HK1186179B

HK1186179B - Pyrrolidinones as metap2 inhibitors

Info

Publication number: HK1186179B
Application number: HK13113621.5A
Authority: HK
Inventors: Timo Heinrich; Frank Zenke; Michel Calderini; Djordje Musil
Original assignee: Merck Patent Gmbh
Priority date: 2010-10-13
Filing date: 2011-09-14
Publication date: 2017-04-21

Description

Pyrrolidones as MetAP-2 inhibitors

The present invention relates to compounds of formula I and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios,

wherein

R¹Means (CH)₂)_nAr¹、(CH₂)_nHet、(CH₂)_nCyc、

Or a combination of A and B, or a combination of A,

R²means (CH)₂)_nAr²、CH[B(OH)₂]CH₂Het、

CH (C.ident.CH) phenyl, (CH)₂)_nHet、(CH₂)_nThe combination of Cyc and A is shown in the specification,

R³is OH, Hal, NH₂、CN、CF₃、CHF₂Or N₃，

R⁴H or alkyl having 1,2,3 or 4C atoms,

R²and R⁴Together also denotes alkylene having 2,3,4 or 5C atoms, one CH of which₂The groups may also be substituted by NH, NA, N-COA, N- (CH)₂)_nAr³、N-(CH₂)_nHet²、CH-A、CH-O-(CH₂)_nAr³、N-SO₂A or O is replaced by the N-substituted aryl,

and/or may be substituted by a,

Ar¹refers to phenyl, naphthyl or biphenyl, which is unsubstituted or mono-, di-, tri-, tetra-or pentasubstituted by: hal, A, OH, OA, NH₂、NHA、NA₂、NO₂、CN、COOH、COOA、CONH₂、CONHA、CONA₂、NHCOA、NHSO₂A、COA、CHO、Het¹、SO₂NH₂And/or SO₂A，

Ar²Refers to phenyl, naphthyl or biphenyl, which is unsubstituted or mono-, di-, tri-, tetra-or pentasubstituted by: hal, A, OH, OA, NH₂、NHA、NA₂、NO₂、CN、COOH、COOA、CONH₂、CONHA、CONA₂、NHCOA、NHSO₂A、COA、CHO、Het¹、SO₂NH₂And/or SO₂A，

Ar³Phenyl which is unsubstituted or mono-, di-or trisubstituted by Hal, OH, OA and/or A,

het denotes a mono-or bicyclic saturated, unsaturated or aromatic heterocycle having 1 to 4N-and/or O-and/or S-atoms, which is unsubstituted or mono-, di-or trisubstituted by: hal, A, OH, OA, NH₂、NHA、NA₂、NO₂、CN、COOH、COOA、(CH₂)_nCONH₂、(CH₂)_nCONHA、(CH₂)_nCONA₂、NHCOA、NHSO₂A、COA、CHO、Het¹、SO₂NH₂、SO₂A and/or = O,

Het¹means having a value of 1 toMonocyclic saturated, unsaturated or aromatic heterocycles of 4N-and/or O-and/or S-atoms, unsubstituted or substituted by Hal, A, OH, OA, NH₂、NHA、NA₂、NO₂CN, COOH and/or COOA mono-, di-or trisubstituted,

Het²refers to pyridyl, pyrimidyl, furyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl or thiadiazole,

a is a straight-chain or branched alkyl radical having 1 to 10C atoms, where 1 to 7H atoms may be replaced by F, Cl, Br, OH, CHO, COA, COOA, CN, CONA₂CONHA and/or CONH₂Instead of this, the user can,

and/or one or two non-adjacent CH-and/or CH₂The radicals may be O, N and/or NR⁴Instead of this, the user can,

or a group of cells of the group Cyc,

cyc means cycloalkyl having 3 to 7C atoms,

hal is F, Cl, Br or I,

n is 0,1, 2,3 or 4.

The present invention is based on the object of finding novel compounds having valuable properties, in particular those which can be used for the preparation of medicaments.

It has been found that the compounds of formula I and their salts have very valuable pharmacological properties while being well tolerated.

They exhibit a modulating, regulating and/or inhibiting effect in particular on metalloproteinases, preferably on methionine aminopeptidase (MetAP), in particular on the subtype MetAP-2.

They are useful as anticancer agents, as agents that positively influence fat metabolism, and as anti-inflammatory agents.

It has been found that the S enantiomer of the compounds according to the invention is significantly more active against MetAP-2 than the mirror image (R enantiomer).

Other hydroxy-substituted pyrrolidones are known from the following documents:

Zeitschrift für Naturforschung, B: ChemicalSciences (1994), 49(11), 1586-95;

Analytica ChimicaActa (1987), 202, 167-74;

Journal of ElectroanalyticalChemistry and Interfacial Electrochemistry (1988), 239(1-2), 161-73;

part B of Zeitschrift fur Naturfor. Chem Anorg. Chem (1978),33B (12), 1540-6;

J. Chem. Soc. (1965), (Oct.),5556-62;

J. Chem. Soc. (1965), (Oct.), 5551-6。

substituted hydrazides and N-alkoxyamides having MetAP-2 inhibitory activity and useful for inhibiting angiogenesis, particularly for treating diseases whose development is dependent on angiogenesis, such as cancer, are described in WO 01/79157.

WO 02/081415 describes MetAP-2 inhibitors useful in the treatment of cancer, hemangiomas, proliferative retinopathies, rheumatoid arthritis, atherosclerotic neovascularization, psoriasis, ocular neovascularization and obesity.

Compounds which are angiogenesis inhibitors and MetAP-2 inhibitors useful in the treatment of lymphoid leukemias and lymphomas are described in WO 2008/011114.

The anticancer effect of the compounds according to the invention lies in particular in their antiangiogenic effect. Angiogenesis inhibition has proven useful in more than 70 diseases, such as ovarian Cancer (f. Spinella et al j. cardiovasc. pharmacol.2004, page 44, 140), breast Cancer (a. Mora-bito et al crit. rev. oncol./hematol.2004, 49, 91), prostate Cancer (b. Nicholson et al Cancer metastas. rev.2001, 20, 297), diabetic blindness, psoriasis and macular degeneration (e. Ng et al can j. ophthalmol. 2005,23, 3706).

Proteases regulate many different cellular processes, in particular the regulation of peptides and proteins, in particular protein conversion, protein maturation and signal peptide processing, the breakdown of abnormal proteins and the inactivation/activation of regulatory proteins. In particular, amino-terminal modification of nascent polypeptides is the most common adjustment. Aminoproteases are metalloproteinases that cleave the unprotected N-terminal amino acid of a peptide or protein, either co-translationally (co-translational) or post-translationally (post-translational). .

Methionine aminopeptidase (MetAP) cleaves the terminal methionine of nascent peptides, particularly if the penultimate amino acid is small and uncharged (e.g., Gly, Ala, Ser, Thr, Val, Pro, or Cys).

In many disease processes, angiogenesis is the pathogenic center of the disease or has an exacerbating effect on the development of the disease. In the case of cancer, for example, angiogenesis can lead to enlargement of tumors and entry into other organs. Other diseases in which angiogenesis plays an important role are psoriasis, arthrosis, arteriosclerosis and ocular diseases, such as diabetic retinopathy, age-induced macular degeneration, rubeosis iridis or neovascular glaucoma, and inflammation. Thus, the compounds of formula I, compositions comprising these compounds and the methods underlying the present invention are useful in the treatment of these diseases.

Accordingly, the compounds of the present invention, or pharmaceutically acceptable salts thereof, are useful in the treatment of cancer, including solid cancers, such as cancers of the lung, pancreas, thyroid, bladder or colon, myelopathies (e.g., myelogenous leukemia) or adenomas (e.g., villous colon adenocarcinoma).

Tumors also include monocytic leukemia, brain cancer, genitourinary cancer, lymphatic cancer, stomach cancer, laryngeal cancer, and lung cancer including lung adenocarcinoma and small cell lung cancer, pancreas and/or breast cancer.

The present invention therefore relates to compounds according to the invention as medicaments and/or medicament active ingredients in the treatment and/or prophylaxis of said diseases and to the use of compounds according to the invention for the production of medicaments for the treatment and/or prophylaxis of said diseases, as well as to methods for the treatment of said diseases, comprising the administration of one or more compounds according to the invention to a patient in need of such administration.

It can be demonstrated that the compounds according to the invention have an anticancer effect. The compounds of the invention are administered to a patient suffering from a disease, for example, for inhibiting tumor growth, for reducing inflammation associated with lymphoproliferative diseases, for inhibiting transplant rejection or nerve damage resulting from tissue repair, and the like. The compounds are useful for prophylactic or therapeutic purposes. The term "treatment" as used herein is intended to mean both prevention of disease and treatment of existing symptoms. Prevention of proliferation/survival, e.g. for preventing tumor growth, is achieved by administering a compound according to the invention prior to the development of a dominant disease. Alternatively, the compounds are useful for treating persistent disorders by stabilizing or ameliorating the clinical symptoms of a patient.

The host or patient may belong to any mammalian species, for example a primate species, particularly humans; rodents, including mice, rats, and hamsters; rabbits; horses, cattle, dogs, cats, etc. Animal models are of interest for experimental studies, where they provide a model for the treatment of human diseases.

The susceptibility of a particular cell to treatment with a compound according to the invention can be determined by in vitro assays. Cell cultures are generally incubated with the compounds according to the invention at various concentrations for a sufficient period of time, often between about 1 hour and 1 week, for the active agent to induce cell death or inhibit cell proliferation, cell viability or migration. Cells cultured from biopsy samples can be used for in vitro assays. The amount of cells remaining after treatment was then determined.

The dosage will vary with the particular compound employed, the particular disease, the condition of the patient, and the like. The therapeutic dose is generally sufficient to substantially reduce the undesirable cell population in the target tissue while maintaining the viability of the patient. Treatment is generally continued until there is a substantial reduction in cell load, e.g., at least about a 50% reduction in cell load, and treatment is continued until substantially no more undesired cells are detected in the body.

It has been found that the compounds according to the invention cause specific inhibition of MetAP-2. The compounds according to the invention preferably exhibit advantageous biological activities which are detectable in assays such as those described herein. In such assays, the compounds according to the invention exhibit and cause an inhibitory effect, often consisting of an IC in the appropriate range, preferably in the micromolar range and more preferably in the nanomolar range₅₀The values are represented.

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

The compounds according to the invention can also be used for the treatment of obesity (adiposity). Henri R. Lijnen et al, Obesity, Vol.18, No.12, 2241-2246 (2010) describe the use of a Met-AP2 inhibitor, fumagillin, in the reduction of adipose tissue.

The use of Met-AP2 inhibitors (fumagillin type compounds) for the treatment of obesity is also described in WO 2011/085201 a 1.

The compounds according to the invention are also useful in the treatment of malaria. The use of a Met-AP2 inhibitor, fumagillin, in the treatment of malaria is described by Chem et al in Chemistry & Biology, Vol.16, 193-202 (2009).

The compounds according to the invention can also be used for the treatment of benign prostatic hyperplasia.

The use of Met-AP2 inhibitors (fumagillin type compounds) for the treatment of benign prostatic hyperplasia is described in WO 2011/085198 a 1.

The compounds of formula I are also intended to mean the hydrates and solvates of these compounds, as well as the pharmaceutically acceptable derivatives.

The invention also relates to optically active forms (stereoisomers), salts, enantiomers, racemates, diastereomers and hydrates and solvates of these compounds. Solvates of the compounds are understood to be additions of inert solvents to the compounds which are formed as a result of their mutual attractive forces. Solvates are, for example, mono-or dihydrate or alcoholates.

Pharmaceutically acceptable derivatives mean, for example, salts of the compounds according to the invention and also the so-called prodrug compounds.

Prodrug derivatives mean compounds of the formula I which have been modified with, for example, alkyl or acyl groups, sugars or oligopeptides and which are rapidly cleaved in an organism to give the effective compounds according to the invention.

These also include, as for example, in int. j. Pharm.115Biodegradable polymer derivatives of the compounds according to the invention described in 61-67 (1995).

The term "effective amount" means that amount of an agent or pharmaceutically active ingredient that causes a biological or medical response in a tissue, system, animal or human that is being sought or desired, for example, by a researcher or physician.

Furthermore, the term "therapeutically effective amount" refers to an amount that has the following consequences compared to a corresponding subject not receiving such an amount: improved treatment, healing, prevention or elimination of a disease, condition, disease state, disorder or side effect or lessening the development of a disease, disorder or condition.

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

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

These are particularly preferably mixtures of stereoisomeric compounds.

The invention relates to compounds of formula (I) and salts thereof, and to processes for the preparation of compounds of formula (I) and pharmaceutically acceptable salts, tautomers and stereoisomers thereof, characterized in that

a) Reacting a compound of formula II

Wherein R is¹And R³Having the meaning indicated in claim 1,

and L denotes Cl, Br, I or a free or reactively functionally modified OH group,

with compounds of the formula III

Wherein R is²And R⁴Having the meaning indicated in claim 1,

or

b) To prepare a compound in which R is³Refers to a compound of formula I of OH,

oxidizing a compound of formula IV

Wherein R is¹、R²And R⁴Having the meaning indicated in claim 1,

or

c) By replacing OH groups by halogen atoms or by N₃Instead of halogen atoms, with radicals R³Conversion to another radical R³，

And/or converting the base or acid of formula I into one of its salts.

In this context, except forUnless explicitly stated otherwise, the residue R¹、R²、R³Have the meaning indicated in formula I.

A is alkyl, straight-chain (linear) or branched and has 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-, 1, 2-or 2, 2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3-or 4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2, 3-or 3, 3-dimethylbutyl, 1-or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1, 2-or 1,2, 2-trimethylpropyl, preferably trifluoromethyl.

A preferably means a straight-chain or branched alkyl radical having 1 to 6C atoms, where 1 to 7H atoms may be replaced by F, Cl, Br and/or OH,

or Cyc.

Furthermore, A preferably means a straight-chain or branched alkyl radical having 1 to 10C atoms, where 1 to 7H atoms may be replaced by F, Cl, Br, OH, CHO, COA, COOA, CN, CONA₂CONHA and/or CONH₂Instead of this, the user can,

or Cyc.

A very particularly preferably means 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.

Cycloalkyl is preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.

R¹Preferably phenyl, benzyl, naphthyl or biphenyl, which is unsubstituted or substituted by Hal, CN, NHCOA, NHSO₂A、SO₂A and/or CONH₂Mono-, di-, tri-, tetra-or penta-substituted;

a or (CH)₂)_nHet。

R²Preferably, [ C (R)⁴)₂]_nAr²、(CH₂)_nCyc、

CH (C.ident.CH) phenyl, A or (CH)₂)_nHet。

R³Preferably OH and N₃、NH₂Or F.

R⁴Preferably H, methyl, ethyl or propyl.

Ar¹Is understood to mean, for example, phenyl, o-, m-or p-tolyl, o-, m-or p-ethyl-phenyl, o-, m-or p-propylphenyl, o-, m-or p-isopropylphenyl, o-, m-or p-tert-butylphenyl, o-, m-or p-trifluoromethyl-phenyl, o-, m-or p-fluorophenyl, o-, m-or p-bromophenyl, o-, m-or p-chlorophenyl, o-, m-or p-hydroxyphenyl, o-, m-or p-methoxyphenyl, o-, m-or p-methylsulfonylphenyl, o-, m-or p-nitrophenyl, o-, m-or p-aminophenyl, o-, m-or p-methylaminophenyl, M-or p-dimethyl-aminophenyl, o-, m-or p-aminosulfonylphenyl, o-, m-or p-aminocarbonylphenyl, o-, m-or p-carboxyphenyl, o-, m-or p-methoxycarbonylphenyl, o-, m-or p-ethoxycarbonylphenyl, o-, m-or p-acetylphenyl, o-, m-or p-formylphenyl, o-, m-or p-cyanophenyl,

further preferred are 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-, 2,5-, 2,6-, 3, 4-or 3, 5-dibromophenyl, 2,3,4-, 2,3,5-, 2,3,6-, 2,4, 6-or 3,4, 5-trichlorophenyl, p-iodophenyl, 4-fluoro-3-chlorophenyl, 2-fluoro-4-bromophenyl, 2, 5-difluoro-4-bromophenyl or 2, 5-dimethyl-4-chlorophenyl; furthermore naphthyl or biphenyl.

Ar¹Also preferred is phenyl, naphthyl or biphenyl, which is unsubstituted or mono-, di-, tri-, tetra-or pentasubstituted by Hal.

Ar²Is understood to mean, for example, phenyl, o-, m-or p-tolyl, o-, m-or p-ethyl-phenyl, o-, m-or p-propylphenyl, o-, m-or p-isopropylphenyl, o-, m-or p-tert-butylphenyl, o-, m-or p-trifluoromethyl-phenyl, o-, m-or p-fluorophenyl, o-, m-or p-bromophenyl, o-, m-or p-chlorophenyl, o-, m-or p-hydroxyphenyl, o-, m-or p-methoxyphenyl, o-, m-or p-methylsulfonylphenyl, o-, m-or p-nitrophenyl, o-, m-or p-aminophenyl, o-, m-or p-methylaminophenyl, M-or p-dimethyl-aminophenyl, o-, m-or p-aminosulfonylphenyl, o-, m-or p-aminocarbonylphenyl, o-, m-or p-carboxyphenyl, o-, m-or p-methoxycarbonylphenyl, o-, m-or p-ethoxycarbonylphenyl, o-, m-or p-acetylphenyl, o-, m-or p-formylphenyl, o-, m-or p-cyanophenyl,

Ar²Preference is also given to phenyl which is unsubstituted or mono-, di-, tri-, tetra-or pentasubstituted by A, Hal, CN, OH and/or OA.

Het, irrespective of further substituents, denotes, 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, preferably furthermore 1,2, 3-triazol-1-, -4-or-5-yl, optionally substituted phenyl, 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, 1-, (meth) acrylic acid or methacrylic acid, 2-, 4-or 5-benzimidazolyl, 1-, 2-, 3-, 4-, 5-, 6-or 7-indazolyl, 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-, 5-, 6-or 7-benz-zoxazolyl, 2-, 4-, 5-, 6-or 7-benz-zoxazolyl, 3-, 4-, 5-, 6-, 7-or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-or 8-isoquinolyl, 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, further preferably 1, 3-benzodioxol-5-yl, 1, 4-benzodioxan-6-yl, 2,1, 3-benzothiadiazol-4-or-5-yl or 2,1, 3-benzoxadiazol-5-yl.

The heterocyclic residue may also be partially or fully hydrogenated.

Unsubstituted 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, 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-morpholinyl, 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-ethylenedioxyphenyl, isoindolyl, 3,4- (difluoro-methylenedioxy) -phenyl, 2, 3-dihydro-benzofuran-5-or 6-yl, 2,3- (2-oxomethylenedioxy) -phenyl or 3, 4-dihydro-2H-1, 5-benzodioxepin-6-or-7-yl, further preferably 2, 3-dihydro-benzo-furanyl or 2, 3-dihydro-2-oxofuranyl.

Het furthermore preferably denotes pyridazinyl, pyrazolyl, benzimidazolyl, pyridyl, dibenzofuranyl, carbazolyl, indolyl, dihydro-indolyl, benzofuranyl, dihydro-benzofuranyl, 2, 3-dihydro-benzo [1,4] benzo]Dioxinyl, chromanyl, piperazinyl, morpholinyl, tetrahydropyranyl, quinolinyl, isoquinolinyl, isoindolyl, dihydro-quinolinyl, dihydroisoquinolinyl, tetrahydro-quinolinyl, tetrahydroisoquinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pyrimidinyl, indazolyl, furanyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazole, benzothiazolyl, imidazo [1,2-a]Pyridyl, 1, 3-benzodioxolyl, benzoxazolyl, piperidin-1-yl, pyrrolidin-1-yl, [1, 2]]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or substituted by A, OA, COOA, COA, CHO, (CH)₂)_nCONH₂、SO₂A、NHSO₂A. = O and/or Het¹Mono-, di-or tri-substituted.

Het is particularly preferably dihydro-Indolyl, benzofuranyl, dihydro-benzofuranyl, 2, 3-dihydro-benzo [1,4]]Dioxinyl, chromanyl, oxazolyl, oxadiazolyl, tetrazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, thienyl, furanyl, tetrahydropyranyl, pyrazolyl, pyridyl, benzothiazolyl, 2, 3-dihydro-benzo [1,4] benzo]Dioxinyl, quinolyl, isoquinolyl or pyrrolidinyl, unsubstituted or substituted by A, COOA, COA, CHO, (CH)₂)_nCONH₂、(CH₂)_nCONHA、(CH₂)_nCONA₂、SO₂A、NHSO₂A. = O and/or pyrrolidinyl mono-or disubstituted.

Not taking into account further substituents, Het¹Means, 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, preferably also 1,2, 3-triazol-1-, -4-or 5-yl, 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 or pyrazinyl.

The heterocyclic group may also be partially or fully hydrogenated.

Unsubstituted Het¹Thus, 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, 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-morpholinyl, tetrahydro-2-, -3-or-4-pyranyl, 1, 4-dioxacyclohexyl, 1, 3-dioxan-2-, -4-or-5-yl, hexahydro-1-, -3-or-4-pyridazinyl, hexahydro-1-, -2-, -4-or-5-pyrimidinyl or 1-, 2-or 3-piperazinyl.

Het¹Furthermore preferred is pyridazinyl, pyrazolyl, pyridyl, piperazinyl, morpholinyl, pyrimidinyl, furanyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazole, piperidin-1-yl, pyrrolidin-1-yl, tetrahydro-pyranyl, [1,2]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or mono-, di-or trisubstituted by A and/or OA.

Het¹Particularly preferred is unsubstituted or monosubstituted by A de imidazolyl, thiazolyl or pyrrolidinyl.

Hal preferably means F, Cl or Br and I, particularly preferably F or Cl.

It is useful for the entire invention that all of the multiple occurrences of the residue may be the same or different, i.e., independent of each other.

The compounds of formula I may have one or more chiral centers and may therefore exist in various stereoisomeric forms. Formula I includes all of these forms.

The invention therefore relates in particular to those compounds of the formula I in which at least one of the residues has one of the preferred meanings indicated above. Some preferred groups of compounds can be represented by the following sub-formulae Ia to If, which correspond to formula I and in which the residues not specified in more detail have the meanings given in formula I, but in which

In Ia, R¹Refers to phenyl, benzyl, naphthyl or biphenyl, unsubstituted or substituted by Hal, CN, NHCOA, NHSO₂A、SO₂A and/or CONH₂Mono-, di-, tri-, tetra-or penta-substituted;

a or (CH)₂)_nHet；

In Ib, R²Means [ C (R)⁴)₂]_nAr²、(CH₂)_nCyc、CH[B(OH)₂]CH₂Het、

CH (C.ident.CH) phenyl, A or (CH)₂)_nHet；

In Ic, R³Is OH, N₃、NH₂Or F;

in Id, Het refers to pyridazinyl, pyrazolyl, benzimidazolyl, pyridyl, dibenzo-furyl, carbazolyl, indolyl, dihydro-indolyl, benzofuryl, dihydro-benzofuryl, 2, 3-dihydro-benzo [1,4] benzofuryl]Dioxinyl, chromanyl, piperazinyl, morpholinyl, tetrahydropyranyl, quinolinyl, isoquinolinyl, isoindolyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pyrimidinyl, indazolyl, furyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazole, benzothiazolyl, imidazo [1,2-a]Pyridyl, 1, 3-benzodioxolyl, benzoxazolyl, piperidin-1-yl, pyrrolidin-1-yl, [1, 2]]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or substituted by A, OA, COOA, COA, CHO, (CH)₂)_nCONH₂、(CH₂)_nCONHA、(CH₂)_nCONA₂、SO₂A、NHSO₂A. = O and/or Het¹Mono-, di-or tri-substituted;

in Ie, Het¹Is pyridazinyl, pyrazolyl, pyridyl, piperazinyl, morpholinyl, pyrimidinyl, furanyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, isoxazolylOxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazolyl, piperidin-1-yl, pyrrolidin-1-yl, tetrahydropyranyl, [1, 2]]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or mono-, di-or trisubstituted by A and/or OA;

in If, R¹Refers to phenyl, benzyl, naphthyl or biphenyl, unsubstituted or substituted by Hal, CN, NHCOA, NHSO₂A、SO₂A and/or CONH₂Mono-, di-, tri-, tetra-or penta-substituted;

a or (CH)₂)_nHet，

R²Means [ C (R)⁴)₂]_nAr²、CH[B(OH)₂]CH₂Het、

(CH₂)_nCyc、

CH (C.ident.CH) phenyl, A or (CH)₂)_nHet，

R³Is OH, N₃、NH₂Or F, the number of the first and second groups,

R⁴h or alkyl having 1,2,3 or 4C atoms,

and/or may be substituted by a,

het refers to pyridazinyl, pyrazolyl, benzimidazolyl, pyridyl, dibenzo-furyl, carbazolyl, indolyl, dihydro-indolyl and benzofuranPyran-yl, dihydro-benzofuran-yl, 2, 3-dihydro-benzo [1,4]]Dioxinyl, chromanyl, piperazinyl, morpholinyl, tetrahydropyranyl, quinolinyl, isoquinolinyl, isoindolyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pyrimidinyl, indazolyl, furyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazole, benzothiazolyl, imidazo [1,2-a]Pyridyl, 1, 3-benzodioxolyl, benzoxazolyl, piperidin-1-yl, pyrrolidin-1-yl, [1, 2]]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or substituted by A, OA, COOA, COA, CHO, (CH)₂)_nCONH₂、(CH₂)_nCONHA、(CH₂)_nCONA₂、SO₂A、NHSO₂A. = O and/or Het¹Mono-, di-or tri-substituted,

Het¹is pyridazinyl, pyrazolyl, pyridyl, piperazinyl, morpholinyl, pyrimidinyl, furanyl, thienyl, imidazolyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazole, piperidin-1-yl, pyrrolidin-1-yl, tetrahydropyranyl, [1, 2] methyl]Oxaazacyclohex-2-yl, [1,2, 5]]Oxadiazacyclohexane-2-yl, [1,3 ]]Oxaazacyclohex-3-yl or hexahydropyrimidyl, unsubstituted or mono-, di-or trisubstituted by A and/or OA,

and/or one or two non-adjacent CH and/or CH₂The radicals may be O, N and/or NR⁴Instead of this, the user can,

or a group of cells of the group Cyc,

Ar²refers to phenyl which is unsubstituted or mono-, di-, tri-, tetra-or pentasubstituted by A, Hal, CN, OH and/or OA,

cyc means cycloalkyl having 3 to 7C atoms,

hal is F, Cl, Br or I,

n is 0,1, 2,3 or 4;

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

Furthermore, the compounds of the formula I and the starting materials for their preparation are prepared by processes known per se, more precisely under reaction conditions which are known and suitable for the reaction, as described in the literature (for example in standard works, such as Houben-Weyl, Methoden der organischen Chemie, Georg-Thieme-Verlag, Stuttgart). Known per se solutions which are not mentioned in more detail in this context can also be used here.

The compounds of the formula I can preferably be obtained by reaction of compounds of the formula II with compounds of the formula III.

The compounds of formula II and formula III are generally known. However, if they are novel, they can be prepared by methods known per se.

In the compounds of the formula II, L preferably means Cl, Br, I or a free or reactively modified OH group, for example an activated ester, an imidazolyl (Imidazolid) or an alkylsulfonyloxy group having 1 to 6C atoms, preferably a methylsulfonyloxy or trifluoromethylsulfonyloxy group, or an arylsulfonyloxy group having 6 to 10C atoms, preferably a phenyl-or p-tolylsulfonyloxy group.

The reaction is preferably carried out in the presence of a dehydrating agent, optionally in the presence of N-hydroxy-benzotriazole, for example carbodiimides such as N, N ' -dicyclohexylcarbodiimide ("DCCI"), 1' -carbonyldiimidazole or N-3-dimethylaminopropyl-N ' -ethyl-carbodiimide ("DAPECI"), and propane phosphoric anhydride T3P (see Angew. chem. 92, 129 (1980)), diphenylphosphoryl azide or 2-ethoxy-N-ethoxycarbonyl-1, 2-dihydroquinoline;

T3P =。

the reaction is carried out in an inert solvent and 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 alkali metal-or alkaline earth metal-hydroxides, carbonates or bicarbonates, or else salts of weak acids of alkali metals or alkaline earth metals, preferably of potassium, sodium, calcium or cesium.

Depending on the conditions used, the reaction time is between a few minutes and 14 days and the reaction temperature is between about-15 ℃ and 150 ℃, generally between 40 ℃ and 130 ℃ and particularly preferably between 60 ℃ and 110 ℃.

Suitable as inert solvents are, for example, 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 ether or ethylene glycol monoethyl ether (ethylene glycol monomethyl ether or ethylene glycol 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.

Particularly preferred are glycol ethers, such as ethylene glycol monomethyl ether, THF, dichloromethane and/or DMF.

The compounds of formula I are also preferably obtained by oxidation of compounds of formula IV.

The oxidation is preferably carried out using tert-butyl hydroperoxide.

Water is preferred as solvent, wherein preferably an alkali metal-or alkaline earth metal-hydroxide, carbonate or bicarbonate, or an additional weak acid salt of an alkali metal or alkaline earth metal, preferably potassium, sodium, calcium or cesium, is added.

Pharmaceutically acceptable salts and other forms

The compounds according to the invention can be used in their final non-salt form. In another aspect, the invention also includes the use of these compounds in the form of their pharmaceutically acceptable salts, which can be derived from various organic and inorganic acids and bases according to routes known in the art. The pharmaceutically acceptable salt forms of the compounds of formula I are prepared mainly by conventional methods. If a compound of formula I contains a carboxylic acid group, one of its suitable salts may be formed by reacting the compound with a suitable base to produce 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; and various organic bases such as piperidine, diethanolamine and N-methylglutamine. Also included are aluminum salts of the compounds of formula I. In the case of certain compounds of formula I, acid addition salts may be formed by treating these 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, sulfates, nitrates or phosphates and the like and alkyl-and monoaryl sulfonates, such as ethanesulfonate, toluenesulfonate and benzenesulfonate, and other organic acids and their corresponding salts, such as acetate, trifluoroacetate, tartrate, maleate, succinate, citrate, benzoate, salicylate, ascorbate and the like. Accordingly, pharmaceutically acceptable acid addition salts of compounds of formula I include the following: acetate, adipate, alginate, arginate, aspartate, benzoate, benzenesulfonate (benzenesulfonate), bisulfate, bisulfite, bromide, butyrate, camphorate, camphorsulfonate, caprylate, chloride, chlorobenzoate, citrate, cyclo-pentanepropionate, digluconate, dihydrogenphosphate, dinitro-benzoate, dodecylsulfate, ethanesulfonate, fumarate, mucate (formed from mucic acid), galacturonate, glucoheptonate, gluconate, glutamate, glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate, lactobionate, malate, methosulfate, acetate, dihydrogenphosphate, dihydrogensulfate, camphorate, isovalerate, isobutyrate, lactate, lactobionate, malate, and mixtures thereof, Maleate, malonate, mandelate, metaphosphate, methanesulphonate, methyl-benzoate, monohydrogen phosphate, 2-naphthalenesulphonate, nicotinate, nitrate, oxalate, oleate, pamoate (Palmoat), pectinate, persulfate, phenylacetate, 3-phenylpropionate, phosphate, phosphonate, phthalate, but this is not limitative.

Furthermore, the basic salts of the compounds according to the invention include the aluminium-, ammonium-, calcium-, copper-, iron (III) -, iron (II) -, lithium-, magnesium-, manganese (III) -, manganese (II) -, potassium-, sodium and zinc salts, without this being intended to represent a limitation. Among the above salts, ammonium is preferred; alkali metal salts of sodium and potassium, and alkaline earth metal salts of calcium and magnesium. Salts of compounds of formula I derived from pharmaceutically acceptable organic toxophilic bases include primary, secondary and tertiary amines, substituted amines, also naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, chloroprocaine, choline, N' -dibenzyl-ethylenediamine (benzathine), dicyclohexyl-amine, diethanol-amine, diethyl-amine, 2-diethyl-amino-ethanol, 2-dimethyl-amino-ethanol, ethanol-amine, ethylenediamine, N-ethylmorpholine, N-ethyl-piperidine, reduced glucosamine, histidine, hydrabamine (Hydranamin), isopropylamine, lidocaine, lysine, meglumine, N-methyl-D-morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theine, triethanolamine, piperonyl alcohol, procaine, and also salts, Triethylamine, trimethylamine, tripropylamine and tri-hydroxy-methyl-amine (tromethamine) salts, but this is not intended to represent a limitation.

The compounds of the present invention containing basic nitrogen-containing groups may be used as (C)₁-C₄) Alkyl halides, such as methyl-, ethyl-, isopropyl-, and tert-butyl chloride, -bromo-, and-iodo; two (C)₁-C₄) Alkyl sulfates such as dimethyl-, diethyl-, and diamyl sulfates; (C)₁₀-C₁₈) Alkyl halides, such as decyl-, dodecyl-, lauryl-, tetradecyl-and octadecyl-chloride, -bromo and-iodo; and aryl- (C)₁-C₄) Alkyl halides, such as benzyl chloride and phenethyl bromide, are quaternized. Such salts may be used to prepare water-soluble and oil-soluble compounds according to the invention.

Preferred such pharmaceutically acceptable salts include 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, although this is not intended to represent a limitation.

Basic acid addition salts of the compounds of formula I are 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 certain respects, in certain physical properties, such as solubility in polar solvents; within the scope of the invention, however, the salts otherwise correspond to their respective free base forms.

As mentioned, pharmaceutically acceptable base addition salts of 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, di-ethanol-amine, ethylenediamine, N-methyl-D-glucamine and procaine.

The base addition salts of the acidic compounds according to the invention are prepared by contacting the free acid form with a sufficient amount of the desired base to form a salt in a conventional manner. The free acid may be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner. The free acid form differs from its corresponding salt form in certain respects, in certain physical properties, such as solubility in polar solvents; within the scope of the invention, however, the salts otherwise correspond to their respective free acid forms.

If a compound according to the invention contains more than one group capable of forming such a pharmaceutically acceptable salt, the invention also includes complex salts. Typical complex salt forms include, for example, bitartrate, diacetate, hydrogen fumarate, meglumine, diphosphate, disodium, and trihydrochloride, but this is not intended to represent a limitation.

As can be seen from the above, the term "pharmaceutically acceptable salt" means herein an active ingredient comprising a compound of formula I in the form of one of its salts, in particular if such 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 before. The pharmaceutically acceptable salt forms of the active ingredient may also confer for the first time the desired pharmacokinetic properties not previously present for this active ingredient and having a positive impact on the pharmacodynamics of this active ingredient even in its in vivo therapeutic efficacy.

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 preparations may be administered in dosage units containing a predetermined amount of the active ingredient per dosage unit. Depending on the disease state to be treated, the route of administration and the age, weight and condition of the patient, such units may contain, for example, from 0.5 mg to 1 g, preferably from 1 mg to 700 mg, particularly preferably from 5 mg to 100 mg, of a compound according to the invention, or the pharmaceutical preparations may be administered in dosage units which contain a predetermined amount of active ingredient per dosage unit. Preferred dosage unit formulations are those containing a daily dose or sub-dose, or corresponding fraction thereof, of the active ingredient as indicated above. In addition, such pharmaceutical preparations may be prepared using methods well known in the pharmaceutical art.

The pharmaceutical formulations may be adapted for administration via any suitable route, 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) means. Such formulations may be prepared, for example, by combining the active ingredient with one or more excipients or one or more adjuvants using all methods known in the pharmaceutical art.

Pharmaceutical formulations adapted for oral administration may be presented as discrete units, for example capsules or tablets; 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, in the case of oral administration in the form of tablets or capsules, the active ingredient may be combined with oral, non-toxic and pharmaceutically acceptable inert excipients, for example ethanol, glycerol, water and the like. Powders are prepared by grinding the compound to a suitable fine particle size and mixing it with a pharmaceutical excipient, e.g. an edible carbohydrate, such as starch or mannitol, which is ground in a similar manner. Flavoring, preservative, dispersing and coloring agents may also be present.

Capsules were made by preparing a powder mixture as described above and filling shaped gelatin shells with it. Glidants and lubricants, for example highly disperse silicon dioxide in solid form, talc, magnesium stearate, calcium stearate or polyethylene glycol, can be added to the powder mixture before the filling operation. Disintegrating or solubilizing agents, such as agar-agar, calcium carbonate or sodium carbonate, may also be added to improve the effectiveness of the medicament after the capsule is taken.

In addition, if desired or necessary, suitable binders, lubricants and disintegrants and also dyes can be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, sweeteners made from corn, natural and synthetic rubbers such as gum arabic, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or dry-compressing the mixture, adding a lubricant and a disintegrant and compressing them all into tablets. The powder mixture is prepared by mixing the compound comminuted in a suitable manner with a diluent or binder as described above and optionally with a binder, for example carboxymethylcellulose, alginates, gelatin or polyvinylpyrrolidone, a dissolution retardant, for example paraffin, an absorption promoter, for example a quaternary ammonium salt, and/or an absorbent, for example bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder, for example syrup, starch paste, acacia mucilage (Aacadia-Schleim) or a solution of cellulose-or polymer material and pressing it through a sieve. Instead of granulation, the powder mixture may be passed through a tablet press to produce a non-uniformly shaped mass which is broken up into granules. The particles may be greased by the addition of stearic acid, stearate, talc or mineral oil to prevent sticking to the cast sheet mould. The fat-coated mixture is then compressed into tablets. The compounds according to the invention can also be combined with free-flowing inert excipients and then compressed directly into tablets without a granulation or dry compression step. There may be a transparent or opaque protective layer consisting of a shellac sealing layer, a sugar or polymer material layer and a waxy gloss layer. 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 are prepared using a non-toxic alcoholic vehicle. Suspensions may be formulated by dispersing the compound in a non-toxic vehicle. Solubilizers and emulsifiers, such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers, preservatives, flavoring additives, such as peppermint oil, or natural or saccharin sweeteners or other artificial sweeteners, and the like, may also be added.

Dosage unit formulations for oral administration may optionally be encapsulated in microcapsules. The formulations may also be prepared in a prolonged or delayed release manner, for example by coating or embedding the particulate material in a polymer, wax or the like.

The compounds of the present invention and salts, solvates and physiologically functional derivatives thereof may also be administered in the form of liposome 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 to which the compound molecule is coupled. The compounds may also be coupled to soluble polymers as carriers for targeting agents. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenol, or polyethyleneoxide polylysine, substituted with palmitoyl residues. In addition, the compounds may be coupled to a class of biodegradable polymers suitable for achieving controlled release of the agent, such as crosslinked or amphiphilic block copolymers of polylactic acid, poly-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates and hydrogels.

Pharmaceutical formulations adapted for transdermal administration may be administered as a separate plaster in intimate contact with the epidermis of the recipient for an extended period of time. Thus, for example, iontophoresis may be used to deliver the active ingredient from a plaster, as described in general terms in Pharmaceutical Research,3(6), 318 (1986).

Pharmaceutical compounds suitable for topical administration may 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, such as the mouth and skin, the formulation is preferably applied in the form of a topical ointment or cream. In the case of formulation into an ointment, the active ingredient may be used with a paraffinic or water-miscible cream base. Alternatively, the active ingredient may be formulated in the form of a cream with an oil-in-water cream base or a water-in-oil base.

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

Pharmaceutical formulations suitable for topical application to the oral cavity include lozenges, pastilles and mouthwashes.

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

Pharmaceutical formulations suitable for nasal administration in which the carrier material is a solid include a coarse powder having a particle size of, for example, 20 to 500 microns which is administered by nasal inhalation, i.e. rapid inhalation through the nasal passage from a powder-containing container adjacent the nose. Formulations suitable for administration as nasal sprays or nasal drops in liquid form as carrier materials include solutions of the active ingredient in water or oil.

Pharmaceutical formulations adapted for administration by inhalation comprise a fine powder or mist which may be generated by means of various types of pressurised aerosol-containing dispensers, nebulisers or insufflators.

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

Pharmaceutical formulations adapted 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 may be included containing a suspending medium and a thickening agent. The formulations may be administered in unit-dose or multi-dose containers, for example sealed ampoules and vials, and stored in a freeze-dried (lyophilized) condition, so that only the addition of the sterile carrier liquid, for example water for injections, is required 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 ingredients specifically mentioned above, the formulation may also comprise other agents which are common in the art according to the 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, such as colon or breast cancer, will generally be in the range of from 0.1 to 100 mg/kg body weight of the recipient (mammal) per day, and particularly will generally be in the range of from 1 to 10 mg/kg body weight per day. Thus, an actual amount per day of an adult mammal weighing 70 kg is typically between 70 and 700 mg, wherein such amount may be administered as a single dose per day or typically in a series of divided doses per day (e.g. 2,3,4,5 or 6) such that the total daily dose is the same. The effective amount of the salt or solvate or physiologically functional derivative thereof can be determined as a ratio of the effective amount of the compound itself according to the present invention. Similar dosages are considered to be applicable for the treatment of the other disease conditions mentioned above.

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

The invention also relates to a kit of parts 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 suitable containers such as boxes or cartons, individual bottles, bags or ampoules. The kit may for example comprise separate ampoules, each containing an effective amount of a compound of formula I and/or pharmaceutically acceptable salts and stereoisomers thereof, including mixtures thereof in all ratios, and an effective amount of the other pharmaceutical active ingredient in dissolved or lyophilized form.

The present invention relates to compounds of formula I according to claims 1-8 as well as pharmaceutically acceptable salts, tautomers and stereoisomers thereof, including mixtures thereof in all ratios, for use in the treatment of tumors, tumor metastases, proliferative diseases of mesangial cells, hemangiomas, proliferative retinopathies, rheumatoid arthritis, atherosclerotic neovascularization, psoriasis, ocular neovascularization, osteoporosis, diabetes and obesity, lymphoid leukemia, lymphoma, malaria and prostatic hypertrophy.

Use of

The compounds are suitable as pharmaceutically active ingredients for the treatment and control of diseases in mammals, especially humans. These diseases include tumor cell proliferation, pathological neovascularization (or angiogenesis) that promotes the growth of solid tumors, neovascularization in the eye (diabetic retinopathy, age-induced macular degeneration, etc.), and inflammation (psoriasis, rheumatoid arthritis, etc.), and proliferative diseases of mesangial cells.

The invention comprises the use of compounds of formula I and/or physiologically acceptable salts and solvates thereof for the preparation of a medicament for the treatment or prevention of tumors, neoplastic diseases and/or tumor metastases.

The neoplastic disease is preferably selected from the group consisting of tumors in the following regions: squamous epithelium, bladder, stomach, kidney, head and neck, esophagus, cervix, thyroid, intestine, liver, brain, prostate, genitourinary tract, lymphatic system, stomach, larynx, lung, skin, monocytic leukemia, lung adenocarcinoma, small cell lung cancer, pancreatic cancer, glioblastoma, breast cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoid leukemia, chronic lymphoid leukemia, hodgkin lymphoma, non-hodgkin lymphoma.

Also included is the use of a compound according to claim 1 and/or physiologically acceptable salts and solvates thereof according to the present invention for the preparation of a medicament for the treatment of osteoporosis, diabetes and obesity.

Also included is the use of a compound according to claim 1 and/or physiologically acceptable salts and solvates thereof according to the invention for the preparation of a medicament for the treatment or prevention of diseases in which angiogenesis is involved.

Diseases of this type involving angiogenesis are ocular diseases such as retinal angiogenesis, diabetic retinopathy, age-induced macular degeneration, and the like.

The angiogenic disease is preferably selected from diabetic retinopathy, arthritis, cancer, psoriasis, kaposi's sarcoma, hemangioma, myocardial angiogenesis, atherosclerotic plaque neovascularization, angiogenic ocular disease, choroidal neovascularization, retrolental fibroplasia, macular degeneration, corneal graft rejection, rubeosis iridis, neuromuscular glaucoma (neurosculares Glaukom), Oster Webber syndrome.

The proliferative disease of mesangial cells is preferably selected from glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes, transplant rejection, glomerulopathy.

The use of compounds of formula I and/or physiologically acceptable salts and solvates thereof for the preparation of a medicament for the treatment or prevention of inflammatory diseases also falls 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.

The inflammatory disease is preferably selected from inflammatory bowel disease, arthritis, atherosclerosis, asthma, allergy, inflammatory kidney disease, multiple sclerosis, chronic obstructive pulmonary disease, inflammatory skin disease, periodontal disease (Pardontalerkrankung), psoriasis, T-cell mediated immune disease.

The inflammatory bowel disease is preferably selected from ulcerative colitis, Crohn's disease, non-specific colitis.

The T-cell mediated immunological disorder is preferably selected from the group consisting of allergic encephalomyelitis, allergic neuritis, transplant rejection, graft-versus-host reaction, myocarditis, thyroiditis, nephritis, systemic lupus erythematosus, and insulin-dependent diabetes.

The arthritic condition is preferably selected from the group consisting of rheumatoid arthritis, osteoarthritis, kaplan syndrome, feldian syndrome, sjogren's syndrome, ankylosing spondylitis, still's disease, chondrocalcinosis, metabolic arthritis, rheumatic fever, reiter's disease, Wissler's syndrome.

The inflammatory renal disease is preferably selected from glomerulonephritis, glomerular injury, nephrotic syndrome, interstitial nephritis, lupus nephritis, hemorrhagic pulmonary nephritis syndrome, wegener's granulomatosis, renovascular inflammation, IgA nephropathy, idiopathic glomerular disease.

The inflammatory skin disease is preferably selected from psoriasis, atopic dermatitis, contact sensitivity, acne.

Also included is the use of a compound of formula I and/or physiologically acceptable salts and solvates thereof for the preparation of a medicament for the treatment or prevention of a disease or condition in a mammal, in which method a therapeutically effective amount of a compound according to the invention is administered to a diseased mammal in need of such treatment. The amount of treatment depends on the particular disease and can be determined by one skilled in the art without undue effort.

The invention also comprises the use of a compound of formula I and/or physiologically acceptable salts and solvates thereof for the preparation of a medicament for the treatment or prevention of retinal angiogenesis.

Also included is the use of a compound of formula I and/or a physiologically acceptable salt thereof for the manufacture of a medicament for the treatment and/or combating of diseases caused by tumours in mammals, in which method a therapeutically effective amount of a compound of the invention is administered to a sick mammal in need of such treatment. The amount of treatment depends on the particular disease and can be determined by one skilled in the art without undue effort.

The disclosed compounds of formula I may be administered in combination with other therapeutic agents, including anticancer agents. As used herein, the term "anti-cancer agent" relates to any agent that is administered to a cancer patient for the purpose of treating cancer.

The compounds of formula I may also be administered with other well-known therapeutic agents selected for their particular suitability for the condition being treated.

The present compounds are also suitable for combination with known anticancer agents. These known anti-cancer agents include the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, isoprene-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and other angiogenesis inhibitors. The compounds are particularly suitable for co-administration with radiotherapy. "Estrogen receptor modulator" refers to a compound that interferes with or inhibits the binding of estrogen to the receptor, and does not depend on the mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, indoxyl, LY353381, LY 117081, toremifene, fulvestrant, 4- [7- (2, 2-dimethyl-1-oxopropoxy-4-methyl-2- [4- [2- (1-piperidinyl) -ethoxy ] phenyl ] -2H-1-benzopyran-3-yl ] phenyl-2, 2-dimethyl-propionate, 4' -dihydroxybenzophenone-2, 4-dinitrophenylhydrazone, and SH 646.

"androgen receptor modulators" refers to compounds that interfere with or inhibit the binding of androgens to the receptor, and does not depend on any mechanism. Examples of androgen receptor modulators include finasteride and other 5 α -reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole, and abiraterone acetate.

"retinoid receptor modulators" refers to compounds that interfere with or inhibit the binding of retinoids to the receptor, and does not depend on any mechanism. Examples of such retinoid receptor modulators include bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, alpha-difluoro-methyl ornithine, ILX23-7553, trans-N- (4' -hydroxyphenyl) tretinoamide (retinamid) and N-4-carboxyphenyl-tretinoamide.

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

Examples of cytotoxic agents include Tirapazimin (Tirapazimin), Sertenef, cachectin, ifosfamide, tasolinamine, lonidamine, carboplatin, altretamine, thelenemetine, dibromodulcitol, ramustine, fotemustine, nedaplatin, oxaliplatin, temozolomide, Heptaplatin (Heptaplatin), estramustine, enpronaphthalene tosylate, trofosfamide, nimustine, dibromospiro ammonium chloride, meperimidine, lobaplatin, satraplatin, methyl mitomycin (Profiromycin), cisplatin, ilobufen, dexifosfamide (Dexifosfamid), cis-amine dichloro (2-methylpyridine) platinum, benzylguanine, glufosfamide, GPX100, (trans ) -bis-mu- (hex-1, 6-diamine) platinum (II) ] -bis (tetrachloro-platinum (II) diamine (tetrachloro-bis (II) ] -platinum (tetrachloro-bis (platinum) diamine (II) bis (tetrachloro-platinum (II) ] Diarizidinyl-sper min, arsenic trioxide, 1- (11-dodecylamino-10-hydroxyundecyl) -3, 7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, naproxen-4, valrubicin, amrubicin, antineoplastic, 3 '-deamino-3' -morpholino-13-deoxy-10-hydroxycarminomycin, liposomal anthracycline (Annamycin), calicheacin, irinotecan, MEN10755 and 4-desmethoxy-3-desamino-3-aziridinyl-4-methylsulfonyl daunorubicin (see WO 00/50032), but are not limited thereto.

Examples of tubulin inhibitors include paclitaxel, vindesine sulfate, 3',4' -didehydro-4 '-deoxy-8' -nonvincalaukobastine, docetaxel, rhizomycin, Dolastatin (Dolastatin), mevibulin isethionate (Mivobulin-isethionat), Auristatin, cimadrol, RPR109881, BMS184476, vinflunine, Cryptophycin, 2,3,4,5, 6-pentafluoro-N- (3-fluoro-4-methoxyphenyl) benzenesulfonamide, anhydrovinblastine, N-dimethyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-tert-butylamide, TDX258 and BMS 188797.

Topoisomerase inhibitors are, for example, topotecan, Hycaptamin, irinotecan, rubitecan, 6-ethoxypropionyl-3 ',4' -O-exo-benzylidene-churamicin, 9-methoxy-N, N-dimethyl-5-nitropyrazolo [3,4,5-kl ] acridine-2- (6H) propylamine, 1-amino-9-ethyl-5-fluoro-2, 3-dihydro-9-hydroxy-4-methyl-1H, 12H-benzo [ de ] pyrano [3',4': b,7] indolino [1,2b ] quinoline-10, 13(9H,15H) -dione, lurtotecan, 7- [2- (N-isopropylamino) ethyl ] - (20S) camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sorchesen, 2 '-dimethylamino-2' -deoxy-etoposide, GL331, N- [2- (dimethyl-amino) -ethyl ] -9-hydroxy-5, 6-dimethyl-6H-pyrido [4,3-b ] carbazole-1-carboxamide, Asulacrin, (5a,5aB,8aa,9b) -9- [2- [ N- [2- (dimethylamino) ethyl ] -N-methylamino ] ethyl ] -5- [ 4-hydroxy-3, 5-dimethoxyphenyl ] -5,5a,6,8,8a, 9-hexahydrofuro (3',4':6,7) naphtho (2,3-d) -1, 3-dioxol-6-one, 2,3- (methylenedioxy) -5-methyl-7-hydroxy-8-methoxy-benzo [ c ] phenanthridinium, 6, 9-bis [ (2-amino-ethyl) amino ] benzo [ g ] isoquinoline-5, 10-dione, 5- (3-amino-propyl-amino) -7, 10-dihydroxy-2- (2-hydroxyethyl-amino-methyl) -6H-pyrazolo [4,5,1-de ] -acridin-6-one, N- [1- [2 (diethylamino) -ethylamino ] -7-methoxy-9-oxo-9H-thioxanthen-4-yl-methyl ] carboxamide, N- (2- (dimethyl-amino) ethyl) acridine-4-carboxamide, 6- [ [2- (dimethylamino) -ethyl ] -amino ] -3-hydroxy-7H-indeno [2,1-c ] -quinolin-7-one and desmedisodium.

"antiproliferative agents" include antisense RNA and DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231 and INX3001 and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimethadrine, fludarabine, capecitabine, galocitabine-octaalkyl sodium phosphate (cytarabine-ochfosfat), Fosteabin-Natriumhydrat, raltitrexed, Patrexed, ethirimol, thiazolyline, decitabine, noralatrexed, pemetrexed, nelarabine, 2' -deoxy-2 ' -methylenecytidine, 2' -fluoromethylene-2 ' -deoxycytidine, N- [5- (2, 3-dihydro-benzofuranyl) sulfonyl ] -N ' - (3, 4-dichlorophenyl) urea, N6- [ 4-N- [2- (2), 4(E) -tetradecadienoyl ] glycyl-amino-L-glycero-B-L-mannose-heptopyranosyl (mano-pyranosyl) ] adenine, aplidine, ecteinascidin, troxacitabine, 4- [ 2-amino-4-oxo-4, 6,7, 8-tetrahydro-3H-pyrimido [5,4-B ] [1,4] thiazin-6-yl- (S) -ethyl ] -2, 5-thenoyl-L-glutamic acid, aminopterin, 5-fluorouracil, alafenacin, 11-acetyl-8- (carbamoyloxymethyl) -4-formyl-6-methoxy-14-oxa-1, 11-diazacyclo (7.4.1.0.0) -tetradecane-2, 4, 6-trien-9-yl acetate, swainsonine, lometrexol, dexrazine, methioninase, 2 '-cyano-2' -deoxy-N4-palmitoyl-1-B-D-arabinofuranosyl cytosine, and 3-aminopyridine-2-carboxaldehyde thiosemicarbazone. "antiproliferative agents" also include anti-growth factor monoclonal antibodies other than those listed under "angiogenesis inhibitors", such as trastuzumab, and tumor suppressor genes delivered by recombinant virus-mediated gene transfer, such as p53 (see, e.g., U.S. patent No. 6,069,134).

In vitro demonstration of the Effect of pharmacological inhibitors on the proliferation/Activity of tumor cells

1.0 Background

In the present experimental description, inhibition of tumor cell proliferation/tumor cell viability by active ingredients is described.

Cells were seeded in microtiter plates (96-well format) at the appropriate cell density and test substances were added in concentration series. After further 4 days of culture in serum-containing medium, the tumor cell proliferation/tumor cell viability can be determined by means of the alamar blue test system.

2.0 Experimental methods

2.1 Cell culture

For example, commercially available colon cancer cell lines, ovarian cell lines, prostate cell lines, or breast cell lines, and the like.

The cells are cultured in a medium. At intervals of several days, the cells were detached from the culture dish by means of a trypsin solution and seeded in fresh medium at suitable dilutions. At 37 ℃ and 10% CO₂Cells were cultured under the conditions described above.

2.2. Cell seeding

A defined number of cells (e.g. 2000 cells) are seeded in a microtiter plate (96-well cell culture plate) with a multichannel pipettor per well in a volume of 180 microliters of medium. The cells were then cultured in a CO2 incubator (37 ℃ C. and 10% CO 2).

2.3. Addition of test substances

The test substances are dissolved, for example, in DMSO and then used in the cell culture medium at the corresponding concentrations (if necessary, in dilution series). The dilution scale can be adjusted according to the potency of the active ingredient and the desired spread in concentration. The cell culture medium is added to the test substance at the corresponding concentration. The test substance may be added to the cells on the same day as the seeding of the cells. To this end, in each case 20 μ l of substance solution from the pre-dilution plate was added to the culture/well. Cells were incubated at 37 ℃ and 10% CO₂The cells were cultured for 4 days.

2.4. Measurement of color reaction

Add 20. mu.l AlamarBlue reagent to each well and incubate the microtiter plates in a CO2 incubator (at 37 ℃ and 10% CO 2) for an additional 7 hours, for example. The plates were measured at a wavelength of 540 nm in a reader with a fluorescence filter. The plate can be shaken gently just before the measurement.

3. Evaluation of

The absorbance values of the media control (without cells and test substance) were subtracted from all other absorbance values. The control (cells without test substance) is set equal to 100% and represents all the other absorbance values relative to it (for example in% of the control):

and (3) calculating:

determination of IC by statistical programs, e.g. RS1₅₀Value (50% inhibition).

IC of the Compounds according to the invention₅₀The data are shown in table 1.

Determination of the inhibition of proliferation of inhibitors of methionine aminopeptidase 2 in the BrdU proliferation assay (cellular assay)

Proliferation inhibition was determined by adding bromodeoxyuridine (BrdU) to human umbilical vein endothelial cells (HUVEC, Promo-Cell, C-12200). At 37 ℃ and 5% CO₂HUVEC were then cultured in basal medium (Promocell, C-22200) containing supplement cocktail (Promocell, C-39225). After stripping the cells by means of trypsin/EDTA, the number of viable cells was determined and the cells were seeded at a density of 1000 cells per well in a total volume of 175. mu.l (wells previously plated with supplemented medium at 37 ℃ for 1-2 hours or 1.5% gelatin at 37 ℃ for 0.5-2 hours). After 24 hours of culture, test substances are added at various concentrations (e.g., in 10-fold stepwise dilutions, final concentrations of 30 μ M to 0.03 nM) and in volumes of 25 microliters. The DMSO concentration was kept constant at 0.3%. After a total of 48 or 72 hours of culture, 20 microliters of bromodeoxyuridine (Roche, #11647229001, 1:1000 dilution in medium, final concentration 10. mu.M) was added and cultured for another 20 to 24 hours. After a total of 72 or 96 hours of incubation with the test substance, the medium was removed and an immunohistochemical assay was performed to detect BrdU incorporation (BrdU ELISA, Roche, # 11647229001). For this, cells were treated with fixative for 30 minutes at room temperature, followed by incubation with peroxidase-labeled anti-BrdU antibody (diluted 1:100 in antibody dilution buffer) for 60 minutes at room temperature. After three washes with 1-fold concentrated DPBS buffer (Gibco, # 14200), the enzymatic reaction was initiated in TMB substrate solution. After 15 minutes the color development was stopped by adding 25. mu.l of 1M sulfuric acid solution. Optical density measurements were performed by measuring at 450 nM wavelength over 5 minutes. The controls used were wells or empty wells (blank values) containing DMSO-treated cells (100% control). The sensitivity of this test to methionine aminopeptidase inhibitors was examined and confirmed by using the inhibitor fumagillin.

MetAP-2 Activity measurement

By passingCoupling enzyme reaction is used for determining the activity of MetAP-2. The tripeptide Met-Arg-Ser (MAS) is used as substrate. The methionine released is first converted to Met by L-amino oxidase (AAO)_oxAnd H₂O₂. In a second step, with the aid of H₂O₂Peroxidase (POD) catalyzes the formation of o-dianisidine from the leuco dye o-dianisidine_oxThe increase was detected photometrically at 450 nm.

The MetAP-2 activity can be continuously recorded as kinetics. The reaction scheme shows that 1 mole of o-dianisidine is formed per mole of methionine_ox. The MetAP-2 enzyme activity can therefore be directly calculated as delta absorption per time unit. Can be assisted by o-dianisidine_oxExtinction coefficients MetAP-2 activity (moles Met/time unit) was assessed. The change in extinction per time unit is plotted and slope calculations are made in the visually linear region of the reaction.

The activity of this compound is summarized in table 1.

Solubility measurement

Determination by "Shake flask solubility measurement

Eluent preparation:

eluent A of 2 ml diethylamine for synthesis +

1000 ml of methanol, LiChrosolv

Eluent B5 g ammonium acetate for synthesis +

5 ml of methanol, LiChrosolv +

995 ml ultrapure water

Sample solvent:

buffer solution: 3.954 g of sodium dihydrogen phosphate monohydrate + 6.024 g of sodium chloride + 950 ml of ultrapure water, the pH was adjusted using 0.1M NaOH or 0.1M HCl.

Sample preparation:

the sample was shaken at 37 ℃ and 450 rpm for 24 hours.

After about 7 hours, the pH of the sample was checked and, if necessary, adjusted.

The sample was also checked for the presence of excess.

The samples were again checked for pH and sediment just before the end of the 24 hour shaking period.

Ultrapure Water device MilliQ gradient, MilliPore, Instrument F3PN37462D

Shaker TiMix control, Buhler

Incubation hood TH 15 Buhler

766 Calimatic Knick instrument pH 1

pH electrode InLab 423 Mettler

APCI-MS (atmospheric pressure chemical ionization-mass spectrometry) (M + H)⁺。

The racemic end products or racemic intermediates of the compounds according to the invention can be separated simply and on an analytical scale and on a preparative scale by means of chiral HPLC or SFC columns.

§

NMR spectra after addition of deuterated trifluoroacetic acid

The method comprises the following steps:

%

HPLC-MS analysis conditions:

1. column Acquity BEH C-18 (2.1X 100mm, 1.7 μm)

2. Mobile phase A-5 mM ammonium acetate in water, B-acetonitrile

3. Flow mode gradient

4. The flow rate was 0.3 ml/min.

5. UV maximum 254.0 nm

6. Column temperature 30.0 degree

7. Sample preparation acetonitrile + Water

HPLC La Chrom device

Chromolite Performance RP18-e100-4.6mm

Gradient acetonitrile comprising 0.01% formic acid_H2O

Method monolith/monolith (expansion)

Flow rate of 3ml/min

$

Column XBridge C8, 3.5 μm, 4.6x 50 mm;

solvent A water + 0.1% of TFA;

solvent B acetonitrile + 0.1% of TFA;

the flow rate is 2 ml/min;

gradient of 0 min: 5% B, 8min: 100% B, 8.1 min: 10%;

detection at 254 nm

1)

Separation of enantiomers:

separation was performed on Chiralcel OD-H with n-heptane/ethanol = 70/30.

The material was dissolved in 10 ml of n-heptane/EtOH = 1/1 and separated by a 5x25cm Chiralcel OD column containing 20 micron material at a flow rate of 100ml/min of n-heptane/ethanol = 70/30.

In this context, all temperatures are expressed in degrees Celsius. In the following examples, "conventional work-up" means: if necessary, water is added, the pH is adjusted, if necessary, to a value between 2 and 10, depending on the composition of the end product, extraction is carried out with ethyl acetate or dichloromethane, separation, drying of the organic phase over sodium sulfate, evaporation and purification of the product by chromatography on silica gel and/or by crystallization.

F. Melting point

Mass Spectrometry (MS) EI (Electron impact ionization) M⁺

FAB (fast atom bombardment) (M + H)⁺

ESI (electrospray ionization) (M + H)⁺

Synthetic schemes for preparing compounds of formula I:

a)

b)

c)

a) route 1: the acid is converted to the corresponding amide in an amide coupling and then oxidized at the carbon between the two carboxyl groups using t-butyl hydroperoxide in an alkaline environment to yield the alcohol (followed by separation of the enantiomers by chromatography).

b) Route 2: alternatively, it has been found that the ethyl ester of the starting acid cannot be oxidized under the same oxidation conditions as described in route 1 for the amide. However, this can be achieved with cerium chloride. In this case, the chlorine derivative obtained at 30% can be separated into enantiomers similarly to the alcohol, and then further derivatized. (no further reaction is shown here, but the products are then for example "A39", "A43" and "A61").

c) However, the use of cerium fluoride does not produce similar fluorine derivatives. Reaction of an alcohol compound with DAST (diethylaminosulfur trifluoride) in dichloromethane produces the desired fluorine analog. Tests confirmed the inversion at the asymmetric center expected from the principle of using DAST fluorinated alcohols.

d)

In d) it is shown how N-aryllactam carboxylic acids can be prepared. This route can be easily carried out if the nitrogen compound is a liquid. It was found that if the nitrogen-aryl compound is not a liquid, co-melting of the starting materials is not advantageous. However, if the nitrogen-aryl compound is melted and a dicarboxylate (Meldrum's acid) is added to this solution, the desired product is obtained.

e)

The route for preparing N-alkylaryl lactam carboxylic acids is shown in e).

f)

In f) is shown another alternative to proceed with making available the aryl group on the lactam nitrogen, where the analogous nitrogen compound is not a liquid. Here, the lactam-nitrogen is coupled to the aryl halide, this step preferably being catalyzed by transition metal compounds.

Examples 1

Synthesis of 3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (2-thiophen-2-yl-ethyl) amide ("a65") [ analogous to synthesis scheme a); route 1)

a) 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (2-thiophen-2-yl-ethyl) amide

200 mg of 1-phenyl-2-oxo-3-pyrrolidinecarboxylic acid, 155 mg of 2-thienylethylamine hydrochloride, 550 mg of N- (3-dimethylaminopyrrolyl) -N' -ethyl-carbodiimide hydrochloride, 190 mg of hydroxy-benzotriazole, 132 microliters of triethylamine and 5 ml of DMF are combined in a microwave container. After sealing the container with a septum, the mixture was heated (160 ℃,5 min) by means of microwaves (Emrys optimier, personal chemistry). Water was added to the batch. A precipitate formed. Water was added until no more precipitate formed. The precipitate is filtered off with suction and dried.

Yield: 239 mg of (A);

appearance: a white solid;

purity: 100% (220nm, UV tracing by LC-MS);

LC-MS: 315(M+H)；

HPLC: 3.03 (Rt/min)；

HPLC method 1-100-2 (Instrument: LaChrom)

Column Chromolith Performance RP18e 100-3mm

Flow rate 2 ml/min (Pump: L-7100)

Solvent A, water + 0.05% HCOOH

Solvent B acetonitrile + 0.04% HCOOH

Wavelength 220nm (detector: L-7455)

Gradient of 99% A in 0-0.2 min, 99% A in 0.2-3.8 min, 100% B in 3.8-4.4 min, 100% B in 4.4-4.5 min, 99% A in 4.5-5.1 min;

LC-MS method (Instrument Agilent 1100 series)

Column Chromolith Speed RodRP18e-50-4.6

Flow rate of 2.4ml/min

Solvent A, water + 0.05% HCOOH

Solvent B acetonitrile + 0.04% HCOOH

Wavelength of 220nm

Gradient from 4% B to 100% B in 0-2.8min and 100% B in 2.8-3.3 min.

b) 3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (2-thiophen-2-yl-ethyl) amide ("A65")

100 mg of the amide prepared under a) are dissolved in 10 ml of tert-butanol and 60 mg of tert-butyl hydroperoxide (70% in water) and 0.2 ml of sodium ethoxide (20% in ethanol) are added, and the mixture is stirred at 90 ℃ for 4 hours. The batch was concentrated almost to dryness. Water and EE (ethyl acetate) were added to the residue and shaken. The organic phase is passed through Na₂SO₄Dried, filtered off with suction and concentrated to dryness in vacuo: 80 mg were obtained. The residue was dissolved in DMSO and purified by preparative HPLC. The product fractions were combined and concentrated to dryness in vacuo. 57 mg (54%) "A65" was obtained;

appearance: a white solid;

purity: 100% (220nm, UV tracing by LC-MS);

LC-MS: 331 (M+H)；

HPLC: 2.88 (Rt/min)；

HPLC method 1-100-2 (Instrument: LaChrom)

Column Chromolith Performance RP18e 100-3mm

Flow rate 2 ml/min (Pump: L-7100)

Solvent A, water + 0.05% HCOOH

Solvent B acetonitrile + 0.04% HCOOH

Wavelength 220nm (detector: L-7455)

LC-MS method (Instrument Agilent 1100 series)

Column Chromolith Speed RodRP18e-50-4.6

Flow rate of 2.4ml/min

Solvent A, water + 0.05% HCOOH

Solvent B acetonitrile + 0.04% HCOOH

Wavelength: 220nm

Gradient from 4% B to 100% B in 0-2.8min and 100% B in 2.8-3.3 min;

preparative HPLC method 15-35-10-50 ml _ empfind _ o _ equi.M (Instrument: Agilent 1100 series)

Column Chromolith Prep RodRP18e

Flow rate of 50ml/min

Solvent A acetonitrile + 0.1% TFA

Solvent B Water + 0.1% TFA

Wavelength: 220nm

Gradient from 1-15% ACN in 2 min over 2 min, from 15-35% ACN over 8min, collected from 2 min to 11 min.

Separation of the racemate into the enantiomers:

the separation was analyzed on a Chiralcel OD-H with n-heptane/ethanol = 70: 30.

40 mg of racemate were dissolved in 10 ml of n-heptane/EtOH = 1:1 and separated by 5x25cm Chiralcel OD column containing 20 micron material at 100ml/min n-heptane/ethanol = 70/30 flow rate.

2 fractions were collected and concentrated.

Fraction 1: m =20 mg enantiomer 1:

pure enantiomers

Fraction 2: m =20 mg enantiomer 2:

ratio of enantiomers:

enantiomer 1: 0.6% and enantiomer 2: 99.4%.

Examples 2

Preparation of 3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid [ synthesis scheme b) intermediate; route 2)

a)

Thionyl chloride (5.8 g) was added dropwise to a solution of 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (5 g) in 5 ml ethanol at 0 ℃ and stirred for 4 hours at 80 ℃.

After complete reaction, the solvent was removed in vacuo and the residue was partitioned between water and ethyl acetate. The organic phase is washed with saturated sodium bicarbonate solution and saturated sodium chloride solution and dried over sodium sulfate. After removal of the solvent and the drying agent, pure 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester is obtained which is sufficient for further reaction;

LCMS mass found (M +1, 234);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5u) + ve mode;

rt (min) 3.360 area% 98.577.

b)

Cerium chloride heptahydrate (1.59 g) was added to a solution of 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester (5 g) in 10 ml isopropanol and O was passed over₂Gas was allowed to flow for 15 minutes, and then the mixture was allowed to stir for 14 hours.

All volatile constituents are then removed in vacuo and the batch is purified by chromatography.

The resulting 3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester was analyzed using chiral HPLC and subsequently separated into the enantiomers.

The 'R' enantiomer elutes after rt (min) 8.197.

The 'S' enantiomer elutes after rt (min) 11.240;

(Chiralpak AD-H (250x4.6) mm, 5 μm; n-hexane/IPA = 60: 40).

LCMS mass found (M +1, 250);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5u) + ve mode;

rt (min) 2.973 area% 98.698.

c)

Reacting LiOH/H₂O (750 mg) was added to (S) -3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester (1.5 g) in THF/H₂O = 3:1, and the mixture is stirred for 4 hours.

The solvent was then removed in vacuo and the batch was acidified using 1.5N HCl. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried over sodium sulfate. Obtaining (S) -3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid;

LCMS mass found (M +1, 222);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5u) + ve mode;

rt (min) 1.970 area% 98.63;

chiral HPLC:

the method comprises the steps of mixing normal hexane and IPA 60: 40; the flow rate is-1.0 ml/min;

a column of ChiralpakAD-H (250X4.6) mm, 5 μm;

rt (min) 9.794 area% 98.698.

Amide coupling

EDCl (3 equiv.) and HOBT (1.5 equiv.) are added to a solution of the acid (1 equiv.) and amine (1.2 equiv.) prepared above in dry DMF (3 mml/mmol) and the mixture is heated in a microwave for 20 minutes at 160 ℃.

When the reaction was complete, the solvent was removed in vacuo and purified by chromatography.

Examples 3

Preparation of (S) -1- (3-chlorophenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid (2-thiophen-2-yl-ethyl) amide ("A37")

a)

3-Chloroanilide (10.45 g) was added to cyclopropyl-Meldrum's acid (6, 6-dimethyl-5, 7-dioxaspiro [2.5] octane-4, 8-dione; 7 g) in dry dichloromethane and the mixture was stirred at RT for 14 h.

The batch was then washed with 10% NaOH solution and the aqueous phase was acidified with 1N HCl until a colorless precipitate formed. This was filtered off and dried in vacuo;

LCMS mass found (M +1, 240);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ) + ve mode;

rt (min) 3.073 area percent 98.91.

b)

The acid prepared under a) (0.2 g) was dissolved in anhydrous dichloromethane. 2-Thiophenylethylamine (127 mg), triethylamine (25 mg) and T3P (399 mg) were then added and the batch was stirred for 14 hours. The mixture was subjected to an aqueous workup and extracted with dichloromethane.

The organic phase is washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate, evaporated and chromatographed on silica gel;

LCMS mass found (M +1, 349);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ) + ve mode;

rt (min) 4.417 area% 99.65.

c)

The amide prepared under b) was dissolved in tert-butanol (100 mg). A solution of tert-butyl hydroperoxide (70% equiv.) (0.051 g) and sodium ethoxide (20% in ethanol; 39 mg) were added and the mixture was stirred at 70 ℃ for 1 hour.

After completion of the reaction, the solvent was removed in vacuo and water was added until a colorless precipitate formed. This was filtered off and chromatographed.

The enantiomers were obtained by chiral preparative HPLC.

The enantiomer eluting after rt (min) 5.419 was compound "a37" in active S-configuration; (Chiralpak AD-H (250x4.6) mm, 5 μm; n-hexane/IPA = 60: 40);

LCMS mass found (M +1, 365);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ) + ve mode;

rt (min) 4.055 area% 99.02;

HPLC>99.25%:

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ);

Rt (min): 4.102；

chiral HPLC:

the method comprises the steps of (1) adding normal hexane and IPA = 60: 40; flow rate-1.0 ml/min

A column of Chiralpak AD-H (250X4.6) mm, 5 μm;

rt (min) 5.419 area% 100.

Examples 4

Preparation of (S) -3-fluoro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (2-thiophen-2-yl-ethyl) amide ("A86

a)

300 mg of (R) -3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester were dissolved in anhydrous dichloromethane, 244 mg of DAST were slowly added at-78 ℃ and the mixture was stirred for a further 4 hours.

After completion of the reaction, the batch was poured into saturated sodium bicarbonate solution, the aqueous phase was extracted with dichloromethane and the combined organic phases were dried over sodium sulfate. The drying agent and solvent were removed and the residue (180 mg) was further reacted in a subsequent reaction without further purification.

LCMS mass found (M +1, 252);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5u) + ve mode;

rt (min) 3.745 area% 93.764.

b)

84 mg of LiOH/H₂O is added to 180 mg of (S) -3-fluoro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acidEthyl ester in THF/H₂O = 3:1 and the mixture is stirred at RT for 4 hours. After the reaction was complete, the batch was neutralized using 1.5N HCl. The aqueous phase was extracted with ethyl acetate and the organic phase was dried over sodium sulfate. The solvent was removed in vacuo to give 90 mg of (S) -3-fluoro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid;

LCMS mass found (M +1, 222);

the method comprises the steps of A-0.1% HCOOH and B-MeOH, wherein the flow rate is =1.0 ml/min;

atlantis DC18(50x4.6mm.5 mu) + ve mode;

rt (min) 1.948 area% 90.84.

c)

230 mg of EDCl, 65 mg of HOBt are added to a solution of 90 mg of (S) -3-fluoro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid and 61 mg of 2-thiophen-2-yl-ethylamine in anhydrous DMF and the mixture is heated in microwave at 160 ℃ for 20 min. After completion of the reaction, all volatile components were removed in vacuo and the residue was purified by chromatography. 23 mg of "A86" were obtained;

LCMS mass found (M +1, 333);

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow rate-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ) + ve mode;

rt (min) 4.147 area% 97.65;

HPLC>97%:

method A-in H₂0.1% TFA in O, B-0.1% TFA in acetonitrile flow-2.0 ml/min;

column X Bridge C8(50x4.6mm.3.5 μ);

Rt (min) : 4.209。

examples 5

Preparation of (S) -3-amino-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (3-chloro-5-fluorobenzyl) amide ("A91a")

Examples 6

Synthesis of 6.12-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester

Thionyl chloride (5.8 g) was added to a solution of 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (5 g) in ethanol at 0 ℃ and the batch was stirred at 80 ℃ for 4 hours. After completion of the reaction, the batch was concentrated in vacuo, the residue taken up in water and extracted with ethyl acetate. The organic phase is washed with saturated sodium bicarbonate solution and saturated sodium chloride solution and dried over sodium sulfate. Obtaining 2-oxo-1-phenyl-pyrrolidine-3-ethyl formate (4.5 g, 79%);

HPLC:

method A-in H₂0.1% TFA in O, B-0.1% TFA in ACN flow-2.0 ml/min.

Column X Bridge C8(50x4.6mm.3.5u) + ve mode

Rt (min) 3.360 area% 98.577.

Synthesis of 2 (S) -and (R) -3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester and 3-chloro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester

Cerium (III) chloride heptahydrate (1.59 g) was added to a solution of 2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester (5 g) in isopropanol and O was passed through₂For 15 minutes. The batch is stirred at RT for 12 hours. After completion of the reaction, the solvent was removed in vacuo and the residue was purified by chromatography on silica gel. 3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester (2.7 g) was analyzed by chiral HPLC and purified by this method. The material eluting at rt (min) 8.197 is the "R" enantiomer (1.2 g) and the material eluting at rt (min) 11.240 is the "S" enantiomer (1.2 g);

(Chiralpak AD-H (250x4.6) mm, 5 μm; n-hexane/isopropanol = 60: 40).

The desired S enantiomer is further used for the synthesis of alcohol derivatives.

In this oxidation 3-chloro-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester was also formed as racemate in 20% yield. This was isolated by chromatography on silica gel (1 g) before the enantiomeric separation of the alcohol and was also used in the next step in racemic form.

6.3 Synthesis of Ethyl 33-azido-2-oxo-1-phenylpyrrolidine-3-carboxylate

Adding NaN₃(58 mg, 0.89 mmol) was added to a solution of ethyl 3-chloro-2-oxo-1-phenylpyrrolidine-3-carboxylate (0.2 g, 0.74 mmol) in dry DME (5 mL). The reaction solution was stirred at 75 ℃ for 14 hours under nitrogen. After complete reaction, the batch is diluted with ethyl acetate and extracted with water. The organic phase was dried over sodium sulfate, concentrated and the residue was directly used on to the next step (200 mg, 97%);

thin layer chromatography Hexane/Ethyl acetate = 7: 3R_f: 0.4；

。

6.6 Synthesis of 43-azido-2-oxo-1-phenylpyrrolidine-3-carboxylic acid

LiOH (91 mg, 2.1 mmol) was added to 3-azido-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid ethyl ester (0.2 g, 0.7 mmol) in THF (2 ml) and H₂O (1 ml) and the batch was stirred at RT for 20 h. After complete reaction, all volatile components were removed in vacuo and the residue was diluted with water and acidified with 1.5N HCl solution. The resulting precipitate was filtered off and washed with water. Drying in vacuo gave a colorless solid as the product. (150 mg, 84%);

thin layer chromatography Hexane/Ethyl acetate (7/3): R_f: 0.4；

。

Synthesis of 53-azido-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (3-chloro-5-fluorobenzyl) amide ("A150")

3-chloro-5-fluorobenzylamine (97 mg, 0.61 mmol) and Et at 0 deg.C₃N (0.24 ml, 1.8 mmol), followed by T3P (0.6 ml, 1.8 mmol) was added to a solution of 3-azido-2-oxo-1-phenylpyrrolidine-3-carboxylic acid (150 mg, 0.61 mmol) in 5 ml dichloromethane. The reaction mixture was stirred at room temperature under nitrogen for 4 hours. After completion of the reaction, water was added and the mixture was extracted with dichloromethane to exhaustion. The combined organic phases are then washed with water and saturated sodium chloride solution and dried over sodium sulfate. After filtration and evaporation, the residue was purified by chromatography on silica gel. The product was obtained as a colourless solid (150 mg, 63% yield);

thin layer chromatography chloroform/methanol = 9.5:0.5), R_f:0.3

。

Synthesis of 6.63-amino-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (3-chloro-5-fluorobenzyl) amide ("A151")

Adding Et₃N (0.1 ml, 0.75 mmol), followed by propane dithiol (0.08 ml, 0.75 mmol) was added to a solution of 3-azido-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid (3-chloro-5-fluorobenzyl) amide (100 mg, 0.25 mmol) in 2 ml methanol. The batch is stirred at RT for 12 hours. After completion of the reaction, all volatile components were removed in vacuo and the residue was purified on silica gel.

The following compounds were obtained analogously

The following compounds were prepared analogously:

(S) -3-hydroxy-1-isobutyl-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A121")

，

(S) -1- (2-amino-1-methyl-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A122")

，

(S) -1- (2-dimethylamino-1-methyl-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A123")

，

(S) -1- (2-amino-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A124")

，

(S) -1- (2-dimethylamino-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A125")

，

(S) -1- (3-dimethylamino-propyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A126")

，

(S) -3-hydroxy-1- (3-hydroxy-propyl) -2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A127")

，

(S) -3-hydroxy-1- (3-methoxy-propyl) -2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A128")

，

(S) -3-hydroxy-1- (2-methoxy-ethyl) -2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A129")

，

(S) -1- (2-formylamino-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A130")

，

(S) -1- (2-acetylamino-ethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A131")

，

(S) -1- [1- (acetylamino-methyl) -cyclopropyl ] -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A132")

，

(S) -3-hydroxy-2-oxo-1- (2-ureidoethyl) -pyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A133")

，

(S) -1- (1H-benzimidazol-5-yl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A134")

，

(S) -3-hydroxy-2-oxo-1- (2-oxo-2, 3-dihydro-1H-indol-5-yl) -pyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A135")

，

(S) -3-hydroxy-2-oxo-1- ((S) -2,2, 2-trifluoro-1-formylaminomethyl-ethyl) -pyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A136")

，

1- (3-carbamoyl-phenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A137")

，

1- (2-carbamoyl-phenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A138")

，

{2- [ (S) -3- (3-chloro-5-fluorobenzylcarbamoyl) -3-hydroxy-2-oxopyrrolidin-1-yl ] -ethyl } -carbamic acid tert-butyl ester ("A140")

，

1- (3-cyanophenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A141")

，

1- (2-cyanophenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A142"),

1- (4-cyanophenyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A143"),

1- (3-amino-3-oxopropyl) -N- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-2-oxopyrrolidine-3-carboxamide ("A144")

，

N- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-1- [3- (methylamino) -3-oxopropyl ] -2-oxopyrrolidine-3-carboxamide ("A145")

，

1- (4-amino-4-oxobutyl) -N- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-2-oxopyrrolidine-3-carboxamide ("A146"),

n- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-1- [4- (methylamino) -4-oxobutyl ] -2-oxopyrrolidine-3-carboxamide ("A147"),

1- (2-cyanoethyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A148")

,

1- (3-cyanopropyl) -3-hydroxy-2-oxopyrrolidine-3-carboxylic acid 3-chloro-5-fluorobenzylamide ("A149")

3-hydroxy-1- (6-methyl-pyridin-3-yl) -2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A152")

,

3-hydroxy-2-oxopyrrolidine-1, 3-dicarboxylic acid-3- (3-chloro-5-fluorobenzyl-amide) -1-acetamide ("A153")

,

3-hydroxy-2-oxopyrrolidine-1, 3-dicarboxylic acid-3- (3-chloro-5-fluorobenzyl-amide) -1-phenylamide ("A154")

,

3-azido-N- [ (3-chloro-5-fluorophenyl) -methyl ] -2-oxo-1-phenyl-pyrrolidine-3-carboxamide ("A155")

3-hydroxy-2-oxo-1-p-tolyl-pyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzyl-amide ("A156")

,

1-carbamoylmethyl-3-hydroxy-2-oxopyrrolidine-3-carboxylic acid-3-chloro-5-fluorobenzylamide ("A157")

,

(3S) -1- [ (1S) -2-amino-1-methyl-2-oxoethyl ] -N- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-2-oxopyrrolidine-3-carboxamide ("A158")

,

(3S) -1- [ (1R) -2-amino-1-methyl-2-oxoethyl ] -N- [ (3-chloro-5-fluorophenyl) methyl ] -3-hydroxy-2-oxopyrrolidine-3-carboxamide ("A159"),

3-hydroxy-2-oxo-1- (4-fluorophenyl) -pyrrolidine-3-carboxylic acid-3-trifluoro-methyl-5-fluorobenzylamide ("A160"),

3-hydroxy-2-oxo-1-phenyl-pyrrolidine-3-carboxylic acid-3, 5- (bis-trifluoromethyl) -benzylamide ("a 161").

The following compounds were obtained analogously

LCMS:

Process A to H₂0.1% TFA in O, 0.1% TFA in B-ACN flow-2.0 ml/min.

Column X Bridge C8(50x4.6mm.3.5u) + ve mode

$

LC-MS method (Instrument Agilent 1100 series)

Column Chromolith Speed RodRP18e-50-4.6

Flow rate of 2.4ml/min

Solvent A, water + 0.05% HCOOH

Solvent B acetonitrile + 0.04% HCOOH

Wavelength: 220nm

Gradient from 4% B to 100% B in 0-2.8min and 100% B in 2.8-3.3 min.

$$

Method A-10 mM NH₄HCO₃And B-ACN with a flow rate of-1.0 ml/min.

Column X Bridge C8(50x4.6mm.3.5u) -ve mode

HPLC:

Process A to H₂0.1% TFA in O, 0.1% TFA in B-ACN flow-2.0 ml/min.

Column X Bridge C8(50X4.6mm.3.5u) + ve mode

$$$

The flow rate of the isopropanol is-0.8 ml/min.

The running time is 20 min

Column: Chiralpak AD.

TABLE 1

Inhibition of MetAP-2

IC of the Compounds of formula I according to the invention₅₀

。

The following examples relate to pharmaceutical formulations:

example A: injection vial

A solution of 100 g of the active ingredient of the formula I and 5 g of disodium hydrogen phosphate in 3 l of distilled water is adjusted to pH 6.5 using 2N hydrochloric acid, sterile-filtered, filled into injection vials, lyophilised under sterile conditions and sealed under sterile conditions. Each injection vial contained 5 mg of active ingredient.

Example B: suppository

A mixture of 20 g of the active ingredient of the formula I with 100 g of soya lecithin and 1400 g of cocoa butter was melted, poured into moulds and allowed to cool. Each suppository contains 20 mg of active ingredient.

Example C: solutions of

In 940 ml of double distilled water, 1 g of active ingredient of the formula I, 9.38 g of NaH₂PO₄∙ 2 H₂O, 28.48 g Na₂HPO₄∙ 12 H₂A solution was prepared of O and 0.1 gram of benzalkonium chloride. The pH was adjusted to 6.8, the solution was made up to 1 liter and sterilized by radiation. Such solutions may be used in the form of eye drops.

Example D: ointment

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

Example E: tablet formulation

A mixture of 1 kg of active ingredient of the formula I, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is compressed in a conventional manner into tablets in such a way that each tablet contains 10 mg of active ingredient.

Example F: sugar-coated pill

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

Example G: capsule

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

Example H: ampoule (CN)

A solution of 1 kg of the active ingredient of the formula I in 60 l of heavy distilled water is sterile-filtered, filled into ampoules, lyophilised under sterile conditions and sealed under sterile conditions. Each ampoule contains 10 mg of active ingredient.

Claims

1. A compound of the formula and pharmaceutically acceptable salts and optical isomers thereof,and are and

。

2. a medicament comprising at least one compound according to claim 1 and/or pharmaceutically acceptable salts and optical isomers thereof, and optionally excipients and/or auxiliaries.

3. The use of a compound according to claim 1, and pharmaceutically acceptable salts and optical isomers thereof, for the manufacture of medicaments for the treatment of tumors, tumor metastases, proliferative diseases of mesangial cells, hemangiomas, proliferative retinopathies, rheumatoid arthritis, atherosclerotic neovascularization, psoriasis, ocular neovascularization, osteoporosis, diabetes and obesity, lymphoid leukemia, lymphoma, malaria and prostatic hypertrophy.

4. Use according to claim 3, wherein the neoplastic disease is selected from the group consisting of tumors in the following sites: squamous epithelium, bladder, stomach, kidney, head and neck, esophagus, cervix, thyroid, intestine, liver, brain, prostate, genitourinary tract, lymphatic system, stomach, larynx, lung, skin, monocytic leukemia, lung adenocarcinoma, small cell lung cancer, pancreatic cancer, glioblastoma, breast cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoid leukemia, chronic lymphoid leukemia, hodgkin lymphoma, non-hodgkin lymphoma.

5. Use of a therapeutically effective amount of a compound according to claim 1 and/or a physiologically acceptable salt thereof in combination with a compound selected from the group consisting of: 1) estrogen receptor modulators, 2) androgen receptor modulators, 3) retinoid receptor modulators, 4) cytotoxic agents, 5) antiproliferative agents, 6) prenyl-protein transferase inhibitors, 7) HMG-CoA reductase inhibitors, 8) HIV protease inhibitors, 9) reverse transcriptase inhibitors, and 10) other angiogenesis inhibitors.

6. Use of a therapeutically effective amount of a compound according to claim 1 and/or a physiologically acceptable salt thereof in combination with a compound for the manufacture of a medicament for the treatment of tumors: 1) an estrogen receptor modulator, 2) an androgen receptor modulator, 3) retinoid receptor modulator, 4) a cytotoxic agent, 5) an antiproliferative agent, 6) a prenyl-protein transferase inhibitor, 7) an HMG-CoA reductase inhibitor, 8) an HIV protease inhibitor, 9) a reverse transcriptase inhibitor and 10) other angiogenesis inhibitors, wherein the above compounds are combined with radiation therapy.