, 2-dichloroethane, tetrachloromethane, chloroform or dichloromethane; alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; ethers, such as diethyl ether, isopropyl ether, Tetrahydrofuran (THF) or dioxane; glycol ethers, such as ethylene glycol monomethyl ether or monoethyl ether or ethylene glycol dimethyl ether (diglyme); ketones such as acetone or butanone; amides, such as acetamide, dimethylacetamide or Dimethylformamide (DMF); nitriles, such as acetonitrile; sulfoxides, such as dimethyl sulfoxide (DMSO); carbon disulfide; carboxylic acids such as formic acid or acetic acid; nitro compounds, such as nitromethane or nitrobenzene; esters, such as ethyl acetate, or mixtures of the above solvents.

Particular preference is given to ethanol, toluene, ethoxyethane, acetonitrile, dichloromethane, DMF, n-methylpyrrolidone and/or water.

In the compounds of formula III, L preferably represents Cl, Br, I or a free OH group or a reactive modified OH group, for example an activated ester, imidazole (imidazole) or an alkylsulfonyloxy group containing 1 to 6 carbon atoms, preferably methylsulfonyloxy or trifluoromethylsulfonyloxy, or an arylsulfonyloxy group containing 6 to 10 carbon atoms, preferably phenyl-or p-tolylsulfonyloxy.

Generally, the reaction is carried out in the presence of an acid-binding agent, preferably an organic base such as DBU, DIPEA, triethylamine, dimethylaniline, pyridine or quinoline.

It is advantageous if an alkali or alkaline earth metal hydroxide, carbonate or bicarbonate or another salt of a weak acid of an alkali or alkaline earth metal, preferably potassium, sodium, calcium or cesium.

Depending on the reaction conditions adopted, the reaction time is generally between a few minutes and 14 days, and the reaction temperature is between-30 ℃ and 140 ℃, generally between-10 ℃ and 90 ℃, preferably between about 0 ℃ and about 70 ℃.

Examples of suitable inert solvents are hydrocarbons, such as n-hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons, such as trichloroethylene, 1, 2-dichloroethane, tetrachloromethane, trifluorotoluene, chloroform or DCM; alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; ethers, such as diethyl ether, isopropyl ether, Tetrahydrofuran (THF) or dioxane; glycol ethers such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether or ethylene glycol dimethyl ether (diglyme); ketones such as acetone or butanone; amides, such as acetamide, dimethylacetamide, N-methylpyrrolidone (NMP) or Dimethylformamide (DMF); nitriles, such as acetonitrile; sulfoxides, such as dimethyl sulfoxide (DMSO); carbon disulfide; carboxylic acids such as formic acid and acetic acid; nitro compounds, such as nitromethane or nitrobenzene; esters such as ethyl acetate, or a mixed solvent of the above solvents.

Particular preference is given to acetonitrile, dichloromethane and/or DMF.

The cleavage of the ester can be carried out using methods known to those skilled in the art.

The standard method for cleavage of esters (e.g. methyl esters) is to use boron tribromide.

Groups which can be removed by hydrogenolysis, such as the cleavage of benzyl esters, can be cleaved by reaction with hydrogen in the presence of a catalyst, for example a noble metal catalyst, such as palladium, preferably immobilized on a support such as carbon. Suitable solvents here are indicated above, in particular, for example, alcohols, such as methanol or ethanol, or amides, such as DMF. The hydrogenolysis is generally carried out at a temperature of between about 0 and 100 ℃ and a pressure of between about 1 and 200 bar, preferably between 20-30 ℃ and 1-10 bar.

The esters can be saponified, for example, using acetic acid or using NaOH or KOH in water, water/THF or water/dioxane, at a temperature between 0 and 100 ℃.

The nitrogen is alkylated under standard conditions as known to those skilled in the art.

The compounds of formula I may also be obtained by liberating the compounds of formula I from one of their functional derivatives by action with a solvolytic or hydrogenolytic agent.

Preferred starting materials for solvolysis or hydrogenolysis are compounds containing the corresponding protected amino and/or hydroxyl groups, but not one or more free amino and/or hydroxyl groups. Preference is given to those compounds which carry an amino-protecting group other than a H atom bound to the N atom, for example according to formula I, but which carry a NHR 'group (where R' denotes an amino-protecting group, such as BOC or CBZ), other than NH₂A group.

Preferred starting materials contain a hydroxyl-protecting group instead of a hydrogen atom of a hydroxyl group, for example according to formula I, but carry an R "O-phenyl group (wherein R" refers to a hydroxyl-protecting group) instead of a hydroxyphenyl group.

It is also possible for a large number of-identical or different-protected amino and/or hydroxyl groups to be present in the molecule of the starting materials. If the protecting groups present are different from one another, they can in most cases be removed by selective cleavage.

The term "amino protecting group" is a generic term referring to a group that is suitable for protecting (preventing) an amino group from chemical reaction, but which is readily removed after the chemical reaction to be performed elsewhere in the molecule is complete. These groups typically represent unsubstituted or substituted acyl, aryl, aralkoxymethyl or aralkyl groups. Since the amino-protecting groups are removed after the desired reaction (or reaction sequence), neither their type nor their size is of major importance, but preference is given to those having from 1 to 20, in particular from 1 to 8, carbon atoms. The term "acyl" is to be understood in the broadest sense relevant to the present process. It includes acyl groups derived from aliphatic, arylaliphatic, aromatic or heterocyclic carboxylic or sulfonic acids, and in particular alkoxycarbonyl, aryloxycarbonyl and aralkoxycarbonyl groups. Examples of such acyl groups are alkanoyl groups such as acetyl, propionyl and butyryl; aralkanoyl, such as phenylacetyl; aroyl groups such as benzoyl and tolyl; aralkoxyalkanoyl, such as POA; alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2, 2-trichloroethoxycarbonyl, BOC (tert-butoxycarbonyl) and 2-iodoethoxycarbonyl; aralkoxycarbonyl, such as CBZ ("benzyloxycarbonyl"), 4-methoxybenzyloxycarbonyl, and FMOC; and arylsulfonyl groups such as Mtr. Preferred amino protecting groups are BOC and Mtr, and also CBZ, Fmoc, benzyl and acetyl.

The term "hydroxy-protecting group" is also a generic term referring to a group suitable for protecting a hydroxy group from chemical reactions that can be readily removed after the chemical reactions that are to be performed elsewhere in the molecule have been completed. Typical representatives of these radicals are the abovementioned unsubstituted or substituted aryl, aralkyl or acyl radicals, and also alkyl radicals. The type and size of the hydroxyl-protecting groups are not so critical, since they are removed after the desired chemical reaction or reaction sequence, preference being given to groups having from 1 to 20, in particular from 1 to 10, carbon atoms. Examples of hydroxy protecting groups are, in particular, tert-butoxycarbonyl, benzyl, p-nitrobenzoyl, p-toluenesulfonyl, tert-butyl and acetyl, with benzyl and tert-butyl being particularly preferred. The COOH groups in aspartic and glutamic acids are preferably protected in their tert-butyl ester form (e.g., Asp (OBut)).

The compounds of the formula I are liberated from their functional derivatives-depending on the protective groups used-for example using strong acids, preferably TFA or perchloric acid, and also strong inorganic acids, such as hydrochloric acid or sulfuric acid, strong organic carboxylic acids, such as trichloroacetic acid, or sulfonic acids, such as benzenesulfonic acid or p-toluenesulfonic acid. The presence of additional inert solvents is possible, but not always necessary. Suitable inert solvents are preferably organic solvents, for example carboxylic acids such as acetic acid, ethers such as THF or dioxane, amides such as DMF, halogenated hydrocarbons such as DCM, and also alcohols such as methanol, ethanol or isopropanol, and water. Mixtures of the above solvents are also suitable. TFA is preferably used in excess, without further addition of solvent, and perchloric acid is preferably prepared by reacting acetic acid with 70% perchloric acid in a 9: the general formula of the formula 1 is used. The temperature of the cleavage reaction is preferably controlled between 0 and about 50 deg.C, preferably between 15 and 30 deg.C (room temperature).

For example, the BOC, OBut, Pbf, Pmc and Mtr groups are cleaved best using TFA in DCM or about 3-5N HCl in dioxane at 15-30 deg.C, while the FMOC group can be cleaved using about 5-50% dimethylamine, diethylamine or piperidine in DMF at 15-30 deg.C.

Protecting groups which can be removed by hydrogenolysis, such as CBZ or benzyl, can be cleaved by reaction with hydrogen in the presence of a catalyst, for example a noble metal catalyst, such as palladium, preferably immobilized on a support such as carbon. Suitable solvents here are indicated above, in particular, for example, alcohols, such as methanol or ethanol, or amides, such as DMF. The hydrogenolysis is generally carried out at a temperature of between about 0 and 100 ℃ and a pressure of between about 1 and 200 bar, preferably between 20-30 ℃ and 1-10 bar. Hydrogenolysis of the CBZ group is easily successful, for example, with 5 to 10% Pd/C in methanol, or with ammonium formate (instead of hydrogen) on Pd/C, methanol/DMF at 20-30 ℃.

Pharmaceutically acceptable salts and other forms

The compounds of the invention may be used in their final non-salt form. In another aspect, the invention also relates to the use of these compounds in the form of their pharmaceutically acceptable salts, which can be derived from various organic and inorganic acids and bases using procedures known to those skilled in the art. The pharmaceutically acceptable salt forms of the compounds of formula I are synthesized, for the most part, by conventional methods. If the compounds of the general formula I contain a carboxyl group, one of the suitable salts thereof can be prepared by reacting this compound with a suitable base to form the corresponding base addition salt. Such bases include, for example, alkali metal hydroxides including potassium hydroxide, sodium hydroxide and lithium hydroxide; alkaline earth metal hydroxides such as barium hydroxide and calcium hydroxide; alkali metal alkoxides such as sodium or potassium ethoxide and sodium or potassium propoxide; and various organic bases such as piperidine, diethanolamine and N-methylglutamine. Aluminum salts of the compounds of formula I are also included. In the case of certain compounds comprising a basic center of formula I, these compounds may be reacted with pharmaceutically acceptable organic and inorganic acids to form their acid addition salts, for example hydrogen halides, such as hydrogen chloride, hydrogen bromide or hydrogen iodide; other mineral acids and their corresponding salts, such as sulfates, nitrates, and phosphates, and the like; and alkyl-and monoaryl-sulfonic acids such as ethanesulfonic acid, p-toluenesulfonic acid and benzenesulfonic acid, as well as other organic acids and their corresponding salts such as acetate, trifluoroacetate, tartrate, maleate, succinate, citrate, benzoate, salicylate, ascorbate, and the like. Thus, pharmaceutically acceptable acid addition salts of the compounds of formula I include the following salts: acetate, adipate, alginate, arginine, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, bisulfite, bromide, butyrate, camphorate, camphorsulfonate, caprylate, chloride, chlorobenzoate, citrate, cyclopentanepropionate, gluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, fumarate, mucate (prepared from mucic acid), galacturonate, glucoheptonate, gluconate, glutamate, glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate, sodium lactobionate, malate, maleate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate, lactobionate, malate, maleate, lactate, sodium, Malonate, mandelate, metaphosphate, methanesulfonate, methylbenzoate, hydrogenphosphate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, oleate, palmoate, pectinate, persulfate, phenylacetate, 3-phenylpropionate, phosphate, phosphonate, phthalate, but this is not intended to be limiting.

Further, the basic salts of the compounds of the present invention include aluminum salts, ammonium salts, calcium salts, copper salts, iron (III) salts, iron (II) salts, lithium salts, magnesium salts, manganese (III) salts, manganous (II) salts, potassium salts, sodium salts and zinc salts, but this is not intended to be limiting. Among the above salts, ammonium salts, alkali metal salt sodium salts and potassium salts, and alkaline earth metal salt calcium salts and magnesium salts are preferably considered. Salts of the compounds of formula I with pharmaceutically acceptable non-toxic organic bases include salts of the amines primary, secondary and tertiary amines, substituted amines, as well as naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, chloroprocaine, choline, N' -dibenzylethylenediamine (benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine, meglumine, N-methyl-D-glucosamine, morpholine, piperazine, piperidine, polyamide resins, procaine, purines, theobromine, triethanolamine, procaine, and the like, Triethylamine, trimethylamine, tripropylamine, and tris (hydroxymethyl) methylamine (tromethamine), but this is not meant to be limiting.

The compounds of the present invention containing a basic nitrogen group may be used, for example, (C)₁-C₄) Agents for halogenated hydrocarbons are quaternized, such as methyl, ethyl, isopropyl and tert-butyl chlorides, bromides and iodides; two (C)₁-C₄) Alkyl sulfates such as dimethyl, diethyl, and diamyl sulfates; (C)₁₀-C₁₈) Halogenated hydrocarbons, e.g. decaneAlkyl, dodecyl, lauryl, tetradecyl and octadecyl chlorides, bromides and iodides; and aryl- (C)₁-C₄) Alkyl halides, such as benzyl chloride and phenethyl bromide. These salt compounds can be used to prepare water-soluble and oil-soluble compounds of the present invention.

The above preferred pharmaceutically acceptable salts include, but are not limited to, acetate, trifluoroacetate, benzenesulfonate, citrate, fumarate, gluconate, hemisuccinate, hippurate, hydrochloride, hydrobromide, isethionate, mandelate, meglumine, nitrate, oleate, phosphonate, pivalate, sodium phosphate, stearate, sulfate, sulfonyl salicylate, tartrate, thiomalate, tosylate and tromethamine.

Acid addition salts of the basic compounds of formula I are prepared by contacting the free base form with a sufficient amount of the desired acid in admixture to form the corresponding salt in a conventional manner. The free base is regenerable by contacting the salt form with a base and isolating the free base in a conventional manner. This free base form differs from its corresponding salt form in some sense in certain physical properties, such as solubility in polar solvents; however, for the purposes of the present invention, these salts also correspond on the other hand to their respective free base forms.

As mentioned above, pharmaceutically acceptable base addition salts of the compounds of formula I are formed by reaction with metals or amines, such as alkali and alkaline earth metals or organic amines. Preferred metals are sodium, potassium, magnesium and calcium. Preferred organic amines are N, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucosamine and procaine.

The base addition salts of the acidic compounds of the present invention are prepared by contacting the free acid form with a sufficient amount of the desired base to form the corresponding salt in a conventional manner. The free acid is regenerable by contacting the salt form with an acid and isolating the free acid in a conventional manner. This free acid form differs from its corresponding salt form in some sense in certain physical properties, such as solubility in polar solvents; however, for the purposes of the present invention, these salts also correspond on the other hand to their respective free acid forms.

If a compound of the invention contains more than one group which forms a pharmaceutically acceptable salt of this type, the invention also includes double salts. Typical double salt forms include, for example, ditartrate, diacetate, difumarate, diglucamine, diphosphate, disodium and trihydrochloride, but this is not intended to be limiting.

As can be seen from the above, the term "pharmaceutically acceptable salt" in the context of this specification refers to an active ingredient, including a compound of formula I in one of its salt forms, particularly if such salt form confers an improved pharmacokinetic profile on the active ingredient compared to its free form or any other salt form of the active ingredient used previously. The pharmaceutically acceptable salt form of the active ingredient may also provide the active ingredient for the first time with such desirable pharmacokinetic properties that it has not previously been possible and which may even have a positive effect on the pharmacodynamics of the active ingredient with respect to its in vivo therapeutic effect.

Isotope of carbon monoxide

The compounds of formula I shall also include isotopically labelled forms thereof. Isotopically-labelled forms of the compounds having the general formula I differ from the compounds only in that one or more atoms of the compound are replaced by one or more atoms having an atomic mass or mass number different from the atomic mass or mass number of the atom usually found in nature. Examples of isotopes which are readily commercially available and which can be incorporated by known methods into compounds of the formula I include hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example each²H,³H,¹³C,¹⁴C,¹⁵N,¹⁸O,¹⁷O,³¹P,³²P,³⁵S,¹⁸F and³⁶and CI. Compounds of general formula I, prodrugs thereof, or pharmaceutically acceptable salts of any of them, containing one or more of the foregoing isotopes and/or other isotopes of other atoms are understood to be part of this invention. Isotopically labelled compounds of formula I can be used in a number of advantageous ways. For example, combine with³H or¹⁴Isotopically labeled compounds of formula 1 of radioisotopes of C are useful in drug and/or substrate tissue distribution assays. The two radioisotopes, namely tritium (A), (B), (C), (³H) And carbon-14 (¹⁴C) In that respect Due to such things as deuterium (²H) The heavier isotopes of (a) have higher metabolic stability, and incorporation of such isotopically labeled compounds into compounds of formula I is therapeutically beneficial. Higher metabolic stability directly leads to an increased in vivo half-life or a reduced dose, which in most cases represents a preferred embodiment of the invention. Isotopically labeled compounds of formula I can generally be prepared by carrying out the procedures disclosed in the synthetic schemes and associated descriptions in the examples section and preparations section of this document, substituting a readily available isotopically labeled reactant for a non-isotopically labeled reactant.

To control the oxidative metabolism of compounds by first order kinetic isotope effects, deuterium (A), (B), (C), (²H) Incorporated into the compound. The first order kinetic isotope effect is due to a change in the chemical reaction rate resulting from the replacement of the isotope core due to a change in the ground state energy required to form a covalent bond after the isotope replacement. The replacement of heavier isotopes generally results in a reduction in the ground state energy of the chemical bonds, thereby causing a reduction in the rate of rate-limiting bond cleavage reactions. If bond breakage occurs in or near the saddle point region along the coordinates of the multi-product reaction, the product distribution ratio can be significantly altered. The explanation is as follows: if deuterium is bonded to a non-replaceable position of a carbon atom, the difference in rate k is usually_m/k_dAnd (2-7). If this rate difference is successfully applied to a compound of the general formula I which is susceptible to oxidation, then this rate difference isThe properties of the compound in vivo can be significantly altered, thereby improving pharmacokinetic properties.

In the discovery and development of therapeutic agents, those skilled in the art have attempted to optimize pharmacokinetic parameters while maintaining favorable in vitro properties. It is reasonable to assume that many compounds with poor pharmacokinetic properties are susceptible to oxidative metabolism. The existing in vitro liver microsome assay provides valuable information about this type of oxidative metabolic processes, which makes it possible to rationally design deuterium containing compounds of general formula I with improved stability against oxidative metabolism. Thus, the pharmacokinetic properties of the compounds of formula I are significantly improved, which can be achieved with a prolonged half-life (t/2) in vivo, the most therapeutically effective concentration (C)_max) The area under the dose response curve (AUC), and F, and also the reduced clearance, dose, and material cost.

The following description is provided to illustrate the above: compounds of formula I having multiple sites of potential oxidative metabolic attack, such as benzyl hydrogen atoms and hydrogen atoms bonded to nitrogen atoms, are prepared as a series of analogs in which various combinations of hydrogen atoms are replaced with deuterium atoms, such that some, most, or all of the hydrogen atoms are replaced with deuterium atoms. The determination of the half-life makes it possible to advantageously and accurately determine the extent to which the resistance to oxidative metabolism is improved. It was determined in this way that the half-life of the parent compound can be increased by up to 100% due to this type of deuterium-hydrogen substitution.

Deuterium-hydrogen substitution in the compounds of formula I can also be used to advantageously alter the metabolite profile of the starting compound to reduce or eliminate undesirable toxic metabolites. For example, if a toxic metabolite is produced by oxidative carbon-hydrogen (C-H) bond cleavage, it is reasonable to assume that the deuterium-containing analog will significantly reduce or eliminate the production of undesirable metabolites even if the particular oxidation reaction is not a rate-determining step. Further prior art information on deuterium-hydrogen replacement can be found, for example, in Hanzlik et al, J.org.chem.55,3992-3997,1990, Reider et al, J.org.chem.52,3326-3334,1987, Foster, adv.drug Res.14,1-40,1985, Gillette et al, Biochemistry33(10) 2927-.

The invention also relates to medicaments comprising at least one compound of the general formula I and/or pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, and optionally excipients and/or adjuvants.

The pharmaceutical preparations may be administered in the form of dosage units consisting of a predetermined amount of the active ingredient per unit dose. Depending on the condition to be treated, the mode of administration and the age, weight and physical condition of the patient, such a unit may comprise the compound of the invention in an amount of, for example, 0.5mg to 1g, preferably 1mg to 700mg, especially preferably 5mg to 100mg, or the pharmaceutical preparation may be administered in the form of a dosage unit comprising a predetermined amount of the active ingredient per unit dose. Preferred dosage unit formulations comprise a daily dose or partial dose, or a fraction thereof, of the active ingredient as described above.

In addition, pharmaceutical preparations of this type can be prepared by methods generally known to those skilled in the art of pharmacy.

The pharmaceutical formulations may be adapted for administration by any appropriate method as required, for example by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods. Such preparations may be prepared by various methods known to those skilled in the art of pharmacy, for example, by mixing the active ingredient with adjuvants or adjuvants.

Pharmaceutical formulations adapted for oral administration may be administered as separate units, for example, capsules or tablets; powder or granules; solutions or suspensions in water and non-aqueous liquids; edible foams or foam foods; or oil-in-water type liquid emulsion and water-in-oil type liquid emulsion.

Thus, for example, in the case of oral tablets or capsules, the active ingredient may be mixed with inert, oral, non-toxic, pharmaceutically acceptable excipients, for example, ethanol, glycerol, water and the like. Powders are prepared by comminuting the compound to a suitable particle size and mixing it with a pharmaceutical excipient (e.g. a food carbohydrate such as starch or mannitol) which is comminuted in a similar manner. Fragrances, preservatives, dispersants and dyes may also be present.

Capsules were prepared by first preparing a powder mixture as described above and then filling into shaped gelatin shells. Glidants and lubricants (e.g., highly dispersed silicic acid, talc, magnesium stearate, calcium stearate or polyethylene glycol in solid form) may be added to the powder mixture prior to the filling operation. Disintegrating or solubilizing agents, such as agar-agar, calcium carbonate or sodium carbonate, may also be added to improve the bioavailability of the drug after capsule administration.

In addition, if desired or necessary, suitable binders, lubricants and disintegrants and also dyes can likewise be added to the mixture. Suitable binders include starch, gelatin, natural sugars (e.g., glucose or beta-lactose, corn sweeteners), natural and synthetic gums (e.g., acacia, tragacanth) or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. The formulation of tablets is as follows, for example, by preparing a powder mixture, granulating or dry-pressing the mixture, adding a lubricant and a disintegrant, and tabletting the entire mixture to form tablets. The powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, optionally with a binder (such as carboxymethylcellulose, an alginate, gelatin or polyvinylpyrrolidone), a dissolution retardant (such as paraffin), an absorption enhancer (such as a quaternary ammonium salt) and/or an absorbent (such as bentonite, kaolin or calcium hydrogen phosphate). The powder mixture can be formed into granules by moistening with a binder such as syrup, starch paste, acadia viscose or cellulose or a solution of a polymer material and then sieving. Alternatively, the powder mixture may be run through a tableting machine to obtain non-uniform shaped pieces which are then broken up to form granules. To prevent the tablets from sticking to the tablet-pressing die, a lubricant, stearic acid, stearate, talc or mineral oil may be added to the granules. The lubricated mixture is then tableted to form tablets. The active ingredient may also be mixed with free-flowing inert excipients and then directly compressed to form tablets without the step of granulating or dry-compressing. There may also be a transparent or opaque protective layer consisting of a shellac sealing layer, a layer of sugar or polymer material, and a wax layer. Dyes may be added to these coating materials to enable differentiation between different dosage units.

Oral liquids, such as solutions, syrups and elixirs, may be prepared in the form of dosage units so that a particular quantity of the preparation contains a predetermined quantity of the compound. Syrups can be prepared by dissolving the compound in an aqueous solution with the addition of a suitable flavoring agent, while elixirs are prepared with a non-toxic alcoholic vehicle. Suspending agents are formulated so that the compound can be dispersed in a non-toxic vehicle. Solubilizing agents and emulsifiers (e.g., ethoxylated isostearyl alcohol and polyoxyethylene sorbitol ether), preservatives, flavor additives, e.g., peppermint oil or natural sweeteners or saccharin, or other artificial sweeteners, and the like, may also be added

Dosage unit formulations for oral administration may be encapsulated in microcapsules, if desired. Such formulations may also be prepared in a form in which drug release is prolonged or slowed, for example, by coating or embedding the particulate material in a polymer, wax, or like material.

The compounds of formula I and pharmaceutically acceptable salts, tautomers and stereoisomers thereof may also be administered in the form of liposome delivery systems, e.g., small unilamellar liposomes, large unilamellar liposomes and multilamellar liposomes. Liposomes can be formed from a variety of phospholipids, for example, cholesterol, stearylamine or phosphatidylcholines.

The compounds of formula I and their pharmaceutically acceptable salts, tautomers and stereoisomers may also be delivered using monoclonal antibodies as single carriers coupled to the molecules of the compounds. The compounds may also be coupled to soluble polymers as targeted drug carriers. Such polymers may include palmitoyl substituted polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenol, or polyethyleneoxidepolylysine. In addition, the compounds may be coupled to a class of biodegradable polymers suitable for achieving controlled release of the drug, such as polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates and cross-linked or amphiphilic block copolymer hydrogels.

Pharmaceutical formulations suitable for transdermal administration may be administered as discrete patches in intimate contact with the epidermis of the subject for an extended period of time. Thus, for example, the active ingredient may be delivered iontophoretically from a patch into the body, as described in general terms in Pharmaceutical Research, 3(6), 318 (1986).

The compounds suitable for topical administration may be formulated as ointments, creams, suspensions, emulsions, powders, solutions, patches, gels, sprays, aerosols or oils.

For the treatment of the eye or other external tissues, such as the mouth and skin, the best formulations for application are topical ointments or creams. In the preparation of ointments, the active ingredient may be used with a paraffinic base or a water-miscible cream base. Alternatively, the active ingredient may be formulated as a cream with an oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations suitable for topical application to the eye include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent.

Pharmaceutical formulations adapted for topical application in the mouth include lozenges, troches and mouthwashes.

Pharmaceutical preparations suitable for rectal administration may be administered in the form of suppositories or enemas.

Pharmaceutical formulations suitable for nasal administration in which the carrier material is a solid, consist of a coarse powder having a particle size, for example in the range 20 to 500 microns, and are administered by nasal inhalation, i.e. by rapid inhalation through the nasal cavity from a container of powder held in the vicinity of the nose. Suitable formulations for administration as nasal sprays or nasal drops are liquid as a carrier material, which comprises an aqueous or oily solution of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalation comprise a fine dust or aerosol of particles and may be produced by various types of pressurised dispensers, atomisers, nebulisers or insufflators.

Pharmaceutical formulations adapted for vaginal administration may be administered as pessaries, tampons, creams, gels, patches, foams or spray formulations.

Pharmaceutical formulations suitable for parenteral administration, including aqueous and non-aqueous sterile injection solutions, consisting of an antioxidant, a buffer, a bacteriostatic agent and a solute, by which means the formulation is rendered isotonic with the blood of the subject; also included are aqueous and non-aqueous sterile suspensions, which may include suspending media and thickening agents. The formulations may be administered in unit-dose or multi-dose containers, for example sealed ampoules and vials, and stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile carrier liquid, for example water for injections, immediately prior to use.

Injections and suspensions may be formulated from sterile powders, granules and tablets.

It goes without saying that these preparations may also comprise, in addition to the ingredients specifically mentioned above, other substances customary to those skilled in the art in respect of the particular type of preparation, and that, for example, preparations suitable for oral administration may contain fragrances.

The therapeutically effective amount of a compound of formula I will depend on a variety of factors including, for example, the age and weight of the animal, the precise condition to be treated and its severity, the nature of the formulation and the method of administration, and will ultimately be at the dosage determined by the attending physician or veterinarian. However, an effective dose of the compound is generally in the range of 0.1 to 100mg/kg body weight of the subject (mammal) per day, particularly usually in the range of 1 to 10mg/kg body weight per day. Thus, a typical daily actual dose for an adult mammal weighing 70kg is between 70 and 700mg, which dose may be administered daily as a single dose, or may be administered multiple times per day (e.g., two, three, four, five, six) in a series of partial doses, such that the total daily dose remains the same. An effective dose of a salt or solvate, or physiologically functional derivative thereof, may be determined as a fraction of the effective dose of the compound itself. It is envisaged that similar dosages will be suitable for the treatment of the above mentioned symptoms therein.

The invention furthermore relates to medicaments comprising at least one compound of the general formula I and/or pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, and at least one further pharmaceutically active ingredient.

The invention also relates to a kit of parts (kit) consisting of the following individual packages:

(a) an effective amount of at least one compound of the general formula I and/or pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in any ratio, and

(b) an effective amount of another pharmaceutically active ingredient.

The assembly comprises a suitable container, such as a cartridge, a separate vial, pack or ampoule. For example, the kit may comprise separate ampoules, each containing an effective amount of a compound of formula I and/or pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in any proportion, and an effective amount of a further pharmaceutically active ingredient in dissolved or lyophilized form.

Use of

The present invention relates to compounds having the general formula I for the treatment of cancer, septic shock, Primary Open Angle Glaucoma (POAG), hyperplasia, rheumatoid arthritis, psoriasis, atherosclerosis, retinopathy, osteoarthritis, endometriosis, chronic inflammation, and/or neurodegenerative diseases such as alzheimer's disease.

The present invention relates to the use of a compound having the general formula I for the preparation of a medicament for the treatment of cancer, septic shock, Primary Open Angle Glaucoma (POAG), hyperplasia, rheumatoid arthritis, psoriasis, atherosclerosis, retinopathy, osteoarthritis, endometriosis, chronic inflammation, and/or neurodegenerative diseases such as alzheimer's disease.

The present invention relates to a method of treating a mammal suffering from a disease selected from cancer, septic shock, Primary Open Angle Glaucoma (POAG), hyperplasia, rheumatoid arthritis, psoriasis, atherosclerosis, retinopathy, osteoarthritis, endometriosis, chronic inflammation, and/or neurodegenerative diseases such as alzheimer's disease, wherein the method comprises administering to the mammal a therapeutically effective amount of a compound of formula I.

The compounds of the present invention are useful as pharmaceutical active ingredients in mammals, particularly humans, for the treatment and control of cancer diseases and inflammatory diseases.

Specific cells that are sensitive to treatment with a compound of the invention can be determined in vitro. Typically, cultured cells are mixed with various concentrations of a compound of the invention for a time sufficient to allow the active agent, e.g., IgM, to induce a cellular response, e.g., expression of a surface marker, often for a period of between 1 hour and 1 week. In vitro assays can be performed using cultured cells from blood or biopsy samples. The amount of surface marker expressed is then assessed by flow cytometry using specific antibodies that recognize the marker.

In order to identify signal transduction pathways and to detect interactions between various signal transduction pathways, many scientists have developed appropriate models or model systems, such as cell culture models (e.g., Khwaja et al, EMBO, (1997), 16: 2783-93) and transgenic animal models (e.g., White et al, Oncogene, (2001), 20: 7064-. To detect certain stages of the signal transduction cascade, interacting compounds may be used to modulate the signal (e.g., Stephens et al, Biochemical J., (2000), 351: 95-105). The compounds of the invention may be used as reagents for the detection of kinase-dependent signal transduction pathway status in animal models and/or cell culture models or in clinical diseases mentioned in the present application.

Detection of kinase activity is well known to those skilled in the art. General assay systems for the detection of kinase activity are described in the literature, using substrates such as histones (e.g.Alessi et al, FEBS Lett. (1996), 399 (3): 333-.

Various test systems exist for identifying kinase inhibitors. Gamma ATP is used to detect the radioactivity of phosphorylated substrate proteins or peptides in scintillation proximity assays (Sorg et al J.of. biomolecular Screening, (2002), 7: 11-19) and in flashplate assays (flashplate assay). In the presence of the inhibitory compound, the detected radioactive signal is reduced or not detected at all. In addition, homogeneous time-resolved fluorescence resonance energy transfer (HTR-FRET) and Fluorescence Polarization (FP) techniques are suitable for use as assay methods (Sills et al, J.of Biomolecular Screening, (2002) 191-214).

Other non-radioactive ELISA assays employ specific phospho-antibodies (phospho-AB). This phospho-AB binds only to phosphorylated substrates. This binding can be detected by chemiluminescence using a peroxidase-conjugated secondary anti-sheep antibody (Ross et al, 2002, biochem. J.).

The invention covers the use of compounds of general formula I and/or physiologically acceptable salts, tautomers and solvates thereof for the preparation of a medicament for the prevention or treatment of cancer. Preferably, the cancer disease group to be treated is selected from the group consisting of brain cancer, genitourinary cancer, lymphatic cancer, stomach cancer, throat cancer, lung cancer and intestinal cancer. Another preferred group of cancer diseases are monocytic leukemia, lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma and breast carcinoma.

Also encompassed within the scope of the invention is the use of a compound of formula I and/or physiologically acceptable salts, tautomers and solvates thereof in the manufacture of a medicament for the prevention or treatment of tumor-induced diseases in a mammal in need of such treatment, in which method an effective amount of a compound of the invention is administered to the mammal in need of such treatment. The amount to be treated may vary depending on the particular disease and can be determined by one skilled in the art without undue effort.

The use for the treatment of solid tumor diseases is particularly preferred.

The solid tumor is preferably selected from the group consisting of squamous cell, bladder, stomach, kidney, head and neck, esophagus, cervix, thyroid, intestinal tract, liver, brain, prostate, genitourinary tract, lymphatic system, stomach, throat and/or lung tumors.

The solid tumor is further preferably selected from lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma, colon carcinoma and breast carcinoma.

More preferably, the compounds of the invention are used in the treatment of tumors of the blood and immune system, preferably in the treatment of tumors selected from the group consisting of acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia and/or chronic lymphocytic leukemia.

The invention also relates to the use of a compound according to the invention for the treatment of an orthopaedic disorder derived from osteosarcoma, osteoarthritis or chondropathy.

The compounds of formula I may also be administered simultaneously with other known therapeutic agents selected for their specific effectiveness against the disease to be treated.

The compounds of the present invention may also be used in combination with known anti-cancer agents. The known anticancer agents include: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenyl protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and angiogenesis inhibitors. The compounds of the invention are particularly suitable for administration simultaneously with radiotherapy.

"Estrogen receptor modulators" refers to compounds that interfere with or inhibit the binding of estrogen to the receptor by any mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4[7- (2, 2-dimethyl-1-oxopropoxy-4-methyl-2- [4- [2- (1-piperidinyl), ethoxy ] phenyl ] -2H-1-benzopyran-3-yl ] phenyl 2-1, 2-dimethylpropionate, 4,4' -dihydroxybenzophenone-2, 4-dinitrophenylhydrazine, and SH 646.

"androgen receptor modulators" refers to compounds that interfere with or inhibit the binding of androgens to the receptor by any mechanism. Examples of androgen receptor modulators include finasteride and other 5 α -reductase inhibitors, nilutamide, flutamide, bicalutamide, liazole, and abiraterone acetate.

"retinoid receptor modulators" refers to compounds that interfere with or inhibit the binding of retinoids to the receptor by any mechanism. Examples of such retinoid receptor modulators include bexarotene, tretinoin, 13-cis retinoic acid, 9-cis retinoic acid, α -difluoromethyl ornithine, ILX23-7553, trans N- (4' -hydroxyphenyl) retinoic acid, and N-4-carboxyphenylretinoic acid.

"cytotoxic agent" refers to compounds that cause cell death, or inhibit or interfere with cellular meiosis, primarily by acting directly on cellular functions, including alkylating agents, tumor necrosis factors, intercalating agents, tubulin inhibitors, and topoisomerase inhibitors.

Examples of cytotoxic agents include, but are not limited to: tirapazamine, sertenef, cachectin, ifosfamide, tasolinamine, lonidamine, carboplatin, altretamine, melphalan, prednimustine, dibromodulcitol, ramosestine, fotemustine, nedaplatin, oxaliplatin, temozolomide, heptasplatin, estramustine, improsulfan, tosylate, chloroacetocyclophosphoramide, nimustine, dibromospiro ammonium chloride, purinepedylpropane, lobaplatin, satraplatin, mitomycin methyl, cisplatin, eforufen, dexifosfamide, cis-aminodichloro (2-methylpyridine) platinum, benzylguanine, glufosfamide, GPX100, (trans ) di-mu- (hexane-1, 6-diamine) -mu- [ diamine-platinum (II) bis [ diamine (chloro) platinum (II) ] carbon tetrachloride, diamisidinylspermine, arsenic trioxide, 1- (11-dodecylamino-10-hydroxyundecyl) -3, 7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, naproxen-tebucin, valrubicin, amrubicin, antitumor ketone, 3 '-desamino-3' -morpholino-13-deoxy-10-hydroxycarminomycin, anamycin, garubicin, eletrodenide, MEN10755 and 4-demethoxy-3-desamino-3-aziridinyl-4-methanesulfonyl daunorubicin (see WO 00/50032).

Examples of tubulin inhibitors include paclitaxel, vindesine sulfate, 3 ', 4' -didehydro-4 '-deoxy-8' -nonvincalukoblastine, docetaxel, rhizomycin, dolastatin, mivobulin isethionate, auristatin, cimadrol, RPR109881, BMS184476, vinflunine, cryptophycin, 2,3,4,5, 6-pentafluoro-N- (3-fluoro-4-methoxyphenyl) benzenesulfonamide, anhydrovinblastine, N-dimethyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-tert-butylamide, TDX258 and BMS 188797.

Examples of topoisomerase inhibitors are topotecan, hycaptamine, irinotecan, rubitecan, 6-ethoxypropionyl-3 ', 4' -O-exo-benzylidene tebucin (chartreusin), 9-methoxy-N, N-dimethyl-5-nitropyrazolo [3,4,5-kl ] acridine-2- (6H) propylamine, 1-amino-9-ethyl-5-fluoro-2, 3-dihydro-9-hydroxy-4-methyl-1H, 12H-benzo [ de ] pyrano [3 ', 4': b,7] indolizino [1,2b ] quinoline-10, 13(9H,15H) dione, lurtotecan, 7- [2- (N-isopropylamino) ethyl ] - (20S) camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzosin, 2 '-dimethylamino-2' -deoxyetoposide, GL331, N- [2- (dimethylamino) ethyl ] -9-hydroxy-5, 6-dimethyl-6H-pyrido [4,3-b ] carbazole-1-carboxamide, asulamine, (5a,5aB,8aa,9b) -9- [2- [ N- [2- (dimethylamino) ethyl ] -N-methylamino ] ethyl ] -5- [ 4-hydroxy-3, 5-dimethoxyphenyl ] -5,5a,6,8,8a, 9-hexahydrofuro (3 ', 4': 6, 7) naphtho (2,3-d) -1, 3-dioxol-6-one, 2,3- (methylenedioxy) -5-methyl-7-hydroxy-8-methoxybenzo [ c ] phenanthridine, 6, 9-bis [ (2-aminoethyl) amino ] benzo [ g ] isoquinoline-5, 10-dione, 5- (3-aminopropylamino) -7, 10-dihydroxy-2- (2-hydroxyethylaminomethyl) -6H-pyrazolo [4,5,1-de ] acridin-6-one, N- [1- [2 (diethylamino) ethylamino ] -7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl ] carboxamide, N- (2- (dimethylamino) ethyl) acridine-4-carboxamide, 6- [ [2- (dimethylamino) ethyl ] amino ] -3-hydroxy-7H-indeno [2,1-c ] quinolin-7-one, and dimesna.

"antiproliferative agents" include antisense RNA and DNA oligonucleotides, such as G3139, ODN698, RVASKRAS, GEM231 and INX3001, and antimetabolites, such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, galocidine, cytarabine octadecyl phosphate, fosetabine sodium hydrate, raltitrexed, palettridid, ethimidifluoride, thiazoluraline, decitabine, nolatrexed, pemetrexed, nelzabine, 2 ' -deoxy-2 ' -methylenecytidine, 2 ' -fluoromethylene-2 ' -deoxycytidine, N- [5- (2, 3-dihydrobenzofuranyl) -sulfonyl ] -N ' - (3, 4-dichlorophenyl) urea, N6- [ 4-deoxy-4- [ N2- [2(E),4(E) -tetradecameridyl ] dienyl ] -L-glycero-B-glycinamide-B- L-mannohept-pyranosyl ] adenine, aplidine, ecteinascidin, troxacitabine, 4- [ 2-amino-4-oxo-4, 6,7, 8-tetrahydro-3H-pyrimido [5,4-b ] -1, 4-thiazin-6-yl- (S) -ethyl ] -2, 5-thenoyl-L-glutamic acid, aminopterin, 5-fluorouracil, alafenacin, 11-acetyl-8- (carbamoyloxymethyl) -4-formyl-6-methoxy-14-oxa-1, 11-diazepicyclo (7.4.1.0.0) tetradec-2, 4, 6-trien-9-ylacetate, octahydroindolizinetriol, dihydroindolizine, Lometrexol, dexrazoxane, methioninase, 2 '-cyano-2' -deoxy-N4-palmitoyl-1-B-D-arabinofuranosyl cytosine, and 3-aminopyridine-2-carbaldehyde phenylthiourea. "antiproliferative agents" also include growth factor monoclonal antibodies such as trastuzumab and tumor suppressor genes such as p53, in addition to those listed under "angiogenesis inhibitors", which can be delivered via recombinant virus-mediated gene transfer (see, e.g., US patent No. 6,069,134).

Inhibition assay for IKK epsilon

IKK epsilon-kinase assay (IKKepsilon)

SUMMARY

Kinase assays are performed in 384-well Flashplate assays (e.g., Topcount measurements).

A total volume of 50. mu.l (10mM MOPS, 10mM magnesium acetate, 0.1mM EGTA,1mM dithiothreitol, 0.02% Brij35,0.1% BSA,0.1% BioStab, pH7.5) of 1nM IKK ε, 800nM biotinylated I B (19-42) peptide (biotin-C6-C6-GLKKERLLDDRHDSGLDSMKDEE) and 10. mu.M ATP (with 0.3. mu. Ci of added) with or without test compound was added³³P-ATP/well) was incubated at 30 ℃ for 2 hours. The reaction was stopped with 25. mu.l of 200mM EDTA. After 30 minutes at room temperature, the liquid was removed and each well was washed three times with 100 μ l of 0.9% sodium chloride solution. Nonspecific responses were determined with 3. mu. MMSC2119074 (BX-795). Radioactivity was measured by Topcount (PerkinElmer). Computing results (e.g., IC) with programmatic tools provided by the IT department (e.g., AssayExplorer, Symyx)₅₀-value).

Inhibition test of TBK1

Enzyme assay

SUMMARY

Kinase assay assays were performed in 384-well scintillation plate assays (e.g., Topcount measurements).

A total volume of 50. mu.l (10mM MOPS, 10mM magnesium acetate, 0.1mM EGTA,1mM DTT,0.02% Brij35,0.1% BSA, pH7.5) of 0.6nM TANK-binding kinase (TBK1), 800nM biotinylated MELK-derived peptide (biotin-Ah-Ah-AKPKGNKDYHLQTCCGSLAYRRR) and 10. mu.M ATP (with 0.25. mu. Ci added) with or without test compound was added³³P-ATP/well) was incubated at 30 ℃ for 120 minutes. The reaction was stopped with 25. mu.l of 200mM EDTA. After 30 minutes at room temperature, the liquid was removed and each well was washed three times with 100 μ l of 0.9% sodium chloride solution. Radioactivity was measured by Topcount (PerkinElmer). The results (e.g., IC 50-values) were calculated with procedural tools (e.g., AssayExplorer, Symyx) provided by the IT department.

Cell assay

Dose-responsive phosphorus-IRF 3 inhibits Ser386

cell/MDAMB 468/INH/PHOS/IMAG/pIRF3

1. Range of

Although TBK1 and IKK epsilon are known to be key players of the innate immune response, recent findings have shown that TBK1 and IKK epsilon also play a role in Ras-induced oncogenic transformation. TBK1 was identified as the RalB effector in the Ras-like (RAL) -guanine nucleotide exchange factor (GEF) pathway required for Ras-induced transformation. TBK1 directly activates IRF3, which upon phosphorylation homodimerizes and translocates to the nucleus, and in the nucleus activates processes involved in inflammation, immune regulation, cell survival and proliferation.

The objective of this assay was to assess the efficacy of TBK1/IKK epsilon inhibitor compounds based on immunocytochemical detection of nuclear localization phosphorus-IRF 3, which is a target directly downstream of TBK 1.

Treatment with poly inosine-polycytidylic nucleotide (poly (I: C)) which is a synthetic analogue of double stranded RNA (dsrna) that is a molecular pattern associated with viral infection that is recognized by the Toll-like receptor 3(TLR3) induces TBK1/IKK epsilon activity and phosphorylates IRF3 at Ser 386.

2. Overview of the experiment

The first day: MDA-MB-468 cells were eluted with HyQ-Tase, counted, and a total volume of 35. mu.l of complete medium was seeded into transparent bottom TC-surface 384-well plates at a density of 10,000 cells per well. Alternatively, cells in the cryovial can also be planted directly.

The next day: cells were pretreated with inhibitor compounds for 1 hour prior to stimulation with Poly (I: C). After 2 hours incubation with Poly (I: C), the cells were fixed in Paraformaldehyde (PFA) and permeabilized with methanol (MeOH). The cells were then blocked and incubated with anti-pIRF 3 antibody overnight at 4 ℃.

And on the third day: the primary antibody was washed away, a secondary antibody conjugated to AlexaFluor88 was added, the cells were counterstained with propidium iodide, and images were taken in an IMX Ultra high content reader.

3. Reagents, materials

Cells: ATCC HTB132, Burger's laboratory (MP-CB2010-327 or MDA-MB-468/10)

Plate medium = medium：

RPMI1640,Invitrogen#31870

10%FCS,Invitrogen#10270-106

2mM Glutamax,Invitrogen#35050-038

1mM sodium pyruvate, Invitrogen #11360

1%Pen/Strep

37℃,5%CO₂

Flat plate: black/clear bottom 384 well cell culture plates, Falcon #353962 or Greiner #781090

Transferring seeds：HyQ-Tase,Thermo Scientific(HyClone)#SV30030.01

Other reagents：

Poly (I: C) (LMW), Invivogen # tlrl-picw (20 mg/ml stock prepared with sterile PBS, denatured in a55 ℃ water bath for 30 minutes, slowly cooled to room temperature, and stored in aliquots at-20 ℃.

Reference inhibitor MSC2119074A-4= BX-795(IC50:200-800nM)

Inhibition control 10 μ M MSC2119074A-4= BX-795

Neutral control 0.5% DMSO

Each experiment included a 10-point dose response curve with MSC2119074A-4= BX-795.

Hepes,Merck#1.10110

PBS1x DPBS,Invitrogen#14190

Formaldehyde (methanol-free, 16%, ultra pure EM grade), Polysciences #18814 (stored at room temperature), final concentration: 4%

Methanol, Merck #1.06009.1011(-20 ℃ Pre-Cooling)

Goat serum, PAA # B15-035 (stored at 4 deg.C for a long period at 20 deg.C), final concentration: 10%

BSA (no IgG and protease, 30%), US-Biological # A1317 (stored at 4 ℃ C., long-term storage at 20 ℃ C.), final concentration: 2%

Tween 20 detergent, Calbiochem #655204 (storage at room temperature), (10% stock solution prepared with water; final concentration: 0.1%)

anti-pIRF-3 rabbit mAb, Epitomics #2526-B (stored at-20 ℃ C.), final concentration 1:2000 in PBS/2% BSA

Alexa Fluor goat-anti-rabbit-488, Invitrogen # A11034or # A11008 (stored at 4 ℃ C., protected from light), final concentration: 1:2000 in PBS/2% BSA/0.1% Tween

Propidium Iodide (PI), Fluka #81845,1mg/ml in water (stored at 4 ℃ C., protected from light) to a final concentration of 0.2. mu.g/ml

4. Step (ii) of

HPLC/MS conditions:

chromatographic column Chromolith speedROD RP-18e,50x4.6mm²

Gradient A: B =96:4 to 0:100

Flow rate of 2.4ml/min

Eluent A is water and 0.05 percent formic acid

Eluent B is acetonitrile +0.04% formic acid

Wavelength of 220nm

Mass spectrometry Positive ion mode

¹H NMR coupling constant J [ Hz ]]。

Example 1

Preparation of 5- (3, 4-dimethoxy-benzoylamino) -2-phenyl-thiazole-4-carboxamide ("A1")

1.1 N²-benzoyl-3-nitrilopropionamide

To a solution of 2-amino-2-cyanoacetamide (8g, 0.08mol) in pyridine (10mL) was added a solution of benzoyl chloride (0.08mol) in pyridine (15mL), and the mixture was stirred at room temperature for 40 minutes. Concentration under reduced pressure then gave the crude product which was used directly in the next step without further purification.

1.2 N²-benzoyl-3-amino-3-thioacrylamide

To a crude solution of 2-acylamino-2-cyanoacetamide (1g) in ethanol (30mL) and methanol (10mL) was added triethylamine (0.69 mL). Hydrogen sulfide was passed through the solution at 40-42 ℃ for 6 hours. The precipitated solid was collected by filtration. The filtrate was again sparged with hydrogen sulfide at 40 ℃ until saturation, held overnight, and the resulting solid collected. The solids were combined and recrystallized from ethanol to give the title compound in 50% yield over two steps:¹H NMR(400MHz，DMSO-d₆)δ[ppm]5.3(d，J=8.2Hz，1H)；7.5(s，1H)；7.5(t，J=7.3Hz，2H)；7.6(m，1H)；7.6(s，1H)；7.9(m，2H)；8.2(d，J=8.0Hz，1H)；9.4(s，1H)；10.0(s，1H)。

similarly, the following compounds were obtained:

N²- (3-pyridinecarbonyl) -3-amino-3-thioacrylamide:¹H NMR(400MHz，DMSO-d₆)δ[ppm]5.4(d，J=8.1Hz，1H)；7.5(s，1H)；7.5(dd，J=7.8，4.9Hz，1H)；7.7(s，1H)；8.2(d，J=8.1Hz，1H)；8.5(d，J=8.1Hz，1H)；8.7(d，J=3.4Hz，1H)；9.0(d，J=1.7Hz，1H)；9.4(s，1H)；10.0(s，1H)。

1.35-amino-2-phenyl-1, 3-thiazole-4-carboxamides

Will N²A mixture of (1g) of-benzoyl-3-amino-3-thioacrylamide and (15g) of polyphosphoric acid was heated at 140 ℃ for 8 hours with stirring. The resulting yellow-orange solution was cooled to room temperature, quenched with water (50mL), and the pH of the quenched solution was adjusted to 6-7 with 50% aqueous potassium hydroxide. The resulting precipitate was collected by filtration to give the title compound in 75% yield:¹H NMR(400MHz，DMSO-d₆)δ[ppm]7.1(s，1H)；7.3(s，1H)；7.4(m，3H)；7.4(t，J=7.6Hz，2H)；7.8(d，J=8.0Hz，2H)。

similarly, the following compounds were obtained:

5-amino-2-pyridin-3-yl-1, 3-thiazole-4-carboxamide:¹H NMR(400MHz，DMSO-d₆)δ[ppm]7.1(s，1H)；7.4(s，1H)；7.4(m，3H)；8.1(m，1H)；8.5(dd，J=4.8,1.3Hz，1H)；9.0(d，J=2.0Hz，1H)，

5-amino-2-pyridin-4-yl-1, 3-thiazole-4-carboxamide:¹H NMR(400MHz，DMSO-d₆)δ[ppm]7.2(s，1H)；7.4(s，1H)；7.6(wide s，2H)；7.8(d，J=4.84Hz，2H)；8.6(d，J=4.84Hz，2H)。

45- [ (3, 4-Dimethoxybenzoyl) amino ] -2-phenyl-1, 3-thiazole-4-carboxamide ("A1"):

to a solution of 5-amino-2-phenyl-1, 3-thiazole-4-carboxamide (0.3g, 1.4mmol) in NMP (1mL) was added a solution of 3, 4-dimethoxybenzoyl chloride (0.41g, 2.0mmol) in NMP (3mL) and DBU (0.21g, 1.4mmol) was added and the mixture was stirred at reflux for 4 h. The resulting precipitate was collected by filtration and washed with NMP, acetonitrile, ethanol, ether to give 0.1g (30%) of the title compound. The compound was purified by reverse phase HPLCAnd (3) conversion:¹H NMR(400MHz，DMSO-d₆)δ[ppm]3.9(s，6H)；7.2(d，J=8.3Hz，1H)；7.5(m，5H)；8.0(m，3H)；8.1(s，1H)；12.7(s，1H)。

similarly, the following compounds were obtained:

5- [ (3, 4-dimethoxybenzoyl) amino ] -2-pyridin-3-yl-1, 3-thiazole-4-carboxamide ("a 2"):

¹H NMR(400MHz，DMSO-d₆)δ[ppm]3.9(s，7H)7.2(d，J=8.1Hz，1H)7.5(s，1H)7.5(d，J=7.3Hz，1H)7.7(m，1H)8.0(s，1H)8.2(s，1H)8.5(d，J=7.6Hz，1H)8.7(d，J=4.6Hz，1H)9.3(s，1H)12.7(s，1H)。

example 2

Preparation of 4- [4- (4-carbamoyl-2-pyridin-4-yl-thiazol-5-ylcarbamoyl) -benzyl ] -piperazine-1-carboxylic acid tert-butyl ester ("A3")

A mixture of 4- ((4-tert-butoxycarbonyl) piperazin-1-yl) methyl) benzoic acid (300mg, 0.936mmol, 1.0 eq), 5-amino-2- (pyridin-4-yl) thiazole-4-carboxamide (206, 0.936,1.0 eq), HATU (356mg, 0.936mmol, 1.0 eq) and N-methylmorpholine (106. mu.l, 0.936mmol, 1.0 eq) in DMF (8ml) was heated at 75 ℃ for 30 min. DBU (285. mu.l, 1.873mmol, 2.0 equiv.) was added and the mixture was heated at 90 ℃ for 4 h.

The solvent was evaporated in vacuo and the residue was redissolved in water (20ml), extracted with ethyl acetate (20ml × 3 times), the organic phase washed with brine, Na₂SO₄Drying, filtering and evaporating. The residue was triturated in methanol, filtered and dried to give the title compound as an off-white powder.

HPLC method: a-aqueous 0.1% TFA, B-acetonitrile 0.1% TFA: flow rate-2.0 ml/min.

Column: x Bridge C8(50 x4.6mm.3.5. mu.).

The following compounds were obtained with reference to the above examples:

example 3

Preparation of 2- (2, 6-dimethyl-pyridin-4-yl) -5- [4- (4-methyl-piperazin-1-ylmethyl) -benzoylamino ] -thiazole-4-carboxamide ("A16

With reference to the following flow chart,

3.1N- (2-amino-1-cyano-2-oxoethyl) -2, 6-dimethylisonicotinamide

Stirring 2-amino 2-cyanoacetamide (1.4g, 0.01473mol, 1 equivalent) in pyridine (15ml) at room temperature for 1 hour, adding 2, 6-dimethylisonicotinoyl chloride (2.5g, 0.1473mol, 1 equivalent) thereto at 0 deg.C [ prepared by stirring 2, 6-dimethylisonicotinic acid (2.5g, 0.0465mol, 1 equivalent) in thionyl chloride (15ml) at 85 deg.C for 2 hours, and removing thionyl chloride under nitrogen atmosphere and vacuum ], the reaction mixture was stirred at room temperature for 14 hours, after completion of the reaction, pyridine was removed in vacuum, and the crude product was purified by column chromatography to give the title compound;

yield: 39 percent; MS (ESI +): 233.05.

3.2N- (1, 3-diamino-1-oxo-3-thiopropan-2-yl) -2, 6-dimethylisonicotinamide

N- (2-amino-1-cyano-2-oxoethyl) -2, 6-dimethylisonicotinamide (1.5g, 0.00646mol, 1 equiv.), sodium hydrosulfide (1.05g, 0.00646mol, 3 equiv.) in water/1, 4-dioxane, diethylamine hydrochloride (1.0 g. after completion of the reaction, the reaction was cooled, water was added, extraction was performed with ethyl acetate, drying over sodium sulfate, evaporation of the solvent, recrystallization of the crude product in diethyl ether gave 1.0g N- (1, 3-diamino-1-oxo-3-thiopropan-2-yl) -2, 6-dimethylisonicotinamide, yield: 64%; MS (ESI +): 267;

¹H NMR400MHz，DMSO-d₆：δ[ppm]9.98(s，1H)，9.40(s，1H)，8.39(d，J=8.00Hz，1H)，7.64(s，1H)，7.48(s，1H)，7.45(d，J=8.00Hz，1H)，3.55(s，6H)。

3.35-amino-2- (2, 6-dimethylpyridin-4-yl) thiazole-4-carboxamide

N- (1, 3-diamino-1-oxo-3-thiopropan-2-yl) -2, 6-dimethylisonicotinamide (1.0g, 0.00375mol) was treated with polyphosphoric acid (4g) and heated to 140 ℃ for 1 hour. The resulting yellow-orange solution was cooled to room temperature, quenched with water, adjusted to pH 6-7 with 50% aqueous potassium hydroxide, extracted with ethyl acetate, dried over sodium sulfate and evaporated. The crude product was recrystallized from diethyl ether to give 0.200mg 5-amino-2- (2, 6-dimethylpyridin-4-yl) thiazole-4-carboxamide; yield: 21.5 percent; MS (ESI +): 248.

3.42- (2, 6-dimethyl-pyridin-4-yl) -5- [4- (4-methyl-piperazin-1-ylmethyl) -benzoylamino ] -thiazole-4-carboxylic acid amide

4- (4-methylpiperazine-methyl) benzoic acid (0.266g, 0.001209mol) and Carbonyldiimidazole (CDI) (0.293g0.00181mole) were dissolved in dry DMF (5ml), stirred at 90 ℃ for 1 hour, then 5-amino-2- (2, 6-dimethylpyridin-4-yl) thiazole-4-carboxamide (0.15g, 0.00064mol) was added and the mixture stirred at 90 ℃ for 14 hours. Completion of the reaction was observed by TLC, the solvent was evaporated and the crude product was purified by column chromatography (basic alumina) to give 17.6mg of the title compound; yield: 9 percent; MS (ESI +): 465.00, respectively;

¹H NMR400MHz，DMSO-d₆：δ[ppm]12.77(s，1H)，8.21(s，1H)，8.05(s，1H)，7.91(d，J=8.00Hz，2H)，7.67(s，2H)，7.56(d，J=7.76Hz，2H)，3.55(s，2H)，2.48(s，6H)，2.38-2.40(m，8H)，2.17(s，1H)；

HPLC >98%, residence time (min): 1.969.

example 4

Preparation of 5- [4- (4-methyl-piperazin-1-ylmethyl) -benzoylamino ] -2- (3-trifluoromethyl-pyridin-4-yl) -thiazole-4-carboxamide ("A17

"A17" was synthesized according to the procedure described in the preparation method of "A16" (steps 1 to 4).

4.1N- (2-amino-1-cyano-2-oxoethyl) -3- (trifluoromethyl) isonicotinamide

The title compound was synthesized according to the procedure described in step 1 of the preparation method of "a 16"; yield: 18.5 percent; MS (ESI +): 273.05, respectively;

¹H NMR400MHz，DMSO-d₆：δ[ppm]9.97(d，J=8.00Hz，1H)，9.05(s，1H)，0.00(d，J=4.00Hz，1H)，7.90(s，1H)，7.79(t，J=4.92Hz，2H)，5.70(d，J=8.00Hz，1H)。

4.2N- (1, 3-diamino-1-oxo-3-thiopropan-2-yl) -3- (trifluoromethyl) isonicotinamide:

the title compound was synthesized according to the procedure described in step 2 of the preparation method of "a 16"; yield: 55 percent; MS (ESI +): 304.1.

4.35-amino-2- (3- (trifluoromethyl) pyridin-4-yl) thiazole-4-carboxamide:

the title compound was synthesized according to the procedure described in step 3 of the preparation of "a16", yield: 18 percent; MS (ESI +): 289.0.

4.45- [4- (4-methyl-piperazin-1-ylmethyl) -benzoylamino ] -2- (3-trifluoromethyl-pyridin-4-yl) -thiazole-4-carboxamide ("A17")

The title compound was synthesized according to the procedure described in step 4 of the preparation method of "a 16"; yield: 20 percent; MS (ESI +): 505.30, respectively;

¹H NMR400MHz，DMSO-d₆：δ[ppm]12.75(s，1H)，9.12(s，1H)，9.00(s，1H)，8.10-8.00(m，1H)，7.97(d，J=4.00Hz，2H)，7.92(d，J=8.00Hz，1H)，7.83-7.85(m，1H)，7.55-7.50(m，2H)，3.56(s，2H)，2.39-2.45(m，8H)，2.20(s，1H)；

HPLC > 98%; residence time (min): 2.911.

with reference to example "a16", the following compounds were obtained:

the following compounds were obtained with reference to the above examples:

example 5

5- (4-methylbenzamido) -2-pyridin-4-yl-thiazole-4-carboxylic acid amide ("A55") was prepared according to the following scheme,

4-Methylbenzoic acid (0.054g, 0.0004mol) and Carbonyldiimidazole (CDI) (0.097g, 0.0006mol) were dissolved in dry DMF (2ml), stirred at 90 ℃ for 1 hour, then 5-amino-2-pyridin-4-yl-thiazole-4-carboxamide (0.05g, 0.0002mol) was added and the mixture was stirred at 90 ℃ overnight. The reaction was complete as observed by TLC, the solvent was evaporated and the crude product was purified by silica gel column chromatography to give 17.6mg5- (4-methyl-benzoylamino) -2-pyridin-4-yl-thiazole-4-carboxamide ("a55") (17.6mg, 32.46% yield);

LCMS: measured value (M)⁺，339.0)

HPLC method: a-aqueous solution of 0.1% TFA, B-: 0.1% TFA in acetonitrile, flow-1.0 ml/min.

Column: xbridge C8(50X4.6mm, 3.5. mu)

Residence time (min): 3.18 of;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.79(s，1H)，8.71(d，J=4.00Hz，2H)，8.23(s，1H)，8.06(s，1H)，7.99-7.97(m，2H)，7.86(d，J=8.00Hz，2H)，7.46(d，J=8.00Hz，2H)，2.42(s，3H)。

similarly, the following compounds were obtained.

5-Benzoylamino-2-pyridin-4-yl-thiazole-4-carboxamide ("A56")

LCMS: measured value (M)⁺325.0); HPLC: residence time (min): 2.83;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.83(s，1H)，8.72-8.71(m，2H)，8.25(s，1H)，8.08(s，1H)，8.00-7.96(m，4H)，7.73(t，J=16.00Hz，1H)，7.68-7.64(m，2H)；

5- { [4- (dimethylamino) benzoyl ] amino } -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A57")

LC-MS: found (M +, 368.0); HPLC: residence time (min): 3.10;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.59(s，1H)，8.69(d，J=6.08Hz，2H)，8.16(s，1H)，8.00(s，1H)，7.96(d，J=6.12Hz，2H)，7.77(d，J=9.08Hz，2H)，6.85(d，J=9.12Hz，2H)，3.04(s，6H)；

5- [ (4-aminobenzoyl) amino ] -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A58")

LC-MS: found (M +, 340.0); HPLC: residence time (min): 2.061, respectively;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.54(s，1H)，8.74(s，2H)，8.19(s，1H)，8.07(d，J=5.76Hz，2H)，8.00(s，1H)，7.65(d，J=8.72Hz，2H)，6.68(d，J=8.72Hz，2H)；

5- { [ (1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A59")

LC-MS: found (M +, 328.0); HPLC: residence time (min): 2.71;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.38(s，1H)，8.69(d，J=6.12Hz，2H)，8.14(s，1H)，δ7.99-7.95(m，1H)，7.22-7.21(m，1H)，6.87-6.85(m，1H)，6.25-6.23(m，1H)，3.95(s，3H)；

5- (2-furoylamino) -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A60")

LC-MS: found (M +, 315.0); HPLC: residence time (min): 2.26;

NMR Analysis

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.54(s，1H)，8.70(d，J=6.12Hz，2H)，8.21(brs，1H)，8.11-8.11(m，1H)，7.98(d，J=6.16Hz，2H)，7.45(d，J=4.28Hz，1H)，6.82(d，J=5.32Hz，1H)；

2-pyridin-4-yl-5- [ (1H-pyrrol-2-ylcarbonyl) amino ] -1, 3-thiazole-4-carboxamide ("A61")

LC-MS: found (M +, 314.0); HPLC: residence time (min): 2.24;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.33(s，1H)，12.20(s，1H)，8.69(d，J=6.16Hz，2H)，8.15(brs，1H)，7.96-7.95(m，2H)，7.99(brs，1H)，7.17-7.15(m，1H)，6.83-6.81(m，1H)，6.31-6.29(m，1H)；

2-pyridin-4-yl-5- [ (2-thienylcarbonyl) amino ] -1, 3-thiazole-4-carboxamide ("A62")

LC-MS: found (M +, 331.0); HPLC: residence time (min): 2.522, respectively;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.69(s，1H)，8.71(d，J=5.96Hz，2H)，8.24(brs，1H)，8.06(d，J=5.08Hz，2H)，7.97(d，J=5.92Hz，2H)，7.82-7.81(m，1H)，7.33(t，J=4.48Hz，1H)；

5- [ (1H-pyrazol-3-ylcarbonyl) amino ] -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A63")

LC-MS: found (M +, 315.0); HPLC: residence time (min): 2.05;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]13.72(s，1H)，12.63(s，1H)，8.70(d，J=6.12Hz，2H)，8.11(brs，1H)，8.01(brs，1H)，7.98-7.97(m，2H)，7.94-7.93(m，1H)，6.90-6.89(m，1H)；

5- (3-furoylamino) -2-pyridin-4-yl-1, 3-thiazole-4-carboxamide ("A64")

LC-MS: found (M +, 315.0); HPLC: residence time (min): 2.28;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.35(s，1H)，8.70(d，J=6.08Hz，2H)，8.54(s，1H)，8.20(brs，1H)，8.26(brs，1H)，7.97(d，J=6.16Hz，2H)，7.94-7.94(m，1H)，6.85-6.85(m，1H)；

2-pyridin-4-yl-5- [ (3-thienylcarbonyl) amino ] -1, 3-thiazole-4-carboxamide ("A65")

LC-MS: found (M +, 331.0); HPLC: residence time (min): 2.66;

¹H NMR(400MHz，DMSO-d₆)δ[ppm]12.56(s，1H)，8.71(d，J=6.12Hz，2H)，8.41(d，J=4.24Hz，1H)，8.22(s，1H)，8.06(s，1H)，7.98(d，J=6.12Hz，2H)，7.81(d，J=7.96Hz，1H)，7.53(d，J=6.44Hz，1H)。

compounds according to the invention inhibit IC of TBK1 and IKK epsilon₅₀Value of

The following examples relate to medicaments:

example A: injection bottle

A solution of 100g of the compound of the formula I as active ingredient and 5g of sodium dihydrogenphosphate in 3L of redistilled water is adjusted to pH 6.5 with 2N hydrochloric acid, sterile-filtered, transferred into injection vials, freeze-dried under sterile conditions and sealed under sterile conditions. Each solution bottle contained 5mg of active ingredient.

Example B: suppository

20g of the compound of the formula I as active ingredient are mixed with 100g of soya lecithin and 1400g of cocoa butter, poured into moulds and cooled. Each suppository contains 20mg of active ingredient.

Example C: solution formulation

1g of compound of general formula I as active ingredient, 9.38g of NaH₂PO₄·2H₂O、28.48gNa₂HPO₄·12H₂O and 0.1g benzalkonium chloride in 940mL redistilled water. The pH of the solution was adjusted to 6.8, the solution was made up to 1L and sterilized by radiation. The solution is used in the form of an eye drop.

Example D: ointment

500mg of a compound of the general formula I as active ingredient are mixed with 99.5g of vaseline under sterile conditions.

Example E: tablet formulation

1kg of a compound of the general formula I as active ingredient, 4kg of lactose, 1.2kg of potato flour, 0.2kg of talc and 0.1kg of magnesium stearate are compressed into tablets in accordance with the customary method so that each tablet contains 10mg of active ingredient.

Example F: coated tablet

Tablets were compressed analogously to example E and then coated in a conventional manner with a sucrose coating, potato starch, talc, tragacanth and dye.

Example G: capsule preparation

2kg of a compound of the formula I as active ingredient are introduced into hard capsules in a conventional manner so that each capsule contains 20mg of active ingredient.

Example H: ampoule agent

A solution of 1kg of the compound of the formula I as active ingredient in 60L of redistilled water is sterile-filtered, transferred into ampoules, lyophilised under sterile conditions and sealed under sterile conditions. Each ampoule contains 10mg of active ingredient.

Claims

1. A compound of the general formula I,

in the formula (I), the compound is shown in the specification,

y represents NH, N-Me, S or O,

r represents Ar or Het in the presence of a catalyst,

R¹represents a phenyl group, a furyl group,Thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl, pyrimidinyl, pyridazinyl, indolyl, isoindolyl, benzimidazolyl, indazolyl, quinolinyl, 1, 3-benzodioxolyl, benzothienyl, benzofuranyl, imidazopyridinyl, or furo [3,2-b ] group]Pyridyl, each of the above radicals being unsubstituted OR substituted by Hal, A, OR⁵、CN、COOA、COOH、CON(R⁵)₂And/or NR⁵The COA' is mono-or di-substituted,

Het²represents a dihydropyrrolyl group, a pyrrolidinyl group, a tetrahydroimidazolyl group, a dihydropyrazolyl group, a tetrahydropyrazolyl group, a dihydropyridinyl group, a tetrahydropyridinyl group, a piperidyl group, a morpholinyl group, a hexahydropyridazinyl group, a hexahydropyrimidyl group, [1, 3]]Dioxolanyl, piperazinyl, each of which is unsubstituted or substituted by OH, COOA', CON (R)⁵)₂The COA and/or the A monosubstitution,

hal represents F, Cl, Br or I,

n represents 0,1, 2,3, 4or 5,

p represents 0,1 or 2,

q represents 1,2, 3 or 4,

2. The compound of claim 1, wherein

R¹Represents phenyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl or pyrimidylEach of said radicals being unsubstituted or substituted by A, OR⁵And/or CN is mono-or di-substituted,

3. A compound according to claim 1 or 2, wherein

Ar represents phenyl which is unsubstituted or substituted by A, Hal, (CH)₂)_nHet²、(CH₂)_nOR⁵、(CH₂)_nN(R⁵)₂、(CH₂)_nCOOR⁵And/or (CH)₂)_nCON(R⁵)₂Mono-, di-or tri-substituted,

4. A compound as claimed in one or more of claims 1 to 3, wherein

5. The compound as claimed in one or more of claims 1 to 4, wherein

Het¹Represents pyridyl, which is unsubstituted or monosubstituted by A,

6. The compound as claimed in one or more of claims 1 to 5, wherein

7. The compound as claimed in one or more of claims 1 to 6, wherein

A represents a linear or branched alkyl group containing 1 to 8 carbon atoms, wherein one or two non-adjacent CH and/or CH₂The radicals being substituted by nitrogen and/or oxygen atoms, and/or wherein 1 to 7 hydrogen atoms may be substituted by F,

8. The compound as claimed in one or more of claims 1 to 7, wherein

r represents Ar or Het in the presence of a catalyst,

Het¹represents pyridyl, which is unsubstituted or monosubstituted by A,

hal represents F, Cl, Br or I,

n represents 0,1, 2,3, 4or 5,

p represents 0,1 or 2,

9. The compound of claim 1, selected from the group consisting of,

10. A process for the preparation of compounds of the general formula I as claimed in claims 1 to 9 and their pharmaceutically usable salts, tautomers and stereoisomers, including mixtures thereof in all ratios, which process is characterized in that,

a) a compound of the general formula II,

wherein X, Y and R¹Having the meaning given in claim 1,

reacting with a compound shown in a general formula III,

R-CO-L III

in which R has the meaning given in claim 1, L represents Cl, Br, I or a free OH group or a functionally modified OH group which is reactive,

alternatively, the first and second electrodes may be,

and/or converting a base or acid of formula I into a salt thereof.

11. A medicament comprising at least one compound of the general formula I according to claims 1 to 9 and/or pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, and optionally excipients and/or adjuvants.

12. Compounds according to claims 1-9 and pharmaceutically usable salts, tautomers and stereoisomers thereof, including mixtures thereof in various ratios, for use in the treatment of the following diseases: cancer, septic shock, Primary Open Angle Glaucoma (POAG), hyperplasia, rheumatoid arthritis, psoriasis, atherosclerosis, retinopathy, osteoarthritis, endometriosis, chronic inflammation and/or neurodegenerative diseases.

13. Compounds of the general formula I according to claims 1 to 9 and/or their physiologically acceptable salts, tautomers and stereoisomers for the use in the treatment of tumors, wherein a therapeutically effective amount of a compound of the general formula I is combined with substances within the group: group 1) of: estrogen receptor modulators, group 2): androgen receptor modulators, group 3): retinoid receptor modulators, group 4): cytotoxic agent, group 5): antiproliferative agents, group 6): prenyl protein transferase inhibitors, group 7): HMG-CoA reductase inhibitors, group 8): HIV protease inhibitors, group 9): reverse transcriptase inhibitors, and group 10): other angiogenesis inhibitors.

14. Compounds of the general formula I according to claims 1 to 9 and/or their physiologically acceptable salts, tautomers and stereoisomers for the use in the treatment of tumors, wherein a therapeutically effective amount of a compound of the general formula I is used in combination with radiotherapy and substances within the group: group 1) of: estrogen receptor modulators, group 2): androgen receptor modulators, group 3): retinoid receptor modulators, group 4): cytotoxic agent, group 5): antiproliferative agents, group 6): prenyl protein transferase inhibitors, group 7): HMG-CoA reductase inhibitors, group 8): HIV protease inhibitors, group 9): reverse transcriptase inhibitors, and group 10): other angiogenesis inhibitors.