HK1151291B - 2-benzylpyridazinone derivatives as met kinase inhibitors - Google Patents
2-benzylpyridazinone derivatives as met kinase inhibitors Download PDFInfo
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Description
Background
The object of the present invention was to find new compounds with valuable properties, in particular those which can be used for the preparation of medicaments.
The present invention relates to compounds and the use of compounds in which the inhibition, regulation and/or modulation of signal transduction by kinases, in particular tyrosine kinases and/or serine/threonine kinases, plays a role, to pharmaceutical compositions comprising these compounds and to the use of these compounds for the treatment of kinase-induced diseases.
In particular, the present invention relates to compounds and uses of compounds in which inhibition, regulation and/or modulation of signal transduction by Met kinase plays a role.
One of the major mechanisms for achieving cellular regulation is through transmembrane extracellular signal transduction, which in turn further regulates the biochemical pathways within the cell. Protein phosphorylation represents a process by which intracellular signals propagate from molecule to molecule, ultimately leading to a cellular response. These signal transduction cascades are highly regulated and often overlap, as can be seen by the presence of multiple protein kinases as well as phosphatases. Protein phosphorylation occurs primarily at serine, threonine or tyrosine residues, and protein kinases have therefore been classified by the specificity of their phosphorylation sites, i.e., serine/threonine kinases and tyrosine kinases. Since phosphorylation is a ubiquitous process within cells and cellular phenotypes are largely influenced by the activity of these pathways, it is currently believed that many disease states and/or diseases can be attributed to aberrant activation or functional mutations of molecular components of the kinase cascade. Therefore, much attention has been devoted to the characterization of these proteins and compounds that modulate their activity (for a review see: Weinstein-Oppenheimer et al, Pharma. &. Therap., 2000, 88, 229-279).
Berthou et al describe the role of receptor tyrosine kinase Met in human neoplasia and the possibility of inhibiting HGF (hepatocyte growth factor) -dependent Met activation in Oncogene, Vol.23, No. 31, p.5387-5393 (2004). The inhibitor described therein SU11274 (a pyrrole-indoline compound) may be useful against cancer. J.G Christensen et al describe another Met kinase inhibitor for cancer therapy in cancer Res.2003, 63(21), 7345-55. Hov et al, Clinical Cancer Research, Vol.10, 6686-6694(2004), reported another tyrosine kinase inhibitor for use against Cancer. The compound PHA-665752 (an indole derivative) acts against the HGF receptor c-Met. In addition, HGF and Met have been reported to have a large effect on the malignant process of various forms of cancer, such as multiple myeloma.
Therefore, there is a need to synthesize small compounds that specifically inhibit, regulate and/or modulate signal transduction by tyrosine kinases and/or serine/threonine kinases (particularly Met kinase), which is an object of the present invention.
The compounds of the invention and their salts have been found to possess very valuable pharmacological properties while being well tolerated.
The invention particularly relates to compounds of formula (I) capable of inhibiting, modulating and/or regulating Met kinase signal transduction, compositions comprising these compounds and methods of their use in mammals for the treatment of Met kinase-induced diseases and disorders (complains) such as angiogenesis, cancer, tumour formation, growth and spread, arteriosclerosis, ocular diseases such as age-induced macular degeneration, choroidal neovascularization and diabetic retinopathy, inflammatory diseases, arthritis, thrombosis, fibrosis, glomerulonephritis, neurodegeneration, psoriasis, restenosis, wound healing, graft rejection, metabolic diseases and diseases of the immune system, as well as autoimmune diseases, cirrhosis, diabetes and vascular diseases, as well as instability (instability) and permeability (trimeability), and the like
Solid tumors, particularly rapidly growing tumors, including monocytic leukemia, brain cancer, genitourinary tract cancer, lymphatic system cancer, stomach cancer, laryngeal cancer, and lung disorders, including lung adenocarcinoma and small cell lung cancer, can be treated with Met kinase inhibitors.
The present invention relates to methods of modulating, modulating or inhibiting Met kinase for the prevention and/or treatment of diseases associated with deregulated or disturbed Met kinase activity. In particular, the compounds of formula (I) can also be used to treat certain forms of cancer. The compounds of formula (I) may also be used to provide additive or synergistic effects in certain existing cancer chemotherapies, and/or can be used to restore the efficacy of certain existing cancer chemotherapies and radiation therapies.
The compounds of formula (I) can also be used to isolate and study Met kinase activity or expression. Furthermore, they are particularly suitable for use in diagnostic methods for diseases associated with disregulation or disturbance of Met kinase activity.
The compounds of the invention have been shown to have an antiproliferative effect in vivo in a xenograft tumor model. The compounds of the invention are administered to a patient with a hyperproliferative disease, for example, to inhibit tumor growth, reduce inflammation associated with lymphoproliferative diseases, inhibit graft rejection or nerve damage due to tissue repair, and the like. The compounds of the invention are suitable for prophylactic or therapeutic purposes. The term "treatment" as used herein refers to both prevention of disease and treatment of an already existing condition. Prevention of proliferation, e.g., prevention of tumor growth, prevention of metastatic growth, reduction of restenosis associated with cardiovascular surgery, etc., is achieved by administering a compound of the present invention prior to the development of overt disease. Alternatively, the compounds are used to treat an ongoing disease by stabilizing or ameliorating the clinical symptoms of the patient.
The host or patient may include any mammalian species, for example primates, particularly humans; rodents, including mice, rats, and hamsters; a rabbit; horses, cattle, dogs, cats, etc. Experimental studies have focused on animal models that provide a model for the treatment of human diseases.
The sensitivity of a particular cell to treatment with a compound of the invention can be determined by in vitro assays. Typically, the cell culture is mixed with varying concentrations of a compound of the invention for a period of time sufficient to allow the active ingredient to induce cell death or inhibit migration, typically about 1 hour to 1 week. In vitro testing may be performed with cultured cells from a biopsy sample. The remaining viable cells after treatment were then counted.
The dosage will vary depending upon the particular compound employed, the particular disease, the patient's condition, and the like. The therapeutic dose is generally sufficient to significantly reduce the undesirable cell population in the target tissue while maintaining the viability of the patient. Treatment generally continues until a significant reduction occurs, e.g., a reduction in cell burden (cell burden) of at least about 50%, and may continue until substantially no more undesired cells are detected in the body.
In order to identify signal transduction pathways and to detect interactions between different signal transduction pathways, different scientists have developed suitable 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 determine certain stages in the signal transduction cascade, interacting compounds may be utilized in order to modulate the signal (e.g., Stephens et al, Biochemical j., 2000, 351, 95-105). The compounds of the invention may also be used as reagents for testing kinase-dependent signal transduction pathways in animal and/or cell culture models or in clinical conditions as described herein.
The determination of kinase activity is a technique well known to those skilled in the art. General test systems for determining kinase activity using substrates such as histones (e.g.Alessi et al, FEBS Lett.1996, 399, 3, page 333-338) or basic myelin proteins are described in the literature (e.g.Campos-Gonz lez, R. and Glenney, Jr., J.R.1992, J.biol.chem.267, page 14535).
For the identification of kinase inhibitors, a variety of assay systems are available. In the scintillation proximity assay (Sorg et al, j.of. Biomolecular Screening, 2002, 7, 11-19) and flash plate assay, gamma ATP is used to determine the radioactive phosphorylation of proteins or peptides as substrates. In the presence of the inhibitory compound, a reduced radioactive signal or no radioactive signal at all can be detected. Furthermore, uniform time-resolved fluorescence resonance energy transfer (HTR-FRET) and Fluorescence Polarization (FP) techniques are also suitable for use as assay methods (Sills et al, J.of Biomolecular Screening, 2002, 191-214).
Other non-radioactive ELISA assays use specific phospho-antibodies (phospho-ABs). phospho-AB binds only phosphorylated substrates. This binding can be detected by chemiluminescence using peroxidase-conjugated anti-sheep secondary antibodies (Ross et al, 2002, biochem.j.).
There are many diseases associated with dysregulation of cell proliferation and cell death (apoptosis). Disorders of interest include, but are not limited to, those below. The compounds of the invention are suitable for use in the treatment of a variety of conditions in which there is proliferation and/or migration of smooth muscle cells and/or inflammatory cells into the vascular lining resulting in restricted blood flow through the vessel, for example in the case of neointimal occlusive lesions. Occlusive graft vascular diseases of interest include atherosclerosis, post-transplant coronary vascular disease, venous graft stenosis, peri-anastomotic restenosis (peri-arterial restenosis), restenosis following angioplasty or stent placement, and the like.
Prior Art
Other pyridazine derivatives as MET kinase inhibitors are disclosed in WO 2007/065518.
Thiadiazinones (Thiadiazinones) are disclosed in DE19604388, WO2003/037349, WO2007/057093 or WO 2007/057092.
Dihydropyridazinones against cancer are disclosed in WO 03/037349a 1.
Other pyridazine compounds for the treatment of diseases of the immune system, ischemic diseases and inflammatory diseases are known from EP 1043317 a1 and EP 1061077 a 1.
EP 0738716 a2 and EP 0711759B 1 disclose other dihydropyridazinones and pyridazinones as fungicides and insecticides.
Pyridazinones as cardiotonic ingredients are described in US4,397,854.
JP 57-95964 discloses other pyridazinone compounds.
Summary of the invention
The present invention relates to compounds of formula (I) and pharmaceutically acceptable salts, tautomers and stereoisomers thereof (including mixtures thereof in all ratios):
wherein:
R1represents a group of formula (I) wherein R represents H or A,
R2represents an unsaturated, saturated or aromatic 5-or 6-membered heterocyclic ring having 1 to 4N, O and/or S atoms, which heterocyclic ring may be unsubstituted or mono-, di-or trisubstituted by: hal, A, [ C (R) ]3)2】nOR3、N＝CR3N(R3)2、SR3、NO2、CN、COOR3、CON(R3)2、NR3COA、NR3SO2A、SO2N(R3)2、S(O)mA、【C(R3)2]nN(R3)2、【C(R3)2]nHet、O【C(R3)2】nN(R3)2、O【C(R3)2】nHet、S【C(R3)2】nN(R3)2、S【C(R3)2】nHet、NR3【C(R3)2】nN(R3)2、NR3【C(R3)2]nHet、NHCON(R3)2、NHCONH【C(R3)2】nN(R3)2、NHCONH【C(R3)2】nHet、【C(R3)2】nNHCO【C(R3)2】nN(R3)2、【C(R3)2]nNHCO【C(R3)2]nHet、CON(R3)2、CONR3【C(R3)2】nN(R3)2、CONR3【C(R3)2]nNR3COOA、CONR3【C(R3)2]nOR3、CONR3【C(R3)2】nHet, COHet, COA and/or ═ O (carbonyl oxygen),
R3represents a group of formula (I) wherein R represents H or A,
R4、R4' each independently of the other represents H, Hal, A, OR3、CN、COOR3、CON(R3)2、NR3COA、NR3SO2A、SO2N(R3)2Or S (O)mA
Ar represents phenyl, naphthyl or biphenyl, each of which is unsubstituted or mono-, di-or trisubstituted by: hal, A, [ C (R) ]3)2】nOR3、【C(R3)2】nN(R3)2、SR3、NO2、CN、COOR3、CON(R3)2、NR3COA、NR3SO2A、SO2N(R3)2、S(O)mA、CO-Het、Het、O【C(R3)2】nN(R3)2、O【C(R3)2】nHet、NHCOOA、NHCON(R3)2、NHCOO【C(R3)2】nN(R3)2、NHCOO【C(R3)2】nHet、NHCONH【C(R3)2】nN(R3)2、NHCONH【C(R3)2】nHet、OCONH【C(R3)2】nN(R3)2、OCONH【C(R3)2】nHet、CONR3【C(R3)2】nN(R3)2、CONR3【C(R3)2】nHet and/or COA, and/or,
het represents a mono-, bi-or tricyclic saturated, unsaturated or aromatic heterocycle having 1 to 4N, O and/or S atoms, which heterocycle may be unsubstituted or mono-, di-or trisubstituted by: hal, A, [ C (R) ]3)2】nOR3、【C(R3)2】nN(R3)2、SR3、NO2、CN、COOR3、CON(R3)2、NR3COA、NR3SO2A、SO2N(R3)2、S(O)mA、COHet1、【C(R3)2】nHet1、O【C(R3)2】nN(R3)2、O【C(R3)2】nHet1、NHCOOA、NHCON(R3)2、NHCOO【C(R3)2]nN(R3)2、NHCOO【C(R3)2]nHet1、NHCONH【C(R3)2】nN(R3)2、NHCONH【C(R3)2]nHet1、OCONH【C(R3)2】nN(R3)2、OCONH【C(R3)2]nHet1、CO-Het1CHO, COA, ═ S, ═ NH, ═ NA and/or ═ O (carbonyl oxygen),
Het1represents a monocyclic saturated heterocycle having 1 to 2N and/or O atoms, which heterocycle may be mono-or disubstituted by A, OA, OH, Hal and/or ═ O (carbonyl oxygen),
a represents an unbranched or branched alkyl radical having 1 to 10 carbon atoms in which 1 to 7H atoms may be replaced by F and/or in which 1 or 2 non-adjacent CH groups2The radicals being selected from the group consisting of O, NH, S, SO2And/or CH ═ CH groups, or cycloalkyl groups having 3 to 7 carbon atoms,
hal represents F, Cl, Br or R,
m represents 0, 1 or 2,
n represents 0, 1, 2, 3 or 4.
The compounds of formula (I) are also meant to be pharmaceutically acceptable derivatives and solvates thereof.
The invention also relates to optically active forms (stereoisomers), enantiomers, racemates, diastereomers and hydrates and solvates of these compounds. The term solvate of a compound means that inert solvent molecules are adducted to the compound, which are formed due to their mutual attraction. Solvates are, for example, mono-or di-hydrates or alcoholates.
The term pharmaceutically acceptable derivatives means, for example, salts of the compounds of the invention, and also so-called prodrug compounds.
The term prodrug derivative means a compound of formula (I) which is modified with, for example, an alkyl or acyl group, a sugar or an oligopeptide and which is capable of being rapidly cleaved in vivo to form the effective compounds of the invention.
These also include biodegradable polymer derivatives of the compounds of the invention, such as, for example, int.j.pharm.11561-67 (1995).
The expression "effective amount" means the amount of a drug or pharmaceutically active ingredient that produces a biological or medical response in a tissue, system, animal or human that is, for example, sought or desired by a researcher or physician. Furthermore, the expression "therapeutically effective amount" denotes an amount which produces the following effect compared to the corresponding individual not receiving this amount: improved treatment, healing, prevention or elimination of a disease, syndrome, condition, disorder, or side effect, or alternatively, reduced progression of a disease, disorder, or condition. The expression "therapeutically effective amount" also includes amounts that are effective in terms of enhancing normal physiological function.
The invention also relates to the use of mixtures of compounds of the formula (I), for example mixtures of two diastereomers in a ratio of 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, 1: 10, 1: 100 or 1: 1000. Mixtures of the stereoisomeric compounds are particularly preferred.
The present invention relates to compounds of formula (I) and salts thereof, to a process for the preparation of compounds of formula (I) according to claims 1 to 10 and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, characterized in that:
a) reacting a compound of formula (II):
wherein R is1As defined in claim 1, the first and second,
with a compound of the formula (III),
wherein R is2、R3、R4And R4' as defined in claim 1, and
l represents Cl, Br, I or a free or reactive functional group-modified OH group,
or
b) The group R is prepared by the following reaction2Conversion to another group R2：
i) The heterocyclic ring is arylated to form the heterocyclic ring,
ii) acylation or alkylation of the amino group,
iii) etherification of the hydroxyl groups,
or
c) It is released from its functional group derivative by treatment with a solvolytic or hydrogenolytic agent,
and/or
Converting a base or acid of formula (I) into one of its salts.
Unless otherwise explicitly stated, the radicals R in the context1、R2、R3、R4、R4' is as defined for formula (I).
A represents an alkyl group, which is unbranched (linear) or branched, having 1, 2, 3, 4, 5, 6, 7,8, 9 or 10C atoms. A preferably represents methyl, furthermore ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, furthermore 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, furthermore preferably, for example, trifluoromethyl.
A very particularly preferably represents alkyl having 1, 2, 3, 4, 5 or 6C atoms, preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, trifluoromethyl, pentafluoroethyl or 1, 1, 1-trifluoroethyl.
Cyclic alkyl (cycloalkyl) preferably denotes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
Ar represents, for example, o-, m-or p-tolyl, o-, m-or p-ethylphenyl, o-, m-or p-propylphenyl, o-, m-or p-isopropylphenyl, o-, m-or p-tert-butylphenyl, o-, m-or p-hydroxyphenyl, o-, m-or p-nitrophenyl, o-, m-or p-aminophenyl, o-, m-or p- (N-methylamino) phenyl, o-, m-or p- (N-methylaminocarbonyl) phenyl, o-, m-or p-acetylaminophenyl, o-, m-or p-methoxyphenyl, o-, m-or p-ethoxyphenyl, o-, m-or p-ethoxycarbonylphenyl, o-, m-or p- (N, N-dimethylamino) phenyl, o-, m-or p- (N, N-dimethylaminocarbonyl) phenyl, o-, m-or p- (N-ethylamino) phenyl, o-, m-or p- (N, N-diethylamino) phenyl, o-, m-or p-fluorophenyl, o-, m-or p-bromophenyl, o-, m-or p-chlorophenyl, o-, m-or p- (methylsulfonylamino) phenyl, o-, m-or p- (methylsulfonyl) phenyl, o-, m-or p-methylsulfanylphenyl, o-, m-or p-cyanophenyl, o-, m-or p-carboxyphenyl, o-, m-or p-methoxycarbonylphenyl, o-, m-or p-formylphenyl, o-, m-or p-acetylphenyl, o-, m-or p-aminosulfonylphenyl, o-, m-or p- (morpholin-4-ylcarbonyl) phenyl, o-, m-or p- (3-oxomorpholin-4-yl) phenyl, o-, m-or p- (piperidinylcarbonyl) phenyl, o-, m-or p- [ 2- (morpholin-4-yl) ethoxy ] phenyl o-, m-or p- [3- (N, N-diethylamino) propoxy ] phenyl, o-, m-or p- [3- (3-diethylaminopropyl) ureido ] phenyl, o-, m-or p- (3-diethylaminopropoxycarbonylamino) phenyl, furthermore, preferably 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, 3-, 2, 4-, or 3, 5-dichlorophenyl, 2, 5-, 2, 6-, 3, 4-or 3, 5-dibromophenyl, 2, 4-or 2, 5-dinitrophenyl, 2, 5-or 3, 4-dimethoxyphenyl, 3-nitro-4-chlorophenyl, 3-amino-4-chloro-, 2-amino-3-chloro-, 2-amino-4-chloro-, 2-amino-5-chloro-or 2-amino-6-chlorophenyl, 2-nitro-4-N, N-dimethylamino-or 3-nitro-4-N, N-dimethylaminophenyl, 2, 3-diaminophenyl, 2, 3, 4-), 2, 3, 5-, 2, 3, 6-, 2, 4, 6-or 3, 4, 5-trichlorophenyl, 2, 4, 6-trimethoxyphenyl, 2-hydroxy-3, 5-dichlorophenyl, p-iodophenyl, 3, 6-dichloro-4-aminophenyl, 4-fluoro-3-chlorophenyl, 2-fluoro-4-bromophenyl, 2, 5-difluoro-4-bromophenyl, 3-bromo-6-methoxyphenyl, 3-chloro-4-acetylaminophenyl, 3-fluoro-4-methoxyphenyl, 3-amino-6-methylphenyl, 3-chloro-4-acetylaminophenyl or 2, 5-dimethyl-4-chlorophenyl.
Ar also preferably represents phenyl, naphthyl or biphenyl, each of which is unsubstituted or substituted by Hal, CN, O [ C (R)3)2】nN(R3)2、CONR3【C(R3)2】nN(R3)2And/or CONR3［C(R3)2]nHet is mono-, di-or tri-substituted.
Het represents, irrespective of the additional substitution, for example 2-or 3-furyl, 2-or 3-thienyl, 1-, 2-or 3-pyrrolyl, 1-, 2-, 4-or 5-imidazolyl, 1-, 3-, 4-or 5-pyrazolyl, 2-, 4-or 5-oxazolyl, 3-, 4-or 5-isoxazolyl, 2-, 4-or 5-thiazolyl, 3-, 4-or 5-isothiazolyl, 2-, 3-or 4-pyridyl, 2-, 4-, 5-or 6-pyrimidinyl, furthermore preferably 1, 2, 3-triazol-1-, -4-or-5-yl, optionally substituted phenyl, optionally, 1, 2, 4-triazol-1-, -3-or 5-yl, 1-or 5-tetrazolyl, 1, 2, 3-oxadiazol-4-or-5-yl, 1, 2, 4-oxadiazol-3-or-5-yl, 1, 3, 4-thiadiazol-2-or-5-yl, 1, 2, 4-thiadiazol-3-or-5-yl, 1, 2, 3-thiadiazol-4-or-5-yl, 3-or 4-pyridazinyl, pyrazinyl, 1-, 2-, 3-, 4-, 5-, 6-or 7-indolyl, 4-or 5-isoindolyl, indazolyl, tetrazolyl, thiadiazol-2-or-5-yl, thiadiazol-3-, or-5-yl, 1, 2, 4-, 5-, 6-or 7-indolyl, 1-, 2-, 4-or 5-benzimidazolyl, 1-, 3-, 4-, 5-, 6-or 7-benzopyrazolyl, 2-, 4-, 5-, 6-or 7-benzoxazolyl, 3-, 4-, 5-, 6-or 7-benzisoxazolyl, 2-, 4-, 5-, 6-or 7-benzothiazolyl, 2-, 4-, 5-, 6-or 7-benzisothiazolyl, 4-, 5-, 6-or 7-benzo-2, 1, 3-oxadiazolyl, 2-, 3-, 4-, 5-, 6-, 7-or 8-quinolyl, and pharmaceutically acceptable salts thereof, 1-, 3-, 4-, 5-, 6-, 7-or 8-isoquinolinyl, 3-, 4-, 5-, 6-, 7-or 8-cinnolinyl, 2-, 4-, 5-, 6-, 7-or 8-quinazolinyl, 5-or 6-quinoxalinyl, 2-, 3-, 5-, 6-, 7-or 8-2H-benzo-1, 4-oxazinyl, preference is also given to 1, 3-benzodioxol-5-yl, 1, 4-benzodioxan-6-yl, 2, 1, 3-benzothiadiazol-4-or-5-yl, 2, 1, 3-benzoxadiazol-5-yl or dibenzofuranyl.
The heterocyclic group may also be partially or fully hydrogenated.
Het may therefore also represent, for example, 2, 3-dihydro-2-, -3-, -4-or-5-furyl, 2, 5-dihydro-2-, -3-, -4-or 5-furyl, tetrahydro-2-or-3-furyl, 1, 3-dioxolan-4-yl, tetrahydro-2-or-3-thienyl, 2, 3-dihydro-1-, -2-, -3-, -4-or-5-pyrrolyl, 2, 5-dihydro-1-, -2-, -3-, 4-or-5-pyrrolyl, irrespective of the further substitution, 1-, 2-or 3-pyrrolidinyl, tetrahydro-1-, 2-or 4-imidazolyl, 2, 3-dihydro-1-, -2-, -3-, -4-or 5-pyrazolyl, tetrahydro-1-, -3-or 4-pyrazolyl, 1, 4-dihydro-1-, -2-, -3-or 4-pyridyl, 1, 2, 3, 4-tetrahydro-1-, -2-, -3-, -4-, -5-or 6-pyridyl, 1-, 2-, 3-or 4-piperidyl, 2-, 3-or 4-oxazolinyl, tetrahydro-2-, -3-or-4-pyranyl, 1, 4-dioxanyl, 1, 3-dioxan-2-, -4-or-5-yl, hexahydro-1-, -3-or-4-pyridazinyl, hexahydro-1-, -2-, -4-or-5-pyrimidinyl, 1-, 2-or 3-piperazinyl, 1, 2, 3, 4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7-or-8-quinolinyl, 1, 2, 3, 4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7-or-8-isoquinolinyl, 2-, 3-, 5-, 6-, 7-or 8-3, 4-dihydro-2H-benzo-1, 4-oxazinyl, further preferably 2, 3-methylenedioxyphenyl, 3, 4-methylenedioxyphenyl, 2, 3-ethylenedioxyphenyl, 3, 4- (difluoromethylenedioxy) phenyl, 2, 3-dihydrobenzofuran-5-or 6-yl, 2, 3- (2-oxomethylenedioxy) phenyl, or further 3, 4-dihydro-2H-1, 5-benzodioxepin-6-or-7-yl, furthermore preferably 2, 3-dihydrobenzofuranyl, 2, 3-dihydro-2-oxofuranyl, 3, 4-dihydro-2-oxo-1H-quinazolinyl, 2, 3-dihydrobenzoxazolyl, 2-oxo-2, 3-dihydrobenzoxazolyl, 2-3-dihydrobenzimidazolyl, 1, 3-dihydroindole, 2-oxo-1, 3-dihydroindole or 2-oxo-2, 3-dihydrobenzimidazolyl.
Het preferably represents a monocyclic ring having 1 to 4N, O and/or S atomsSaturated, unsaturated or aromatic heterocycles, which may be unsubstituted or substituted by A and/or [ C (R)3)2】nHet1Mono-, di-or tri-substituted.
Het particularly preferably represents piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridazinyl or pyrazinyl, each of which is substituted by A and/or [ C (R)3)2】nHet1Single substitution.
Het1Preferably represents a monocyclic saturated heterocycle having 1 to 2N and/or O atoms, which may be mono-or disubstituted by a and/or ═ O (carbonyl oxygen).
Het1Preferably represents pyrrolidine, piperidine, piperazine or morpholine, each of which is unsubstituted or mono-or disubstituted by a and/or ═ O (carbonyl oxygen).
R1Preferably represents cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methyl, ethyl, propyl or isopropyl and may also be H.
At R2Unsaturated, saturated or aromatic 6-membered heterocycles having 1 to 4N and/or O atoms in the definitions have the following definitions, for example: furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl or thiadiazolyl.
R2Preferably represents an unsaturated or aromatic 5-or 6-membered heterocycle having 1 to 4N and/or O atoms, which may be unsubstituted or substituted by Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2】nHet、O【C(R3)2]nN(R3)2And/or O [ C (R)3)2】nHet is mono-, di-or tri-substituted.
R2Particularly preferably represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl or thiadiazolyl, each of which is Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2】nHet、O【C(R3)2】nN(R3)2Or O [ C (R)3)2】nHet
R3Preferably represents H, methyl, ethyl or propyl, very particularly preferably H.
R4、R4' preferably represents H.
Hal preferably represents F, Cl or Br, but may also be I, particularly preferably F or Cl.
In the present invention, all groups appearing more than once may be the same or different, i.e. independently of each other.
The compounds of formula (I) may have one or more chiral centres and may therefore occur in a number of stereoisomeric forms. Formula (I) encompasses all of these forms.
The invention therefore relates in particular to compounds of the formula (I) in which at least one of the radicals mentioned has one of the preferred meanings indicated above. Certain preferred groups of compounds can be represented by the following subformulae (I) a to Ik, which conform to formula (I) and in which the groups not specified in more detail have the meanings given for formula (I), but in which:
in Ia, R2Represents an unsaturated or aromatic 5-or 6-membered heterocycle having 1 to 4N and/or O atoms which may be unsubstituted or substituted by Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2］nHet、O [C(R3)2】nN(R3)2And/or O [ C (R)3)2】nHet is mono-, di-or tri-substituted;
in Ib, R4、R4' represents H;
in Ic Het represents a monocyclic saturated, unsaturated or aromatic heterocycle having 1 to 4N, O and/or S atoms which may be unsubstituted or substituted by A and/or [ C (R)3)2］nHet1Mono-, di-or tri-substituted;
in Id, A represents an unbranched or branched alkyl radical having 1 to 8 carbon atoms, in which 1 to 7H atoms may be replaced by F and/or Cl, or A represents a cycloalkyl radical having 3 to 7 carbon atoms;
in Ie, R3Represents H, methyl, ethyl or propyl;
in If, Het1Represents a monocyclic saturated heterocycle having 1 to 2N and/or O atoms, which may be mono-or disubstituted by a and/or ═ O (carbonyl oxygen);
in Ig, R2Represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl or thiadiazolyl, each of which is Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2］nHet、O【C(R3)2】nN(R3)2Or O [ C (R)3)2】nHet is monosubstituted;
in Ih Het represents piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridazinyl or pyrazinyl, each of which is substituted by a or [ C (R)3)2】nHet1Monosubstitution;
in Ii, Het1Represents pyrrolidine, piperidine, piperazine or morpholine, each of which is unsubstituted or mono-or disubstituted by a and/or ═ O (carbonyl oxygen);
in Ij:
R1represents a group of formula (I) wherein R represents H or A,
R2represents an unsaturated or aromatic 5-or 6-membered heterocycle having 1 to 4N and/or O atoms which may be unsubstituted or substituted by Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2】nHet、O【C(R3)2】nN(R3)2And/or O [ C (R) ]3)2】nHet is mono-, di-or trisubstituted,
R3represents H, methyl, ethyl or propyl,
R4、R4' represents a hydrogen atom in the molecule of H,
het represents a monocyclic saturated, unsaturated or aromatic heterocycle having 1 to 4N, O and/or S atoms which may be unsubstituted or substituted by A and/or [ C (R)3)2】nHet1Mono-, di-or tri-substituted,
Het1represents a monocyclic saturated heterocycle having 1 to 2N and/or O atoms, which may be mono-or disubstituted by A and/or ═ O (carbonyl oxygen),
a represents unbranched or branched alkyl having 1 to 8 carbon atoms, in which 1 to 7H atoms may be replaced by F and/or Cl, or represents cycloalkyl having 3 to 7 carbon atoms,
hal represents F, Cl, Br or I,
n represents 0, 1, 2, 3 or 4;
in Ik:
R1represents a group of formula (I) wherein R represents H or A,
R2represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl or thiadiazolyl, each of which is Hal, A, [ C (R)3)2】nN(R3)2、【C(R3)2］nHet、O【C(R3)2】nN(R3)2Or O [ C (R)3)2】nHet is singly substituted by a substituent group (Het),
R3represents H, methyl, ethyl or propyl,
R4、R4' represents a hydrogen atom in the molecule of H,
het represents piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridazinyl or pyrazinyl, each of which is substituted by A or [ C (R)3)2】nHet1The process is a single substitution process,
Het1represents pyrrolidine, piperidine, piperazine or morpholine, each of which is unsubstituted or mono-or disubstituted by A and/or ═ O (carbonyl oxygen),
a represents unbranched or branched alkyl having 1 to 8 carbon atoms, in which 1 to 7H atoms may be replaced by F and/or Cl, or represents cycloalkyl having 3 to 7 carbon atoms,
hal represents F, Cl, Br or I,
n represents 0, 1, 2, 3 or 4;
and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios.
Furthermore, the compounds of formula (I) and the starting materials for their preparation can be prepared according to well-known methods, for example as described in the literature (for example in standard textbooks such as Houben-Weyl, Methoden der organischen Chemie [ methods of organic chemistry ], Georg-Thieme-Verlag, Stuttgart), precisely under reaction conditions which are known and suitable for the reaction in question. Well-known variations not mentioned in more detail herein may also be employed.
The starting compounds of the formulae (II) and (III) are generally known. However, if they are new, they can be prepared by a well-known method.
The pyridazinones of formula (II) used may generally be prepared according to the procedures of W.J. coats, A.McKillop, Synthesis, 1993, 334-342 if not commercially available.
The compounds of formula (I) can preferably be prepared by reacting a compound of formula (II) with a compound of formula (III).
In the compounds of the formula (III), L preferably represents Cl, Br, I or a free or reactively modified OH group, for example an active ester, an imidazole anion (imidazolide) or an alkylsulphonyloxy group having 1 to 6C atoms, preferably a methanesulphonyloxy or trifluoromethanesulphonyloxy group, or an arylsulphonyloxy group having 6 to 10C atoms, preferably a phenyl-or p-toluenesulphonyloxy group.
The reaction is generally carried out in the presence of an acid-binding agent, preferably an organic base such as DIPEA, triethylamine, dimethylaniline, pyridine or quinoline.
It may also be advantageous to add a hydroxide, carbonate or bicarbonate of an alkali or alkaline earth metal, preferably potassium, sodium, calcium or cesium, or another weak acid salt of an alkali or alkaline earth metal.
Depending on the conditions employed, the reaction time is from a few minutes to 14 days and the reaction temperature is from about-30 ℃ to 140 ℃, usually from-10 ℃ to 90 ℃, in particular from about 0 ℃ to about 70 ℃.
Examples of suitable inert solvents are: hydrocarbons, such as hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons, such as trichloroethylene, 1, 2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; alcohols, such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; ethers such as diethyl ether, diisopropyl ether, Tetrahydrofuran (THF) or dioxane; glycol ethers, such as ethylene glycol monomethyl or monoethyl ether, 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 said solvents. Particular preference is given to acetonitrile, dichloromethane and/or DMF.
The reaction of the compound of the formula (II) with the compound of the formula (III) in which L represents OH is preferably carried out under Mitsunobu reaction conditions by adding, for example, triphenylphosphine and a dialkyl azodicarboxylate. THF is preferably used as the solvent.
Compounds of formula (I) may also be prepared by reacting the radical R2Conversion to another group R2And are obtained, for example, by arylating heterocycles in a Suzuki reaction.
The compounds of formula (I) may also be obtained by solvolysis, in particular hydrolysis, or by liberation from their functional derivatives by hydrogenolysis.
Preferred starting materials for solvolysis or hydrogenolysis are those containing the corresponding protected amino and/or hydroxyl group instead of one or more free amino and/or hydroxyl groups, preferably those carrying an amino-protecting group instead of an H atom bound to an N atom, e.g. those corresponding to formula (I) but containing an NHR 'group (wherein R' is an amino-protecting group, e.g. BOC or CBZ) instead of NH2Starting from a radical.
Also preferred are starting materials which carry H atoms of a hydroxy-protecting group other than hydroxy, such as those corresponding to formula (I) but which contain an R 'O-phenyl group (where R' is a hydroxy-protecting group) other than a hydroxyphenyl group.
In the starting molecule, there may also be a plurality of (identical or different) protected amino and/or hydroxyl groups. If the protecting groups present are different from one another, they can be cleaved off selectively in many cases.
The term "amino-protecting group" is known as a generic term and refers to a group that is suitable for protecting (blocking) an amino group from chemical reactions, but is readily removable after the desired chemical reaction at other positions in the molecule has taken place. Typical of such groups are in particular unsubstituted or substituted acyl, aryl, aralkyloxymethyl or aralkyl groups. Because the amino-protecting groups are removed after the desired reaction (or reaction step), their type and size are not particularly important; however, those having 1 to 20 (particularly 1 to 8) carbon atoms are preferred. In the present process, the term "acyl" is to be understood in its broadest sense. It includes acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids, especially alkoxycarbonyl, aryloxycarbonyl, especially 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 toluoyl (tolyl); aryloxyalkanoyl, such as POA; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, 2, 2, 2-trichloroethoxycarbonyl, BOC and 2-iodoethoxycarbonyl; aralkoxycarbonyl, such as CBZ ("carbobenzoxy"), 4-methoxybenzyloxycarbonyl, and FMOC; arylsulfonyl groups such as Mtr, Pbf, and Pmc. Preferred amino-protecting groups are BOC and Mtr, and also include CBZ, Fmoc, benzyl and acetyl.
The term "hydroxy-protecting group" is also a known generic term referring to a group that is suitable for protecting a hydroxy group from chemical reactions, but which is readily removable after the desired chemical reaction at other locations in the molecule has taken place. Typical such groups are the above unsubstituted or substituted aryl, aralkyl or acyl groups, also including alkyl groups. The nature and size of the hydroxy-protecting groups are not particularly important as they are removed after the desired chemical reaction or reaction step has been carried out; preference is given to radicals having from 1 to 20, in particular (1 to 10), carbon atoms. Examples of hydroxy-protecting groups are, inter alia, tert-butoxycarbonyl, benzyl, p-nitrobenzoyl, p-toluenesulfonyl, tert-butyl and acetyl, of which benzyl and tert-butyl are particularly preferred. The COOH groups in aspartic and glutamic acids are preferably protected in the form of their tert-butyl esters (e.g., Asp (OBut)).
The compounds of formula (I) may be liberated from their functional derivatives (depending on the protecting group used), for example by using strong acids, preferably TFA or perchloric acid, but also other strong inorganic acids, for example hydrochloric acid or sulfuric acid, strong organic carboxylic acids, for example trichloroacetic acid, or sulfonic acids, for example benzenesulfonic acid or p-toluenesulfonic acid. Other inert solvents may be present, but are not always necessary. Suitable inert solvents are preferably organic solvents, such as carboxylic acids, for example acetic acid; ethers such as tetrahydrofuran or dioxane; amides, such as DMF; halogenated hydrocarbons such as dichloromethane; and also alcohols, such as methanol, ethanol or isopropanol; and water. Mixtures of the above solvents are also suitable. Preferably TFA is used in excess without the addition of further solvents, the perchloric acid preferably being a mixture of acetic acid and 70% perchloric acid in 9: 1 in the form of a mixture. The reaction temperature for the cleavage is preferably from about 0 to about 50 deg.C, preferably from 15 to 30 deg.C (room temperature).
The BOC, Obut, Pbf, Pmc and Mtr groups can be cleaved off, for example, preferably with TFA in dichloromethane or with about 3-5N HCl in dioxane at 15-30 deg.C, and the FMOC groups can be cleaved off with about 5-50% dimethylamine, diethylamine or piperidine in DMF at 15-30 deg.C.
Trityl groups may be used to protect the amino acids histidine, asparagine, glutamine and cysteine. Depending on the desired end product, these can be cleaved off with TFA/10% thiophenol, and the trityl group can be cleaved off from all the amino acids mentioned above; when TFA/anisole or TFA/thioanisole is used, only the trityl group of His, Asn and Gln is cleaved off, while it remains on the Cys side chain.
Pbf (pentamethylbenzofuranyl) may be used to protect Arg. It can be removed using, for example, TFA in dichloromethane.
The hydrogenolytically removable protecting group (e.g. CBZ or benzyl) can be cleaved off, for example, by treatment with hydrogen in the presence of a catalyst (e.g. a noble metal catalyst, such as palladium, which is preferably on a support, e.g. on carbon). Suitable solvents herein are indicated as solvents, for example, alcohols (such as methanol or ethanol) or amides (such as DMF). Hydrogenolysis is generally carried out at a temperature of about 0 to 100 ℃ and a pressure of about 1 to 200bar, preferably at 20 to 30 ℃ and 1 to 1 obar. Hydrogenolysis of the CBZ group can be carried out, for example, on 5-10% Pd/C in methanol or with ammonium formate (instead of hydrogen) on Pd/C in methanol/DMF at 20-30 ℃.
Pharmaceutically acceptable salts and other forms
The compounds of the present invention can be used in their final non-salt form. In another aspect, the invention also encompasses the use of these compounds in the form of pharmaceutically acceptable salts, which can be derived from a variety of organic and inorganic acids and bases according to methods known in the art. The pharmaceutically acceptable salt forms of the compounds of formula (I) are mostly prepared by conventional methods. If the compound of formula (I) contains a carboxyl group, one of its suitable salts may be formed by reacting the compound with a suitable base to give the corresponding base addition salt. Such bases are, 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 potassium ethoxide and sodium propoxide; various organic bases, such as piperidine, diethanolamine and N-methyl-glutamine. Also included are aluminum salts of the compounds of formula (I). For certain compounds of formula (I), acid addition salts may be formed by treating such compounds with pharmaceutically acceptable organic and inorganic acids, for example hydrogen halides, such as hydrogen chloride, hydrogen bromide or hydrogen iodide; other inorganic acids and their corresponding salts, such as sulfates, nitrates or phosphates, etc.; alkyl-and monoaryl-sulfonates, such as ethanesulfonate, toluenesulfonate and benzenesulfonate; 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, arginate, aspartate, benzoate, benzenesulfonate (phenylsulfonate), bisulfate, bisulfite, bromide, butyrate, camphorate, camphorsulfonate, caprylate, chloride, chlorobenzoate, citrate, cyclopentanepropionate, digluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, fumarate, galactarate (from mucic acid), galacturonate, glucoheptonate, gluconate, glutamate, glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate, lactobionate, malate, dihydrogensulfate, fumarate, and the like, Maleate, malonate, mandelate, metaphosphate, methanesulfonate, methylbenzoate, monohydrogenphosphate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, oleate, pamoate (palmoate), pectinate, persulfate, phenylacetate, 3-phenylpropionate, phosphate, phosphonate, phthalate, but this is not intended to be limiting.
Further, the base salt of the compound of the present invention includes aluminum salt, ammonium salt, calcium salt, copper salt, iron (iii) salt, iron (ii) salt, lithium salt, magnesium salt, manganese (iii) salt, manganese (ii) salt, potassium salt, sodium salt and zinc salt, but this is not meant to be limiting. Among the above salts, ammonium salts are preferred; alkali metal salt sodium and potassium salts, and alkaline earth metal salt calcium and magnesium salts. Salts of the compounds of formula (I) derived from pharmaceutically acceptable organic non-toxic bases include salts of: primary, secondary and tertiary amines, substituted amines, and also 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-glucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethanolamine, triethylamine, trimethylamine, tripropylamine, and tris (hydroxymethyl) methylamine (tromethamine), but this is not meant to be limiting.
Compounds of the present invention containing basic nitrogen-containing groups may be quaternized with materials such as: (C)1-C4) Alkyl halides such as methyl, ethyl, isopropyl and tert-butyl chloride, bromide and iodide; sulfuric acid di (C)1-C4) Alkyl esters such as dimethyl, diethyl and diamyl sulfate; (C)10-C18) Alkyl halides such as decyl, dodecyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; and aryl (C)1-C4) Alkyl halides, such as benzyl chloride and phenethyl bromide. Such salts are employed to prepare water-soluble and oil-soluble compounds of the invention.
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, sulfosalicylate, tartrate, thiomalate, tosylate and tromethamine.
Particularly preferred are the hydrochloride, dihydrochloride, hydrobromide, maleate, methanesulfonate, phosphate, sulfate and succinate salts.
Acid addition salts of basic compounds of formula (I) may be prepared by contacting the free base form with a sufficient amount of the desired acid to form a salt in a conventional manner. The free base can be regenerated by contacting the salt form with a base and isolating the free base in a conventional manner. The free base form differs from its corresponding salt form in some respects with respect to certain physical properties, such as solubility in polar solvents; however, for the purposes of the present invention, salts are otherwise comparable to their respective free base forms.
As mentioned above, the pharmaceutically acceptable base addition salts of the compounds of formula (I) are formed 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-glucamine and procaine.
Base addition salts of the acidic compounds of the present invention may be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form the salt. The free acid may be regenerated by contacting the salt form with an acid and separating the free acid in a conventional manner. The free acid form differs from its corresponding salt form in some way with respect to certain physical properties, such as solubility in polar solvents; however, for the purposes of the present invention, salts are otherwise comparable to their respective free acid forms.
If the compounds of the present invention contain more than one group capable of forming such pharmaceutically acceptable salts, multiple salts are also encompassed by the present invention. Typical multiple salt forms include, for example, bitartrate, diacetate, difumarate, meglumine, diphosphate, disodium salt, and trihydrochloride, although this is not intended to be limiting.
In the light of the above, it can be seen that the expression "pharmaceutically acceptable salt" in the present context is intended to mean an active ingredient comprising a compound of formula (I) in one of its salt forms, in particular if this salt form confers improved pharmacokinetic properties on the active ingredient compared to the free form of the active ingredient or any other salt form of the active ingredient used earlier. The pharmaceutically acceptable salt form of the active ingredient can also for the first time provide the active ingredient with desired pharmacokinetic properties not previously possessed by it, and can even have a positive effect on the pharmacodynamics of the active ingredient in terms of its in vivo therapeutic effect.
The invention also relates to medicaments comprising at least one compound of formula (I) and/or pharmaceutically acceptable derivatives, solvates and stereoisomers thereof, including mixtures thereof in all ratios, and optionally excipients and/or adjuvants.
Pharmaceutical formulations can be administered in unit dosage forms containing a predetermined amount of the active ingredient per unit dose. Such units may contain, for example, from 0.5mg to 1g (preferably from 1mg to 700mg, particularly preferably from 5mg to 100mg) of a compound of the invention, depending on the condition to be treated, the method of administration and the age, weight and condition of the patient, or the pharmaceutical preparations may be administered in unit dosage form containing a predetermined amount of active ingredient per unit dosage. Preferred unit dose formulations are those containing the daily dose or partial dose, or corresponding fraction thereof, of the active ingredient. In addition, such pharmaceutical preparations may be prepared by methods widely known in the pharmaceutical field.
The pharmaceutical formulations can be adapted for administration by any desired suitable method, 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 formulations may be prepared by all methods known in the art of pharmacy, for example by mixing the active ingredient with excipients or adjuvants.
Pharmaceutical formulations adapted for oral administration may be administered in the form of discrete units such as capsules or tablets; a powder or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or foam foods; or an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
Thus, for example, for oral administration in the form of a tablet or capsule, the active ingredient component can be combined with non-toxic pharmaceutically acceptable oral inert excipients such as ethanol, glycerol, water and the like. Powders may be prepared by comminuting the compound to a suitably fine size and mixing it with a pharmaceutical excipient (e.g. an edible carbohydrate such as starch or mannitol) which is comminuted in a similar manner. Flavoring, preservative, dispersing and coloring agents may also be present.
Capsules can be prepared by preparing a powder mixture as described above and filling into shaped gelatin capsule shells. Glidants and lubricants, such as highly disperse silicic acid, talc, magnesium stearate, calcium stearate or polyethylene glycol in solid form, can be added to the powder mixture before the filling operation. Disintegrating or solubilizing agents (e.g., agar-agar, calcium carbonate or sodium carbonate) may also be added to enhance the availability of the drug after ingestion of the capsule.
In addition, if desired or necessary, suitable binders, lubricants and disintegrants and also dyes can be added to the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, sweeteners made from corn, natural and synthetic gums (such as 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, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. Tablets may be prepared, for example, by preparing a powder mixture, granulating or dry-compressing the mixture, adding a lubricant and disintegrant, and compressing the entire mixture into a tablet. Powder mixtures may be prepared by mixing the compound comminuted in a suitable manner with the above-mentioned diluents or bases and optionally with binders such as carboxymethylcellulose, alginates, gelatin or polyvinylpyrrolidone, dissolution retarders such as paraffin, absorption accelerators such as quaternary salts and/or absorbents such as bentonite, kaolin or dicalcium phosphate. The powder mixture may be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and sieving. As an alternative to granulation, the powder mixture may be passed through a tablet press to obtain a non-uniformly shaped mass which is broken up to form granules. The granules may be lubricated by the addition of stearic acid, a stearate salt, talc or mineral oil to prevent sticking to the tablet mould. The lubricated mixture is then compressed into tablets. It is also possible to combine the compounds of the present invention with free flowing inert excipients and then compress directly into tablets without a granulation or dry compression step. There may be a transparent or opaque protective layer consisting of a shellac barrier layer, a sugar or polymer material layer and a glossy layer of wax. Dyes may be added to these coatings to enable differentiation between different dosage units.
Oral liquids such as solutions, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in an aqueous solution with a suitable flavoring agent, while elixirs can be prepared using a non-toxic alcoholic medium. Suspensions may be prepared by dispersing the compound in a non-toxic vehicle. Solubilizing agents and emulsifiers such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers, preservatives, flavoring additives such as 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. The formulations may also be prepared in a form in which the release is extended or delayed, for example by coating or embedding the particulate material with a polymer, wax or the like.
The compounds of formula (I) and salts, solvates and physiologically functional derivatives thereof may also be administered in the form of liposomal delivery systems such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
The compounds of formula (I) and salts, solvates and physiologically functional derivatives thereof may also be delivered using a monoclonal antibody as a separate carrier, wherein the compound molecule is coupled to the monoclonal antibody. The compounds may also be coupled to soluble polymers as targeted drug carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenol, or polyoxyethylene polylysine, substituted with palmitoyl groups. The compounds may also be coupled to a class of biodegradable polymers suitable for achieving controlled release of a drug, such as polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, and cross-linked or amphiphilic block copolymer hydrogels.
Pharmaceutical formulations adapted for transdermal administration may be administered as a stand-alone plaster in intimate contact with the epidermis of the recipient for an extended period of time. Thus, for example, the active ingredient may be delivered from a plaster by iontophoresis, as described in general terms in Pharmaceutical Research, 3(6), 318 (1986).
Pharmaceutical compounds suitable for topical administration can be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.
For the treatment of the eye or other external tissues (e.g. oral cavity and skin), the formulations are preferably applied in the form of a topical ointment or cream. In the case of formulating an ointment, the active ingredient may be applied with a paraffinic or water-miscible cream base. Alternatively, the active ingredient may be formulated as a cream in an oil-in-water cream base or a water-in-oil base.
Pharmaceutical formulations suitable for topical application to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, particularly an aqueous solvent.
Pharmaceutical formulations adapted for topical application in the mouth include lozenges, pastilles and mouthwashes.
Pharmaceutical preparations suitable for rectal administration may be administered in the form of suppositories or enemas.
Pharmaceutical preparations suitable for nasal administration in which the carrier material is a solid comprise a coarse powder having a particle size of, for example, 20 to 500 microns which can be administered by sniffing, i.e. rapid inhalation from a powder-containing container near the nose via the nasal passage. Suitable formulations for administration as nasal sprays or nasal drops containing a liquid as carrier material comprise solutions of the active ingredient in water or oil.
Pharmaceutical formulations adapted for administration by inhalation comprise a fine particulate powder or mist which may be generated by various types of aerosol-containing pressurised dispensers, nebulisers or insufflators.
Pharmaceutical formulations adapted for vaginal administration may be administered in the form of pessaries, tampons, creams, gels, pastes, foams or spray formulations.
Pharmaceutical formulations suitable for parenteral administration include: aqueous and non-aqueous sterile injection solutions containing antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the recipient to be treated; aqueous and non-aqueous sterile suspensions, which may contain suspending media and thickening agents. The formulations may be administered in single-or multi-dose containers, for example sealed ampoules and vials, and stored in a freeze-dried (lyophilized) condition, so that the addition of a sterile carrier liquid, for example water for injections, is required only immediately prior to use. Injection solutions and suspensions prepared according to the prescription can be prepared from sterile powders, granules, and tablets.
It goes without saying that, in addition to the components specifically mentioned above, the formulations may also comprise other substances usable in the art for this particular type of formulation; thus, for example, formulations suitable for oral administration may contain flavouring agents.
The therapeutically effective amount of a compound of formula (I) will depend on a number 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 discretion of the attendant physician or veterinarian. However, an effective amount of a compound of the invention for use in the treatment of tumor growth, e.g., colon or breast cancer, is generally from 0.1 to 100mg/kg of recipient (mammalian) body weight per day, and in particular is typically from 1 to 10mg/kg of body weight per day. Thus, for an adult mammal weighing 70kg, the actual amount per day will typically be 70 to 700mg, wherein the amount may be administered as a single dose per day or typically in a plurality of divided doses per day (e.g. two, three, four, five or six divided doses) such that the total daily dose is the same. An effective amount of a salt or solvate or physiologically functional derivative thereof may be determined as a fraction of the effective amount of the compound of the invention per se. It is believed that similar dosages are suitable for the treatment of the other conditions mentioned above.
The invention also relates to a medicament comprising at least one compound of the formula (I) and/or pharmaceutically acceptable salts and stereoisomers thereof, including mixtures thereof in all ratios, and also at least one further pharmaceutically active ingredient.
The invention also relates to a kit (kit) consisting of the following individual packages:
(a) an effective amount of a compound of formula (I) and/or pharmaceutically acceptable salts and stereoisomers thereof (including mixtures thereof in all ratios);
and
(b) an effective amount of other pharmaceutically active ingredients.
The kit comprises a suitable container such as a box, individual bottle, pouch or ampoule. The kit may comprise, for example, separate ampoules, each containing an effective amount of a compound of formula (I) and/or pharmaceutically acceptable derivatives, solvates and stereoisomers thereof, including mixtures thereof in all ratios, and an effective amount of the other pharmaceutically active ingredient in dissolved or freeze-dried form.
Applications of
The compounds of the present invention are suitable as pharmaceutical active ingredients for mammals, especially humans, for the treatment of tyrosine kinase induced diseases. These diseases include tumor cell proliferation, pathological neovascularization (or angiogenesis) that promotes the growth of solid tumors, ocular neovascularization (diabetic retinopathy, age-induced macular degeneration, etc.), and inflammation (psoriasis, rheumatoid arthritis, etc.).
The invention comprises the use of a compound of formula (I) and/or physiologically acceptable salts and solvates thereof in the manufacture of a medicament for the treatment or prevention of cancer. Preferred cancers to be treated are cancers derived from brain, genitourinary tract, lymphatic system, stomach, larynx and lung cancers. Another group of preferred cancer forms are monocytic leukemia, lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma and breast cancer.
Also included is the use of a compound of the invention as claimed in claim 1 and/or physiologically acceptable salts and solvates thereof for the manufacture of a medicament for the treatment or prevention of diseases in which angiogenesis is involved.
Such diseases in which angiogenesis is involved are ocular diseases such as retinal vascularization, diabetic retinopathy, age-induced macular degeneration and the like.
The use of a compound of formula (I) and/or physiologically acceptable salts and solvates thereof in the manufacture of a medicament for the treatment or prevention of inflammatory diseases is also within the scope of the present invention. Examples of such inflammatory diseases include rheumatoid arthritis, psoriasis, contact dermatitis, delayed type hypersensitivity reactions, and the like.
Also included is the use of a compound of formula (I) and/or physiologically acceptable salts and solvates thereof, for the manufacture of a medicament for the treatment or prevention of a tyrosine kinase-induced disease or a tyrosine kinase-induced disorder in a mammal, wherein for the method a therapeutically effective amount of a compound of the invention is administered to a diseased mammal in need of such treatment. The amount of treatment will vary depending on the particular disease and can be determined by one skilled in the art without undue experimentation.
The invention also comprises the use of a compound of formula (I) and/or physiologically acceptable salts and solvates thereof in the manufacture of a medicament for the treatment or prevention of retinal vascularization.
Methods for treating or preventing ocular diseases such as diabetic retinopathy and age-induced macular degeneration are also part of the invention. The use for the treatment or prevention of inflammatory diseases such as rheumatoid arthritis, psoriasis, contact dermatitis and delayed hypersensitivity and the use for the treatment or prevention of bone diseases derived from osteosarcoma, osteoarthritis and rickets also belong to the scope of the present invention.
The expression "tyrosine kinase-induced disease or condition" refers to a pathological condition which is dependent on the activity of one or more tyrosine kinases. Tyrosine kinases are involved directly or indirectly in the signal transduction pathways of a variety of cellular activities, including proliferation, adhesion and migration, and differentiation. Diseases associated with tyrosine kinase activity include tumor cell proliferation, pathological neovascularization that promotes the growth of solid tumors, ocular neovascularization (diabetic retinopathy, age-induced macular degeneration, etc.), and inflammation (psoriasis, rheumatoid arthritis, etc.).
The compounds of formula (I) may be administered to a patient to treat cancer, particularly rapidly growing tumors.
The invention therefore relates to the use of compounds of formula (I) and their pharmaceutically acceptable salts and stereoisomers, including mixtures thereof in all ratios, in the manufacture of a medicament for the treatment of diseases in which inhibition, modulation and/or modulation of kinase signal transduction plays a role.
Preferred herein is Met kinase.
Preferably the use of a compound of formula (I) and pharmaceutically acceptable salts and stereoisomers thereof, including mixtures thereof in all ratios, in the manufacture of a medicament for the treatment of diseases in which inhibition of tyrosine kinases by the compounds of claim 1 has an effect.
Particularly preferred is the use for the preparation of a medicament for the treatment of diseases in which inhibition of Met kinase by a compound according to claim 1 has an effect.
Especially preferred is the use for the treatment of a disease, wherein the disease is a solid tumor.
The solid tumor is preferably selected from tumors of the lung, squamous epithelium, bladder, stomach, kidney, head and neck, esophagus, cervix, thyroid, intestine, liver, brain, prostate, genito-urinary tract, lymphatic system, stomach and/or larynx.
The solid tumor is also preferably selected from lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma, colon carcinoma and breast carcinoma.
Also preferred is the use for the treatment of tumours of the blood and immune system, preferably for the treatment of tumours selected from acute myeloid leukaemia, chronic myeloid leukaemia, acute lymphatic leukaemia and/or chronic lymphatic leukaemia.
The disclosed compounds of formula (I) may be used in combination with other known therapeutic ingredients, including anticancer agents. The term "anti-cancer agent" as used herein refers to any ingredient that is administered to a cancer patient for the purpose of treating cancer.
The anti-cancer treatments defined herein may be applied as monotherapy or may include conventional surgery or radiotherapy or chemotherapy in addition to the compounds of the invention. Such chemotherapy may include one or more of the following classes of antineoplastic agents:
(i) antiproliferative/antineoplastic/DNA-damaging agents and combinations thereof for medical oncology, such as alkylating agents (e.g., cisplatin, carboplatin, cyclophosphamide, mechlorethamine, melphalan, chlorambucil, busulfan, and nitrosoureas); antimetabolites (e.g., antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytarabine, hydroxyurea, and gemcitabine); antitumor antibiotics (e.g., anthracyclines such as doxorubicin, bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin); antimitotic agents (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxanes such as taxol and taxotere); topoisomerase inhibitors (e.g. epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, irinotecan and camptothecin) and cell differentiating agents (e.g. all-trans retinoic acid, 13-cis-retinoic acid and fenretinide);
(ii) cytostatics, such as antiestrogens (e.g., tamoxifen, toremifene, raloxifene, droloxifene, and iodoxyfene), estrogen receptor downregulators (e.g., fulvestrant), antiandrogens (e.g., bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or LHRH agonists (e.g., goserelin, leuprolide, and buserelin), progestins (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, vorozole, and exemestane), and 5 α -reductase inhibitors such as finasteride;
(ii) components that inhibit cancer cell invasion (e.g., metalloproteinase inhibitors such as marimastat and urokinase-plasminogen activator receptor function inhibitors);
(iv) inhibitors of growth factor function, e.g., such inhibitors include growth factor antibodies, growth factor receptor antibodies (e.g., anti-erbb 2-antibody trastuzumab [ Herceptin ]TM]And the anti-erbb 1-antibody cetuximab [ C225]) Farnesyl transferase inhibitors, tyrosine kinase inhibitors and serine/threonine kinase inhibitors, e.g. inhibitors of the epidermal growth factor family (e.g. EGFR family tyrosine kinase inhibitors, such asN- (3-chloro-4-fluorophenyl) -7-methoxy-6- (3-morpholinopropoxy) quinazolin-4-amine (gefitinib, AZD1839),N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) quinazolin-4-amine (erlotinib, OSI-774) and 6-acrylamido-N- (3-chloro-4-fluorophenyl) -7- (3-morpholinopropoxy) quinazolin-4-amine (Cl 1033)); inhibitors such as the platelet derived growth factor family and inhibitors such as the hepatocyte growth factor family;
(v) anti-angiogenic agents, such as those that inhibit the action of vascular endothelial growth factor (e.g., the anti-vascular endothelial growth factor antibody bevacizumab [ Avastin ]TMSuch as those disclosed in published international patent applications WO 97/22596, WO 97/30035. Those of WO 97/32856 and WO 98/13354) and compounds that act by other mechanisms (e.g., linoxamine, integrin- α v β 3 function inhibitors and angiostatin);
(vi) vascular damaging agents, such as combretastatin a4 and compounds disclosed in international patent applications WO 99/02166, WO00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213;
(vii) antisense therapies, e.g., those directed against the above targets, such as ISIS 2503, anti-Ras antisense;
(viii) gene therapy methods, including, for example, methods of replacing abnormal genes such as abnormal p53 or abnormal BRCA1 or BRCA 2; GDEPT (gene-directed enzyme prodrug therapy) approaches such as those using cytosine deaminase, thymidine kinase, or bacterial nitroreductase; and methods of increasing the patient's tolerance to chemotherapy or radiation therapy, such as multi-drug resistance gene therapy; and
(ix) immunization methods, including, for example, ex vivo and in vivo methods of increasing the immunogenicity of patient tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte macrophage colony stimulating factor; methods of reducing T-cell anergy; methods using transfected immune cells such as cytokine-transfected dendritic cells; methods of using cytokine-transfected tumor cell lines, and methods of using anti-atopic antibodies.
Preferably the drugs in table 1 below (but not exclusively) are used in combination with a compound of formula (I).
Such combination therapy can be achieved by means of simultaneous, sequential or separate administration of the individual components of the treatment. This type of combination uses the compounds of the present invention.
Measurement method
The compounds of formula (I) described in the examples were tested by the assays described below and found to have kinase inhibitory activity. Other assays are known In the literature and can be readily performed by those skilled In the art (see, e.g., Dhanabal et al, Cancer Res.59: 189-197; Xin et al, J.biol.chem.274: 9116-9121; Sheu et al, Anticancer Res.18: 4435-4441; Ausprunk et al, Dev.biol.38: 237-248; Gimbron et al, J.Natl.cancer Inst.52: 413-427; Nicosia et al, In Vitro 18: 538-549).
Determination of Met kinase Activity
According to the manufacturer's data (Met, activity, upstate, cat # 14-526), Met kinase was expressed with the aim of producing proteins in insect cells (St 21; Spodoptera frugiperda), followed by affinity chromatography purification in the form of an "N-terminal 6 His-tagged" recombinant human protein in a baculovirus expression vector.
Kinase activity can be measured using a variety of available assay systems. Radiolabeled ATP is used in scintillation proximity assays (Sorg et al, J.of. Biomolecular Screening, 2002, 7, 11-19), flash plate methods, or filter binding assays (II)32p-ATP，33p-ATP) to measure the radioactive phosphorylation of proteins or peptides as substrates. In the presence of the inhibitory compound, a reduced or no radioactive signal is detected. In addition, homogeneous time-resolved fluorescence resonance energy transfer (HTR-FRET) and Fluorescence Polarization (FP) techniques can also be used as assay methods (Sills et al, J.of Biomolecular Screening, 2002, 191-214).
Other non-radioactive ELISA assays use specific phospho-antibodies (phospho-AB). Phospho-antibodies bind only phosphorylated substrates. This binding can be detected using a peroxidase-conjugated secondary antibody based on chemiluminescence (Ross et al, 2002, biochem.j.).
Flash plate method (Met kinase)
The test plate used was a 96-well Flashplate from Perkin EimerRMicroplate (catalog number SMP 200). The components of the kinase reaction described below were pipetted into the assay plate. Met kinase and substrate poly Ala-Glu-Lys-Tyr (pAGLT, 6: 2: 5: 1) are reacted with radiolabeled33p-ATP was incubated together in a total volume of 100. mu.l for 3 hours at room temperature in the presence and absence of the experiment. The reaction was stopped using 150. mu.l of 60mM EDTA solution. After a further incubation for 30 minutes at room temperature, the supernatant was filtered off with suction and the wells were washed three times with 200. mu.l of 0.9% NaCl solution each time. Bound radioactivity was determined by scintillation assay (topcount nxt, Perkin Eimer).
The full value used is the kinase response without inhibitor. This should be between about 6000 and 9000 cpm. The pharmacological zero value used was staurosporine at a final concentration of 0.1 mM. Inhibition values were determined using the RS1_ MTS program (IC 50).
Kinase reaction conditions in each well:
30 μ l assay buffer
10 μ l of test substance in assay buffer containing 10% DMSO
10 μ l ATP (final concentration 1 μ M non-radioactive, 0.35 μ Ci)33p-ATP)
50 μ l of Met kinase/substrate mixture in assay buffer;
(10ng enzyme/well, 50ng pAGLT/well)
The solutions used were:
-assay buffer:
50mM HEPES
3mM magnesium chloride
3 μ M sodium orthovanadate
3mM manganese (II) chloride
1mM Dithiothreitol (DTT)
pH 7.5 (set using sodium hydroxide)
-a stop solution:
60mM Titriplex Ⅲ(EDTA)
-33p-ATP：Perkin-Elmer；
-Met kinase: upstate, catalog number 14-526, stock 1 μ g/10 μ l; specific activity 954U/mg;
-poly-Ala-Glu-Lys-Tyr, 6: 2: 5: 1: sigma catalog number P1152
In vivo experiment (fig. 1/1)
Experimental methods: female Balb/C mice (breeder: Charles River VVwiga) were 5 weeks old at arrival. The animals were acclimated to the experimental breeding environment for 7 days. 400 million TPR-Met/NIH3T3 cells in 100. mu.l PBS (without Ca + + and Mg + +) were injected subcutaneously at the pelvic site of each mouse. After 5 days, the mixture is preparedThe animals were randomly divided into 3 groups, and the average tumor volume of 9 mice per group was 110. mu.l (range: 55-165). The control group was given 100. mu.l of vehicle (0.25% methylcellulose/100 mM acetate buffer, pH 5.5) daily, and the treatment group was given 200mg/kg of "A56" or "A91" dissolved in vehicle (volume likewise 100. mu.l/animal) daily, both via gastric tube. After 9 days, the mean volume of the control group was 1530. mu.l, and the experiment was stopped.
Determination of tumor volume: the length (L) and width (B) were measured with a vernier caliper and the tumor volume was calculated using the formula L.times.BxB/2.
Feeding conditions: animals were fed with commercial mouse chow (Sniff) 4 or 5 per cage.
The compounds "A18" and "A22" have significant antitumor effects.
In this context, all temperatures are expressed in degrees Celsius. In the following examples, "conventional treatment" means: if necessary, adding water; if necessary, the pH is adjusted to 2-10, depending on the constitution of the end product, the mixture is extracted with ethyl acetate or dichloromethane, the phases are separated, the organic phase is dried over sodium sulfate, evaporated, and the residue is purified by chromatography on silica gel and/or by crystallization. Rf value on silica gel; eluent: ethyl acetate/methanol 9: 1.
mass Spectrum (MS): EI (Electron impact ionization) M+
FAB (Rapid atom bombardment) (M + H)+
ESI (electrospray ionization) (M + H)+
APCI-MS (atmospheric pressure chemical ionization-mass spectrometry) (M + H)+。
Mass Spectrum (MS): EI (Electron impact ionization) M+
FAB (Rapid atom bombardment) (M + H)+
ESI (electrospray ionization) (M + H)+
APCI-MS (atmospheric pressure chemical ionization-mass spectrometry) (M + H)+。
HPLC method:
HPLC/MS analysis
On a 3 μ Silica-Rod column, with a gradient elution of 20-100% water/acetonitrile/0.01% trifluoroacetic acid for 210 seconds, at a flow rate of 2.2ml/min, at 220 nm.
HPLC analysis (method A)
Column: chromolith RP18e 100 x 3mm
Flow rate: 2ml/min
Solvent A: h2O + 0.1% trifluoroacetic acid
Solvent B: acetonitrile + 0.1% trifluoroacetic acid
Gradient elution for 5 min
0-4 minutes: 99: 1- > 1: 99
4-5 minutes: 1: 99-1: 99
HPLC analysis (method B)
Column: chromolith RP18e 100 x 3mm
Flow rate: 4ml/min
Solvent A: h2O+0.05％HCOOH
Solvent B: acetonitrile + 10% solvent A
Gradient elution for 8 min
0-1 minute: 99: 1- > 99: 1
1-7 minutes: 99: 1-1: 99
7-8 minutes: 1: 99- > 1: 99
Retention time Rt is in minutes.
Example 1
The preparation of 2- [3- (5-bromopyrimidin-2-yl) benzyl ] -6-cyclopropyl-2H-pyridazin-3-one ("a 1") and 6-cyclopropyl-2- (3- {5- [ 1- (2-pyrrolidin-1-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one ("a 2") was carried out according to the following scheme:
1.1 16.6g (180mmol) of glyoxylic acid monohydrate and 50ml (535mmol) of cyclopropylmethyl ketone are heated at 120 ℃ and stirred for 2 hours. The reaction mixture was cooled to 40 ℃ and 70ml of water and 14ml of 32% aqueous ammonia solution were added. The mixture was extracted three times with dichloromethane. 8.7ml (179mmol) of hydrazine hydroxide are added to the aqueous phase and the mixture is heated at reflux for 1 hour. The reaction mixture was cooled to room temperature. The precipitate formed is filtered off with suction, washed with brine and dried in vacuo: 6-cyclopropyl-2H-pyridazin-3-one as colorless crystals; ESI 137.
1.2A 1.5l aqueous solution of 318g (3.00mmol) of sodium carbonate is added to a 1.5l toluene solution of 427g (1.50mol) of 5-bromo-2-iodopyrimidine kept under nitrogen. The mixture was heated to 80 ℃ and a solution of 34.7g (30mmol) tetrakis (triphenylphosphine) palladium and 228g (1.50mol)3- (hydroxymethyl) benzeneboronic acid in 3l of ethanol was added. The reaction mixture was then heated at reflux for 18 hours. The reaction mixture was cooled to room temperature, filtered and partitioned between water and tert-butyl methyl ether. The organic phase was dried over sodium sulfate and concentrated to a volume of 1 l. The precipitate formed is filtered off with suction and washed with toluene: [3- (5-bromopyrimidin-2-yl) phenyl ] methanol as a colorless crystal; ESI 265, 267.
1.3A solution of 2.23g (8.41mmol) of [3- (5-bromopyrimidin-2-yl) phenyl ] methanol, 1.49g (10.9mmol) of 6-cyclopropyl-2H-pyridazin-3-one and 3.34g (12.6mmol) of triphenylphosphine in 80ml of THF, kept under nitrogen, was cooled in an ice bath and 2.61ml (12.6mmol) of diisopropyl azodicarboxylate (DIAD) was slowly added dropwise. The reaction mixture was stirred at room temperature for 2 hours and evaporated. The residue was chromatographed on a column of silica gel, eluting with dichloromethane/methanol. The product-containing fractions were combined, evaporated and recrystallized from isopropanol: 2- [3- (5-bromopyrimidin-2-yl) benzyl ] -6-cyclopropyl-2H-pyridazin-3-one ("Al") as a colorless crystal; ESI 383, 385;
1H-NMR(DMSO-d6)：δ【ppm]0.80(m，2H)，0.93(m，2H)，1.96(m，1H)，5.(s，2H)，6.91(d，J＝9.5Hz，1H)，7.29(d，J＝9.5Hz，1H)，7.46(dt，J1＝7.5Hz，J2＝1Hz，1H)，7.51(t，J＝7.5Hz，1H)，8.27(dt，J1＝7.5Hz，J2＝1Hz，1H)，8.30(t，J＝1Hz，1H)，9.07(s，2H)。
1.4 15.0g (86.7mmol) of N- (2-chloroethyl) pyrrolidine hydrochloride and 42.4g (130mmol) of cesium carbonate are added to a solution of 8.58g (43.3mmol) of pyrazole-4-boronic acid pinacol ester in 86ml of acetonitrile and the resulting suspension is stirred at room temperature for 18 hours. The reaction mixture was filtered and the residue was washed with acetonitrile. The filtrate was evaporated and the residue partitioned between saturated sodium chloride solution and ethyl acetate. The organic phase was dried over sodium sulfate and evaporated: 10.9g of 1- (2-pyrrolidin-1-ylethyl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole as a yellow oil; ESI 292.
1.5 to a solution of 394mg (1.00mmol)2- [3- (5-bromopyrimidin-2-yl) benzyl ] -6-cyclopropyl-2H-pyridazin-3-one and 337mg (1.10mmol)1- (2-pyrrolidin-1-ylethyl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole maintained under nitrogen, 425mg (2.0mmol) tripotassium phosphate trihydrate and 56.2mg (0.08mmol) bis (triphenylphosphine) palladium chloride are added and the mixture is stirred at 80 ℃ for 18 hours. The reaction mixture was filtered through celite, washing with dichloromethane. The filtrate was evaporated and the residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: 6-cyclopropyl-2- (3- {5- [ 1- (2-pyrrolidin-1-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one ("a 2") as a colorless oil; ESI 468;
1H-NMR(DMSO-d6)：δ【ppm】0.82(rn，2H)，0.95(m，2H)，1.68(m，4H)，1.(m，1H)，2.51(m，4H)，2.88(t，J＝6.8Hz，2H)，4.28(t，J＝6.8Hz，2H)，5.(s，2H)，6.93(d，J＝9.5Hz，1H)，7.31(d，J＝9.5Hz，1H)，7.42(d，J＝7.5Hz，1H)，7.50(t，J＝7.5Hz，1H)，8.11(s，1H)，8.31(d，J＝7.5Hz，1H)，8.33(bs，1H)，8.45(s，1H)，9.14(s，2H)。
the following compounds were obtained according to a similar method:
6-cyclopropyl-2- (3- {5- [ 1- (2-morpholin-4-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one
6-cyclobutyl-2- (3- {5- [ 1- (2-pyrrolidin-1-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-yl } benzyl) -2H-pyridazin-3-one
1H-NMR (d 6-DMSO): δ [ ppm ] 1.67(m, 4H), 1.83(m, 1H), 1.97(m, 1H), 2.21(m, 4H), 2.49(m, 4H), 2.87(t, J ═ 6.8Hz, 2H), 3.50 (quintuple, J ═ 8.5Hz, 1H), 4.26(t, J ═ 6.8Hz, 2H), 5.32(s, 2H), 6.96(d, J ═ 9.5Hz, 1H), 7.43(d, J ═ 7.5Hz, 1H), 7.45(d, J ═ 9.5Hz, 1H), 7.50(t, J ═ 7.5Hz, 1H), 8.10(s, 1H), 8.30(d, J ═ 7.5, 1H), 8.36.36 (bs), 8.12H, 8.7 (s, 2H), 8.6.6.5 Hz, 1H).
Example 2
The preparation of 6-cyclopropyl-2- [3- (5-methylpyrimidin-2-yl) benzyl ] -2H-pyridazin-3-one ("a 4") was carried out analogously according to the following scheme:
2.1 to a suspension of 2.41g (10.0mmol) of methyl 3-formamidinibenzoate acetate in 40ml of methanol, 1.31ml (11.0mmol) of 3-ethoxymethacrolein and 2.04ml (11.0mmol) of 30% sodium ethoxide in methanol are added and the resulting solution is stirred at 50 ℃ for 18 hours. The reaction mixture was evaporated in vacuo and water was added. The precipitate formed is filtered off with suction, washed with water and dried in vacuo: 3- (5-methylpyrimidin-2-yl) benzoic acid methyl ester as colorless crystals; the ESI 229.
2.2 600mg (5.41mmol) of calcium chloride powder are added to a suspension of 400mg (10.6mmol) of sodium borohydride in 20ml of THF, and the mixture is stirred at room temperature for 1.5 hours. To this suspension was added dropwise, while stirring, a solution of 751mg (3.29mmol) of methyl 3- (5-methylpyrimidin-2-yl) benzoate in 10ml of THF, and the mixture was stirred at room temperature for 18 hours. To the reaction mixture was added 10ml of 1N NaOH, water and methylene chloride, which was then filtered. The organic phase of the filtrate was separated, dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: [3- (5-methylpyrimidin-2-yl) phenyl ] methanol as a colorless solid; ESI 201.
2.3 mu.l (0.75rnmol) of diisopropyl azodicarboxylate are added dropwise to a solution of 68.1mg (0.50mmol) of 6-cyclopropyl-2H-pyridazin-3-one, 100mg (0.50mmol) of [3- (5-methylpyrimidin-2-yl) phenyl ] methanol and 197mg (0.75mmol) of triphenylphosphine in 3ml of THF and the solution obtained is stirred at room temperature for 18H. The reaction mixture was evaporated in vacuo and the residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: 6-cyclopropyl-2- [3- (5-methylpyrimidin-2-yl) benzyl ] -2H-pyridazin-3-one ("a 4") as a colorless solid; ESI 319.
Example 3
Preparation of 6-cyclopropyl-2- {3- [5- (1-piperidin-4-yl-1H-pyrazol-4-yl) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("a 5") was carried out according to the following scheme:
3.1 637mg (3.0mrnol) of tripotassium phosphate trihydrate and 84.2mg (0.12mmol) of bis (triphenylphosphine) palladium chloride are added to a solution of 599mg (1.50mmol) of 2- [3- (5-bromopyrimidin-2-yl) benzyl ] -6-cyclopropyl-2H-pyridazin-3-one and 623mg (1.65mmol) of 4- [ 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl ] -piperidine-1-carboxylic acid tert-butyl ester (prepared according to the method described in WO 2007/066187) in 15ml of 1, 2-dimethoxyethane which is kept under nitrogen, and the mixture is stirred at 80 ℃ for 18 hours. The reaction mixture was filtered through celite, washing with dichloromethane. The filtrate was washed with water. The organic phase was dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: 4- (4- {2- [3- (3-cyclopropyl-6-oxo-6H-pyridazin-1-ylmethyl) phenyl ] -pyrimidin-5-yl } pyrazol-1-yl) piperidine-1-carboxylic acid tert-butyl ester as a pale yellow high viscosity oil; ESI 554.
3.2 to a suspension of 692mg (1.24mmol) of tert-butyl 4- (4- {2- [3- (3-cyclopropyl-6-oxo-6H-pyridazin-1-ylmethyl) phenyl ] pyrimidin-5-yl } pyrazol-1-yl) piperidine-1-carboxylate in 5ml of 4N HCI in dioxane was added 2ml of methanol and the mixture was heated at 80 ℃ for 5 minutes. The reaction mixture was evaporated and water, dichloromethane and saturated sodium bicarbonate solution were added sequentially. The organic phase was separated, dried over sodium sulfate and evaporated. The residue was recrystallized from tert-butyl methyl ether: 6-cyclopropyl-2- {3- [5- (1-piperidin-4-yl-1H-pyrazol-4-yl) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("a 5") as colorless crystals; ESI 454;
1H-NMR(DMSO-ds)：δ【ppm】0.81(m，2H)，0.94(m，2H)，1.80(m，2H)，1.(m，3H)，2.60(m，2H)，3.05(m，2H)，4.23(m，1H)，5.27(s，2H)，6.92(d，J＝9.5Hz，1H)，7.30(d，J＝9.5Hz，1H)，7.41(d，J＝7.5Hz，1H)，7.49(t，J＝7.5Hz，1H)，8.10(s，1H)，8.30(d，J＝7.5Hz，1H)，8.33(bs，1H)，8.48(s，1H)，9.14(s，2H)。
compound (I)
6-methyl-2- {3- [5- (1-piperidin-4-yl-1H-pyrazol-4-yl) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one obtained according to an analogous method:
example 4
The preparation of 6-cyclopropyl-2- [3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzyl ] 2H-pyridazin-3-one ("a 7") was carried out analogously according to the following scheme:
169mg (1.0mmol) of cesium carbonate are added to a solution of 68.1mg (0.50mol) of 6-cyclopropyl-2H-pyridazin-3-one and 126.5mg (0.50mmol) of 3- (3-bromomethylphenyl) -5-methyl-1, 2, 4-oxadiazole (prepared according to the method described in W.W.K.R.Mederski et al, Tetrahedron 55, 1999, 12757-. To the reaction mixture was added water and tert-butyl methyl ether. The organic phase was separated, dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: 6-cyclopropyl-2- [3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzyl ] -2H-pyridazin-3-one ("A7") as a colourless crystal; ESI 309.
Example 5
The preparation of 6-cyclopropyl-2- {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("A8") was carried out analogously according to the following scheme:
5.1 to a suspension of 500g (3.40mol) of 3-cyanobenzoic acid in 8l of methanol maintained at 30 ℃ 1382g (10.0mol) of potassium carbonate are added portionwise with stirring. Subsequently 695g (10.0mol) of hydroxylammonium chloride are added in small portions each, maintaining the internal temperature at 40-45 ℃. The reaction mixture was then heated at boiling for 15 hours. The reaction mixture was evaporated in vacuo, dissolved in water and acidified with 37% aqueous hydrochloric acid. The precipitate formed is filtered off with suction, washed with water and dried in vacuo: 3- (N-hydroxycarbamimidoyl) -benzoic acid as colorless crystals; ESI 181.
5.2A mixture of 614g (3.41mol) of 3- (N-hydroxycarbamimidoyl) benzoic acid, 756ml (8.0mol) of acetic anhydride and 2l of acetic acid is heated at 118 ℃ for 14 hours. The reaction mixture was cooled to 6 ℃ and filtered with suction. The residue is dissolved in 2l of water, filtered off with suction and washed thoroughly with water. The residue was recrystallized in ethanol/water: 3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzoic acid as colorless crystals; m.p.225 ℃; ESI 205.
5.3 7.83ml (147mmol) of concentrated sulfuric acid are added to a suspension of 30.0g (147mmol) of 3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzoic acid in 150ml of methanol and the mixture is heated at the boil for 18 hours. The reaction mixture was cooled in an ice bath, water was added, the product was filtered off with suction and washed thoroughly with water: 3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzoic acid methyl ester as colorless crystals; the ESI 219.
54 150ml of acetic acid, 150ml of water and 50g of water-moist Raney nickel are added to a solution of 327g (1.47mol) of methyl 3- (5-methyl-1, 2, 4-oxadiazol-3-yl) benzoate in 3l of methanol and the mixture is hydrogenated at room temperature and atmospheric pressure for 18 hours. The catalyst is filtered off and the filtrate is evaporated. The residue is dissolved in tert-butyl methyl ether, heated to boiling and filtered off with suction. The residue was dried in vacuo: 3-methoxycarbonylbenzamidine (benzamidinium) acetate as a colorless crystal; ESI 179.
5.5.21 freshly prepared 1.5M sodium methoxide solution was added dropwise with stirring to a suspension of 259g (1.09mol) of 3-methoxycarbonylbenzamidine acetate and 528g (1.08mol) of ({ 2-dimethylamino-1- [ dimethylammonio (immonoi) methyl ] vinylamino } methylene) dimethylammonium dihexafluorophosphate (prepared according to the method described in C.B.Dousson et al, Synthesis 2005, 1817) in 1l of methanol. The reaction mixture was then warmed to 60 ℃ over 40 minutes, and held at this temperature for 30 minutes. The reaction mixture was then cooled to room temperature, diluted with 101 dichloromethane and washed three times with 5l of water. The organic phase was dried over sodium sulfate and evaporated. The residue was recrystallized from ethyl acetate: 3- [5- (dimethylamino methyleneamino) pyrimidin-2-yl ] benzoic acid methyl ester as beige crystals; m.p.140 ℃; ESI 285.
5.6 to a 1.3l aqueous suspension of 103.5g (364mmol) of methyl 3- [5- (dimethylaminomethyleneamino) pyrimidin-2-yl ] benzoate 160ml (2.88mol) of concentrated sulfuric acid are added and the mixture is heated at the boil for 4 hours. The reaction mixture was cooled to room temperature, diluted with water and filtered with suction. The residue was washed with water and dried in vacuo: 3- (5-hydroxypyrimidin-2-yl) benzoic acid as light brown crystals; ESI 217.
5.7 to a suspension of 88.0g (366mmol) of 3- (5-hydroxypyrimidin-2-yl) benzoic acid in 1.4l of methanol was added 32.7ml (445mmol) of thionyl chloride, and the mixture was heated at 80 ℃ for 2 hours. 20ml (276mmol) of thionyl chloride are added, followed by another 10ml (138mmol) of thionyl chloride. After each addition, the reaction mixture was stirred at 80 ℃ for 2 hours. The reaction mixture was concentrated in vacuo to a volume of about 300ml, the precipitate formed was filtered and dried in vacuo: 3- (5-hydroxypyrimidin-2-yl) benzoic acid methyl ester as light brown crystals; ESI 231.
5.8A solution of 6.1g (26.5mmol) of methyl (5-hydroxypyrimidin-2-yl) benzoate, 10.5g (39.8mmol) of triphenylphosphine and 4.76ml (39.8mmol) of 3- (dimethylamino) -1-propanol in 200ml of THF, kept under nitrogen, is cooled in an ice bath and 8.21ml (39.8mmol) of diisopropyl azodicarboxylate is slowly added dropwise with stirring. After stirring the reaction mixture at room temperature for 2 hours, it was evaporated in vacuo. The residue was partitioned between dichloromethane and saturated aqueous potassium hydrogen sulfate. The aqueous phase was separated, the pH adjusted to 12 with saturated aqueous sodium hydroxide solution and extracted twice with dichloromethane. The organic phase was dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol: 3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzoic acid methyl ester as colorless crystals; ESI 316.
5.9 to a solution of 12.6g (40.0mmol) of methyl 3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzoate in 200ml of THF, kept under nitrogen, was added dropwise 200ml of a 1M solution of diisobutylaluminum hydride in THF with stirring. After the mixture was stirred at room temperature for 1 hour, 10ml of a saturated aqueous sodium sulfate solution was added dropwise. The precipitate formed is filtered off with suction and washed with dichloromethane. The filtrate was dried over sodium sulfate and evaporated. The residue was dissolved in a mixture of diethyl ether and petroleum ether. The precipitate formed is filtered off with suction, washed with petroleum ether and dried in vacuo: {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] phenyl } methanol as colorless crystals; m.p.95-97 deg.c; ESI 288.
5.10A suspension of 144mg (0.50mmol) {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] phenyl } methanol, 88.5mg (0.65mmol) 6-cyclopropyl-2H-pyridazin-3-one and 199mg (0.75mmol) triphenylphosphine in 1ml THF is cooled in an ice bath and 160mg (0.75mmol) diisopropyl azodicarboxylate is slowly added dropwise. After the mixture was stirred at room temperature for 1 hour, ethyl acetate and a 2N aqueous hydrochloric acid solution were added. The aqueous phase was separated and washed three times with ethyl acetate. The aqueous phase was adjusted to pH 14 with 2N sodium hydroxide solution and extracted twice with ethyl acetate. The organic phase was dried over sodium sulfate and evaporated. The residue was purified by preparative HPLC. The fractions containing the product were evaporated, dissolved in 0.5ml of 1N aqueous hydrochloric acid and a little water, freeze-dried: 6-cyclopropyl-2- {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("A8") hydrochloride as a colorless glass; ESI 406;
1H-NMR(DMSO-d6)：δ【ppm】0.80(m，2H)，0.93(m，2H)，1.96(m，1H)，2.(m，2H)，2.78(d，J＝5Hz，6H)，3.22(m，2H)，4.31(t，J＝6Hz，2H)，5.25(s，2H)，6.91(d，J＝9.5Hz，1H)，7.29(d，J＝9.5Hz，1H)，7.37(d，J＝7.5Hz，1H)，7.46(t，J＝7.5Hz，1H)，8.22(d，J＝7.5Hz，1H)，8.25(bs，1H)，8.65(s，2H)，10.57(bs，1H)。
the following compounds were obtained according to a similar method:
6-cyclopropyl-2- {3- [5- (1-methylpiperidin-4-yloxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one
2- {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzyl } -6-isopropyl-2H-pyridazin-3-one formate salt
6-cyclopropyl-2- {3- [5- (piperidin-4 yloxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one (prepared by Boc-protected compound)
6-cyclobutyl-2- {3- [5- (3-dimethylaminopropoxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one
6-cyclopropyl-2- {3- [5- (2-morpholin-4-ylethoxy) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one
Example 6
The preparation of 6-cyclopropyl-2- {3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("a 12") was carried out analogously according to the following scheme:
6.1A 500ml aqueous solution of 30g (215mmol) of potassium carbonate is added to a solution of 23.3g (82.1mmol) of methyl 3- [5- (dimethylaminomethyleneamino) pyrimidin-2-yl ] benzoate in 300ml dioxane and the mixture is stirred at 100 ℃ for 12 hours. The reaction mixture was cooled to room temperature and the pH was adjusted to 6-7 with 35ml of 37% aqueous hydrochloric acid. The solution was dried essentially in vacuo and lyophilized (crude 3- (5-aminopyrimidin-2-yl) benzoic acid; ESI 216). 400ml of methanol were added to the residue and 26ml of concentrated sulfuric acid were added dropwise to the suspension obtained in the ice bath. The reaction mixture was stirred at room temperature for 6 hours, then 500ml water was added and 32% aqueous sodium hydroxide was added in an ice bath until the pH was basic. The mixture was extracted with ethyl acetate. The organic phase was dried over sodium sulfate and evaporated: 3- (5-aminopyrimidin-2-yl) benzoic acid methyl ester as a brown solid; the ESI 230.
6.2 2 to a solution of 2.50g (10.9mmol) of methyl 3- (5-aminopyrimidin-2-yl) benzoate in 10ml of 1-methyl-2-pyrrolidinone 2.59g (1.7mmol) of potassium carbonate and 3.6g (1.7mmol) of bis (2-chloroethyl) methylammonium chloride are added, the suspension is stirred under argon at 120 ℃ for 18 h and at 140 ℃ for 6 h. The reaction mixture was cooled to room temperature, 150ml of water was added, and the mixture was filtered through celite. The filtrate was adjusted to pH 14 with 32% sodium hydroxide solution and extracted twice with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated: 3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] benzoic acid methyl ester as a brown oil; ESI 313.
6.3 at room temperature, 13.8ml of a 1M THF solution of diisobutylaluminum hydride are added dropwise to a solution of 860mg (2.75mmol) of methyl 3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] benzoate in 16ml of THF, which is kept under an argon atmosphere. To the reaction mixture was added 3ml of a saturated sodium sulfate solution and dichloromethane. The precipitate formed is filtered off with suction and washed with dichloromethane. The filtrate was dried over sodium sulfate and evaporated: {3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] phenyl } methanol as a yellow solid; ESI 285.
6.4 Add 402mg (equivalent to 1.2mmol) of polymer-bound triphenylphosphine and 277mgg (1.20mmol) of di-tert-butyl azodicarboxylate to a solution of 54.7mg (0.402mmol) of 6-cyclopropyl-2H-pyridazin-3-one and 114mg (0.402mmol) of {3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] phenyl } methanol in 3ml of THF and 1ml of DMF kept under an argon atmosphere and stir the mixture at room temperature for 3 hours. The reaction mixture was filtered through celite with suction. The filtrate was evaporated and purified by preparative HPLC: 6-cyclopropyl-2- {3- [5- (4-methylpiperazin-1-yl) pyrimidin-2-yl ] benzyl } -2H-pyridazin-3-one ("A12") bistrifluoroacetate as a colorless oil; ESI 403.
Example 7
Preparation of 6-isopropyl-2- (3- {5- [ 1- (2-morpholin-4-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one ("a 13") was carried out according to the following scheme:
7.1A mixture of 14.4g (157mmol) of glyoxylic acid monohydrate and 50ml (464mmol) of isopropyl methyl ketone is heated at 120 ℃ for 2 hours with stirring. The reaction mixture was cooled to 40 ℃ and 70ml of water and 12ml of 32% aqueous ammonia solution were added. The mixture was extracted three times with dichloromethane. 7.55ml (155mmol) of hydrazine hydroxide are added to the aqueous phase and the mixture is heated at reflux for 18 hours. The reaction mixture was cooled to room temperature and extracted with dichloromethane. The organic phase was dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography using petroleum ether/tert-butyl methyl ether as eluent to give 4-hydroxy-5, 5, 6-trimethyl-4, 5-dihydro-2H-pyridazin-3-one as colorless crystals (ESI 157) and 6-isopropyl-2H-pyridazin-3-one as colorless crystals (ESI 139).
The following compounds were also prepared using this method: 6-cyclobutyl-2H-pyridazin-3-one.
7.2 to a solution of 2.65g (10.0mol) [3- (5-bromopyrimidin-2-yl) phenyl ] methanol and 3.38g (11.0mmol)4- {2- [ 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazol-1-yl ] ethyl } morpholine in 50ml 1, 2-dimethoxyethane maintained under a nitrogen atmosphere, 4.25g (20.0mmol) of tripotassium phosphate trihydrate and 842mg (1.2mmol) of bis (triphenylphosphine) palladium chloride are added and the mixture is stirred at 80 ℃ for 18 hours. Dichloromethane and water were added to the reaction mixture, which was then filtered through celite, and the residue was washed with dichloromethane. The organic phase was separated, dried over sodium sulfate and evaporated. The residue was recrystallized from 2-propanol to give (3- {5- [ 1- (2-morpholin-4-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } phenyl) methanol as pale yellow crystals; ESI 366.
7.3A solution of 324mg (0.70mmol) of (3- {5- [ 1- (2-morpholin-4-ylethyl) -1H-pyrazol-4-yl ] -pyrimidin-2-yl } phenyl) methanol, 131mg (0.91mmol) of 6-isopropyl-2H-pyridazin-3-one and 278mg (1.05mmol) of triphenylphosphine in 1.4ml of THF is cooled in an ice bath under nitrogen and 217. mu.l (1.056mmol) of diisopropyl azodicarboxylate is added dropwise. After stirring the reaction mixture at room temperature for 2 hours, it was evaporated. The residue was purified by silica gel column chromatography using dichloromethane/methanol as eluent to give 6-isopropyl-2- (3- {5- [ 1- (2-morpholin-4-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one ("a 13") as pale yellow crystals; ESI 486;
1H-NMR (DMSO-d 6): δ [ ppm ] 1.20(d, J ═ 7Hz, 6H), 2.44(m, 4H), 2.77(t, J ═ 6.5Hz, 2H), 2.90 (heptad, J ═ 7Hz, 1H), 3.57(m, 4H), 4.30(t, J ═ 6.5Hz, 2H), 5.31(s, 2H), 6.97(d, J ═ 9.3Hz, 1H), 7.44(d, J ═ 7.5Hz, 1H), 7.50(t, J ═ 7.5Hz, 1H), 7.51(d, J ═ 9.3Hz, 1H), 8.11(s, 1H), 8.31(d, J ═ 7.5Hz, 1H), 8.36(bs, 1H), 8.44(s, 13H), 13.13H).
The following compounds were obtained according to a similar method:
2- (3- {5- [ 1- (2-Morpholin-4-ylethyl) -1H-pyrazol-4-yl ] pyrimidin-2-yl } benzyl) -2H-pyridazin-3-one ("A14") trifluoroacetate salt
Example 8
The preparation of 2- [3- (5-aminopyrimidin-2-yl) benzyl ] -6-cyclopropyl-2H-pyridazin-3-one ("a 18") was carried out analogously according to the following scheme. The individual reaction steps were carried out analogously to the corresponding reaction methods in examples 5 and 6.
Pharmacological data
TABLE 1Met kinase inhibition (enzyme assay and/or cellular assay)
| Compound numbering | IC50(enzyme) | IC50(cell) |
| ″A1″ | A | B |
| ″A2″ | A | A |
| ″A3″ | A | A |
| ″A4″ | A | B |
| ″A5″ | A | A |
| ″A6″ | A | B |
| ″A7″ | B | C |
| ″A8″ | A | A |
| ″A9″ | A | A |
| ″A10″ | A | B |
| ″A11″ | A | B |
| ″A12″ | A | A |
| ″A13″ | A | B |
| ″A14″ | A | B |
| ″A15″ | A | A |
| ″A18″ | B | B |
lC50：1nM-0.1μM＝A 0.1μM-10μM＝B＞10μM＝C
The following examples relate to pharmaceuticals:
example A: injection vial
A solution of 100g of the active ingredient of the formula (I) and 5g of disodium hydrogen phosphate in 3l of bidistilled water is adjusted to pH 6.5 using 2N hydrochloric acid, sterile-filtered, transferred into injection vials, freeze-dried under sterile conditions and sealed under sterile conditions. Each injection vial contained 5mg of active ingredient.
Example B: suppository
A mixture of 20g of active ingredient of formula (I) with 100g of soya lecithin and 1400g of cocoa butter was melted, poured into moulds and allowed to cool. Each suppository contains 20mg of active ingredient.
Example C: solutions of
Preparation of 1g of active ingredient of the formula (I), 9.38g of NaH2PO4·2H2O、28.48g Na2HPO4·12H2O and 0.1g benzalkonium chloride in 940ml double distilled water. The pH was adjusted to 6.8, the solution was made up to 1l and sterilized by irradiation. The solution may be used in the form of eye drops.
Example D: ointment formulation
500mg of active ingredient of formula (I) are mixed with 99.5g of vaseline under sterile conditions.
Example E: tablet formulation
A mixture of 1kg of active ingredient of formula (I), 4kg of lactose, 1.2kg of potato starch, 0.2kg of talc and 0.1kg of magnesium stearate is compressed in a conventional manner into tablets, so that each tablet contains 10mg of active ingredient.
Example F: dragee (dragee)
Tablets were compressed in a similar manner to example E and subsequently coated in a conventional manner with a coating of sucrose, potato starch, talc, tragacanth and dye.
Example G: capsule preparation
2kg of active ingredient of formula (I) are filled into hard gelatin capsules in a conventional manner so that each capsule contains 20mg of active ingredient.
Example H: ampoule agent
A solution of 1kg of the active ingredient of the formula (I) in 60l of bidistilled water is sterile-filtered, transferred into ampoules, freeze-dried under sterile conditions and sealed under sterile conditions. Each ampoule contains 10mg of active ingredient.
Claims (16)
1. Compounds of formula (I) and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios:
wherein:
R1represents a group of formula (I) wherein R represents H or A,
R2represents furyl, thienyl, pyrrolyl, imidazolyl, pyrazoleA base,Azolyl radical, isoOxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxazolyl, thiazolyl, triazolyl, tetrazolyl,oxadiazolyl or thiadiazolyl radical, each of which is substituted by Hal, A, [ C (R)3)2]nN(R3)2、[C(R3)2]nHet、O[C(R3)2]nN(R3)2And/or O [ C (R) ]3)2]nHet is singly substituted by a substituent group (Het),
R3represents H, methyl, ethyl or propyl,
R4、R4’represents a compound of formula (I) or (II),
het represents piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl,Azolyl radical, isoOxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, triazolyl, tetrazolyl, oxazolyl, thiazolyl, triazolyl, tetrazolyl,oxadiazolyl, thiadiazolyl, pyridazinyl or pyrazinyl, each of which is substituted by A or [ C (R)3)2]nHet1The process is a single substitution process,
Het1represents pyrrolidine, piperidine, piperazine or morpholine, each of which is unsubstituted or mono-or disubstituted by A and/or ═ O (carbonyl oxygen),
a represents unbranched or branched alkyl having 1 to 8C atoms, in which 1 to 7H atoms may be replaced by F and/or Cl, or represents cycloalkyl having 3 to 7C atoms,
hal represents F, Cl, Br or I,
n represents 0, 1, 2, 3 or 4.
2. A compound according to claim 1, selected from the following compounds, and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios:
3. a process for the preparation of a compound of formula (I) according to claim 1 or 2, and pharmaceutically acceptable salts and tautomers thereof, characterized in that:
so that the compound of the formula (II)
Wherein R is1As defined in claim 1, with a compound of formula (III):
wherein R is2、R3、R4And R4’As defined in claim 1, and
l represents Cl, Br, I or a free OH group,
and/or
Converting the base of formula (I) into one of its salts.
4. A medicament comprising at least one compound of the formula (I) according to claim 1 or 2 and/or pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, optionally together with auxiliary agents.
5. The medicament of claim 4, wherein the adjuvant is an excipient.
6. Use of a compound according to claim 1 or 2, its pharmaceutically acceptable salts, stereoisomers and mixtures thereof, including all ratios, for the preparation of a medicament for the treatment of diseases in which inhibition of tyrosine kinases by the compounds according to claim 1 or 2 has an effect.
7. Use of a compound according to claim 1 or 2, its pharmaceutically acceptable salts, stereoisomers and mixtures thereof, including all ratios, for the manufacture of a medicament for the treatment of diseases in which inhibition of Met kinase by a compound according to claim 1 or 2 has an effect.
8. The use according to claim 6 or 7, wherein the disease to be treated is a solid tumor.
9. The use of claim 8, wherein the solid tumor is derived from a tumor of squamous epithelium, bladder, stomach, kidney, head and neck, esophagus, cervix, thyroid, intestine, liver, brain, prostate, genitourinary tract, lymphatic system, stomach, larynx and/or lung.
10. The use of claim 8, wherein the solid tumor is derived from lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma, and breast cancer.
11. The use according to claim 6 or 7, wherein the disease to be treated is monocytic leukemia.
12. The use of claim 8, wherein the solid tumor is derived from lung adenocarcinoma, small cell lung carcinoma, pancreatic carcinoma, glioblastoma, colon carcinoma, and breast carcinoma.
13. The use of claim 6 or 7, wherein the disease to be treated is a tumor of the blood and immune system.
14. The use of claim 13, wherein the tumor is derived from acute myeloid leukemia, chronic myeloid leukemia, acute lymphatic leukemia and/or chronic lymphatic leukemia.
15. A medicament comprising at least one compound of the formula (I) and/or pharmaceutically acceptable salts and stereoisomers thereof, including mixtures thereof in all ratios, according to claim 1 or 2, and also comprising at least one further pharmaceutically active ingredient.
16. A kit consisting of the following individual packages:
(a) an effective amount of a compound of formula (I) according to claim 1 or 2 and/or its pharmaceutically acceptable salts and stereoisomers, including mixtures thereof in all ratios,
and
(b) an effective amount of other pharmaceutically active ingredients.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE102007061963.6 | 2007-12-21 | ||
| DE102007061963A DE102007061963A1 (en) | 2007-12-21 | 2007-12-21 | pyridazinone derivatives |
| PCT/EP2008/009970 WO2009083076A1 (en) | 2007-12-21 | 2008-11-25 | 2-benzylpyridazinone derivatives as met kinase inhibitors |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1151291A1 HK1151291A1 (en) | 2012-01-27 |
| HK1151291B true HK1151291B (en) | 2014-05-02 |
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