DE10034627A1 - New cyclic hydrocarbyl substituted oxazolidinone derivatives useful as tumor necrosis factor-alpha inhibitors in treatment of e.g. atherosclerosis, arthritis, Crohn's disease, osteoporosis, cardiac infarction and psoriasis - Google Patents

Abstract

Cyclic hydrocarbyl substituted oxazolidinone derivatives and their salts are new. The cyclic hydrocarbyl substituted oxazolidinone derivatives of formula (I) and their salts: d = cyclic hydrocarbyl, e.g. aryl, (optionally substituted with 1 or more Rd); b = -CH2-; m = 0-4; a = H,hydrocarbyl or saturated, partly unsaturated or aromatic heterocyclyl with 1-3 S, N or O; e = NH2 (optionally mono- or disubstituted), OH (optionally protected), 1-6C alkoxy (optionally monosubstituted with hydrocarbyl, e.g. aryl) or -x(Re3)-C(O)-y(Re1)(Re2); x,y = O or N; Re1, Re2 = H or 1-6C alkyl; or yRe1Re2 = 5-8 membered, saturated or partly unsaturated heterocyclyl with 1-3 S, N and/or O (optionally substituted), when y = N; Re3 = H or hydrocarbyl, especially 1-6C alkyl, when x = N; Rd = CHO, NO2, CX3, ORd1, 1-8C alkanoyl, alkyl (optionally substituted with 1-6C alkyl), preferably 1-6C alkyl or -NRd2Rd3; X = halo; Rd1 = hydrocarbyl (optionally substituted), e.g. aryl or 1-6C alkyl; Rd2, Rd3 = H, hydrocarbyl, 1-8C alkoxycarbonyl or -NRd4Rd5; and Rd4Rd5 = H, hydrocarbyl or 1-8C alkoxycarbonyl. Provided that: Re2 = absent, when y=O; and Re2, Re3 = absent, when x=y=O.

Description

The present invention relates to new aryl-substituted oxazolidinones with cyto kin inhibitory effect, process for their preparation and their use as Medicines, especially for the treatment of tumor necrosis factor (TNF) triggered effects such. B. Atherosclerosis.

Tumor necrosis factor α (TNFα) is a pro-inflammatory cytokine with athero gener effect. Pro-inflammatory cytokines are proteins that are considered inflammatory promotional messenger substances formed after a stimulus in certain tissue cells become.

TNFα is a cytokine that is also used in many pathological inflammatory reactions particularly promotes inflammation progression. In extreme cases, if TNFα is increasingly released into the blood circulation, contributes to TNFα development of life-threatening conditions such as septic shock disseminated intravascular coagulation or cachexia. With locally limited Release of TNFα promotes inflammatory processes that affect the tissue in to which it is formed remain limited.

Recently it has been reported that TNFα is of particular importance in the pro inflammatory processes in the arterial wall. In this together Menhang is the (atherogenic) effect of INFα in promoting arteriosclerosis several studies have been proven. It could be shown that TNFα is increasingly formed in arteriosclerotic tissue.

(T. J. DeGraba, Neurology 49 Suppl 4, S 15-S 19, 1997; M. Kaartinen et al. Circula tion 94, 2787-2792, 1996, X. Lei et al., Atherosclerosis 125 (1996), 81-89) and the Effect of TNFα on cells of the vascular wall (endothelial cells, smooth muscle cells), and in particular the effect of TNFα on cells of the arteriosclerotic vascular wall,  which additionally monocytes, naive macrophages, foam cells and Contains lymphocytes, has been examined and described in many different ways:

TNFα triggers the formation of cell adhesion molecules, especially on endothelial cells from how B. vascular cell adhesion molecule (VCAM), intercellular adhesion mole cule (ICAM-1) or P-selectin, which in turn is another immigration of Monozy macrophages and lymphocytes for the progression of inflammation of the vascular wall (T. J. DeGraba, Neurology 49 Suppl 4, S 15-S 19, 1997; J. L. Barks et al. J. Immunology 159, 4532-4538, 1997, M.F. Iademarco et al., J. Clin. Invest. 95 (1995), 264-271). TNFα triggers the formation of other cytokines in endothelial cells, Monocytes, foam cells and smooth muscle cells. Interleukin 1 (IL- 1) and autocrine TNFα are mentioned here.

TNFα triggers the formation of chemokines such as monocyte chemoattractant protein (MCP-1) and interleukin 8 (IL-8) in the named vascular cells for further Recruitment of monocytes and lymphocytes into the arteriosclerotically altered Vessel wall.

TNFα triggers the formation of growth factors (e.g. platelet derived growth factor) and matrix metalloproteases, which in turn cause inflammatory processes in the vessel Promote the wall (H. Funayama, Cardiovasc. Res. 37, 216-224 (1998); T. B. Rajavashisth, Circulation 99, 3103-3109 (1999).

TNFα is next to the importance for the progression of arteriosclerotic Vascular inflammation (P. T. Kovanen et al., Circulation 94, 2787-2792, 1996) also from crucial in the pathophysiological progression of other ent ignition diseases for which a synthesis inhibition of the proinflammatory Cytokins TNFα also a therapeutically useful intervention for weakening which represents disease. Inflammatory and autoimmune diseases can be treated by TNFα synthesis inhibition because TNFα is causative contributes to the clinical picture are arthritis, rheumatoid arthritis, osteoporosis,  Crohn's disease, inflammatory lung diseases like Adult Respiratory Distress Syndrome (ARDS), graft rejection, reperfusion tissue damage after stroke, heart attack or peripheral vascular occlusion, chronic ent inflammatory fibrotic organ changes such as liver fibrosis, or the generalized Autoimmune disease systemic lupus erythematosus or other forms of Lupus erythematosus and dermal inflammatory diseases such as psoriasis.

From publications EP-A-0 693 491, EP-A-789 026 and EP-A-789 025 Oxazolidinones with antibacterial effects known.

The present invention relates to aryl-substituted oxazolidinones of the general formula (I)

in the
d represents a cyclic hydrocarbon radical, for example an aryl radical, which in turn may be substituted by one or more radicals R ^d in which b represents CH ₂ and
m can assume the values 0 to 4,
where a can be hydrogen, a hydrocarbon radical or a saturated or partially unsaturated or aromatic heterocycle, having 1 to 3 heteroatoms selected from S, N, O;
in which e can stand for amino, mono- or di-substituted amino, hydroxy, (C ₁ -C ₆ ) alkoxy which in turn can be substituted with a hydrocarbon residue, for example an aryl residue, or a hydroxy protecting group;
where e can stand for a group (eI)

wherein
x or y can stand for oxygen or nitrogen, with the provisos
that when x is nitrogen R ^{e3 is} hydrogen or a hydrocarbon radical, preferably (C ₁ -C ₆ ) alkyl,
that when x and y stand for oxygen, R ^e2 and R ^e3 are omitted,
that when y is oxygen, R ^{e2 is} absent,
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₆ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together with y including a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, which in turn can be further substituted;
R ^d can be selected from one of the groups
-COH, -NO ₂ , -CX ₃ , where X is halogen, -OR ^d1 , (C ₁ -C ₈ ) alkanoyl, alkyl, preferably (C ₁ -C ₆ ) alkyl, which in turn can be substituted with Groups from selected from hydroxy,

in which
R ^d1 can represent a hydrocarbon radical, for example an aryl radical or (C ₁ -C ₆ ) alkyl, which in turn can be substituted;
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon radical, (C ₁ -C ₈ ) alkoxycarbonyl or a group

wherein R ^d4 , R ^d5 can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₈ ) alkoxycarbonyl, and their salts;
Physiologically acceptable salts are preferred in the context of the invention.

Physiologically acceptable salts can be salts of the compound according to the invention with inorganic or organic acids. Salts are preferred with inorganic acids such as hydrochloric acid, bromine water Substance acid, phosphoric acid or sulfuric acid, or salts with organic carbon or sulfonic acids such as acetic acid, maleic acid, fumaric acid, apples acid, citric acid, tartaric acid, lactic acid, benzoic acid, or methanesulfonic acid,  Ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid or naphthalenedisulfone acid.

Physiologically acceptable salts can also be metal or ammonium salts compounds of the invention. Alkali metal salts are particularly preferred (e.g. sodium or potassium salts), alkaline earth salts (e.g. magnesium or calcium salts) and ammonium salts derived from ammonia or organic Amines, such as ethylamine, di- or triethylamine, di- or triethanol amine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine, ethylenediamine or 2-phenylethylamine.

The compounds of general formula (I) according to the invention can be found in ver Different stereoisomeric forms occur that are either like picture and mirror image (enantiomers), or which are not like image and mirror image (diastereomers) behavior. The invention relates to both the enantiomers and the diastereomers as well as their respective mixtures. The racemic shapes can be just like that Diastereomers in a known manner into the stereoisomerically uniform constituents separate; for example, separation by chromatography on chiral phases respectively.

The compounds of the invention and their salts can be used in the context of Er invention also exist as solvates. Such connections are meant here under solvates are understood to contain solvents in the crystal in a defined manner. Such compounds can have one or more, typically 1 to 5 equivalents of the solvent by crystallization of the compound from the corresponding Solvent can be obtained. Typical solvates are e.g. B. the hydrates in Contain crystal water.

Hydrocarbon residues can be saturated or unsaturated, linear, branched or be cyclic and have 1 to 24 carbon atoms. They include the following mentioned alkyl radicals.  

Saturated hydrocarbon radicals can be linear, branched or cyclic and have up to 24 carbon atoms.

Unsaturated hydrocarbon residues can be linear, branched or cyclic and contain one or more C-C double and / or triple bonds and up to Have 24 carbon atoms.

Alkyl and the alkyl parts in alkoxy, alkylthio, mono- and dialkylamino, alkyl sulfonyl include z. B. C ₁ -C ₂₄ -, preferably C ₁ -C ₁₂ - and C ₇ -C ₂₄ -, in particular C ₁ -C ₆ - and C ₁ -C ₄ alkyl.

C ₁ -C ₂₄ alkyl includes methyl, ethyl, n- and i-propyl, n-, i-, sec- and tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, 2-ethylhexyl, n-octyl , Decyl, dodecyl, palmityl, stearyl.

C ₁ -C ₂₄ alkyl includes methyl, ethyl, n- and i-propyl, n-, i-, sec- and tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, 2-ethylhexyl, n-octyl , Decyl, dodecyl.

C ₁ -C ₆ alkyl includes methyl, ethyl, n- and i-propyl, n-, i-, sec- and tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl.

C ₁ -C ₄ alkyl includes methyl, ethyl, n- and i-propyl, n-, i-, sec- and tert-butyl.

C ₇ -C ₂₄ alkyl includes n- and i-octyl, decyl, dodecyl, palmityl, stearyl.

Cyclic hydrocarbon radicals can be mono- or polycyclic, saturated or be unsaturated, non-aromatic or aromatic and have up to 14 carbon atoms.

Aromatic cyclic hydrocarbons are preferred in the context of the invention residues, especially phenyl.  

A 5- to 8-membered saturated or partially unsaturated heterocycle with up to 3 heteroatoms from the series S, N and / or O is within the scope of the invention in general for a heterocycle which ent or one or more double bonds can hold and ver over a ring carbon atom or a ring nitrogen atom is knotting. Examples include: tetrahydrofur-2-yl, tetrahydrofur-3-yl, Pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, pyrrolin-1-yl, piperidin-1-yl, Piperidin-3-yl, 1,2-dihydropyridin-1-yl, 1,4-dihydropyridin-1-yl, piperazin-1-yl, Morpholin-1-yl, azepin-1-yl, 1,4-diazepin-1-yl. Piperidin-1-yl are preferred, Piperidin-3-yl, morpholin-1-yl.

In the context of the invention, (C ₁ -C ₇ ) -alkanoyl represents a straight-chain or branched alkyl radical having 1 to 7 carbon atoms, which carries a double-bonded oxygen atom in the 1-position and is linked via the 1-position. Examples include: formyl, acetyl, propionyl, n-butyryl, i-butyryl, pivaloyl, n-hexanoyl, n-heptanoyl.

In the context of the invention, (C ₁ -C ₆ ) alkoxycarbonyl represents a straight-chain or branched alkoxy radical having 1 to 6 carbon atoms, which is linked via a carbonyl group. A straight-chain or branched alkoxycarbonyl radical having 1 to 4 carbon atoms is preferred. Examples include: methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl and t-butoxycarbonyl.

Hydroxy protecting group in the context of the definition given above is in general NEN for a protective group from the series: trimethylsilyl, triisopropylsilyl, tert-butyl dimethylsilyl, benzyl, benzyloxycarbonyl, 2-nitrobenzyl, 4-nitrobenzyl, tert. butyl oxycarbonyl, allyloxycarbonyl, 4-methoxybenzyl, 4-methoxybenzyloxy-carbonyl, Tetrahydropyranyl, formyl, acetyl, trichloroacetyl, 2,2,2-trichloroethoxycarbonyl, Methoxyethoxymethyl, [2- (trimethylsilyl) ethoxy] methyl, benzoyl, 4-methylbenzoyl, 4-nitrobenzoyl, 4-fluorobenzoyl, 4-chlorobenzoyl or 4-methoxybenzoyl. Prefers  are acetyl, tert. Butyldimethylsilyl or tetrahydropyranyl. As part of the Erfin The tert-butyldimethylsilyl group is particularly preferred.

The present invention preferably relates to aryl-substituted oxazolidinones of the general formula (I) in the
d represents an aromatic hydrocarbon radical, preferably phenyl, which in turn can be substituted by one or more radicals R ^d , in which b represents CH ₂ and
m can have the values 0, 1 or 2,
in which a can be hydrogen, a hydrocarbon radical or a saturated or partially unsaturated heterocycle, preferably via the 2- or 4-linked pyridine or piperidine;
in which e can be amino, mono- or di- (C ₁ -C ₄ ) -alkylamino, hydroxy, (C ₁ -C ₄ ) -alkoxy which in turn can be substituted by a carbon residue, preferably phenyl,
where if d is phenyl, b is preferably ortho or para to d, and if m = 0, a is preferably ortho or para to d;
where e can stand for a group (eI)

where x or y can represent oxygen or nitrogen,
with the requirements
that when x is nitrogen R ^{e3 is} hydrogen or a hydrocarbon radical, preferably (C ₁ -C ₄ ) alkyl,
that when x and y stand for oxygen, R ^e2 and R ^e3 are omitted,
that when y is oxygen, R ^{e2 is} absent,
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₄ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together including y a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, preferably morpholin-1-yl,
R ^d can be selected from one of the groups
-COH, -NO ₂ , -CF ₃ , halogen preferably chlorine, -OR ^d1 , (C ₁ -C ₇ ) alkanoyl, alkyl, preferably (C ₁ -C ₄ ) alkyl, which in turn can be substituted, with groups selected from hydroxy,

in which
R ^d1 can represent a hydrocarbon radical, preferably phenyl or (C ₁ -C ₄ ) alkyl, which in turn can be substituted;
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon radical, (C ₁ -C ₆ ) alkoxycarbonyl or a group

wherein R ^d4 , R ^d5 can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₆ ) alkoxycarbonyl, and their salts.

The present invention particularly preferably relates to aryl-substituted oxazolini dinones of the general formula (I) in the
d represents phenyl, which in turn can be substituted by one or more radicals R ^d in which b represents CH ₂ and
m can assume the values 0 or 1,
in which a can be hydrogen, or a saturated or partially unsaturated heterocycle, preferably via the 2- or 4-linked pyridine or piperidine;
in which e can be amino, mono- or di- (C ₁ -C ₄ ) -alkylamino, hydroxy, (C ₁ -C ₄ ) -alkoxy which in turn can be substituted by a hydrocarbon radical, preferably phenyl,
in which b is preferably connected ortho or para to d, and in the event that m = 0, a is preferably connected ortho or para to d;
where e can stand for a group (eI)

where x or y can represent oxygen or nitrogen,
with the requirements
that when x is nitrogen, R ^{e3 is} hydrogen or (C ₁ -C ₄ ) alkyl;
that when x and y are oxygen, R ^e2 and R ^e3 are omitted;
that when y is oxygen, R ^{e2 is} absent;
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₄ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together including y a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, preferably morpholin-1-yl,
that R ^d can be selected from one of the groups
-COH, -NO ₂ , -CF ₃ , halogen preferably chlorine, -OR ^d1 , (C ₁ -C ₇ ) alkanoyl, alkyl, preferably (C ₁ -C ₄ ) alkyl, which in turn can be substituted with groups from selected from hydroxy,

in which
R ^d1 can represent a hydrocarbon radical, preferably phenyl or (C ₁ -C ₄ ) alkyl,
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon radical, (C ₁ -C ₆ ) alkoxycarbonyl or a group

wherein R ^d4 , R ^d5 can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₆ ) alkoxycarbonyl, and their salts.

In addition, processes for the preparation of the compounds according to the invention were found, characterized in that

• 1. [A] compounds of the general formula (AI)
  wherein
  R ^{AI, 1 represents} a hydroxy protecting group, for example a benzyl group and
  R ^{AI, 2 represents} a leaving group, for example a halogen atom, in particular a bromine atom,
  in DMF or an equivalent solvent with trialkylstan nylpyridinen for example trimethylstannylpyridines in the presence of reducing agents such as palladium (II) complexes, for example bis (triphenylphosphine) palladium (II) chloride, and a base, for example triethylamine, to give compounds of general formula (AII)
  are implemented and the hydroxy protecting group R ^{AI, 1 is} cleaved from them, so that the compounds of the general formula (AIII)
  are obtained, and in the event that R ^{AI, 1 stands} for the benzyl group, this is split off by compounds of the general formula (AII) dissolved in a polar solvent, for example an alcohol, for example ethanol, in the presence of an acid , for example hydrochloric acid and a transition metal catalyst, for example palladium, and are hydrogenated
• 2. [B] Compounds of the general formula (AIII) in polar solvents, such as DMF or an equivalent solvent, with isocyanates, for example ethyl isocyanate in the presence of a deprotonating agent, for example phosphazene base P ₁ -tBu, to give compounds with the general formula (BI)
  wherein R ^{BI, 1 represents} a hydrocarbon radical, preferably (C ₁ -C ₆ ) alkyl,

be implemented.

The processes according to the invention are illustrated by the formula scheme below.

Depending on the individual process steps, suitable solvents are suitable the usual solvents that do not change under the reaction conditions. These include non-polar solvents, polar aprotic solvents and protic solvents medium.

Examples of non-polar solvents are: aliphatic and aromatic Hydrocarbons such as benzene, toluene, xylene, hexane, cyclohexane or petroleum petroleum tions; aliphatic and aromatic halogenated hydrocarbons such as dichloromethane, Trichloromethane, carbon tetrachloride, trichloroethane, carbon tetrachloride, 1,2-dichloroethane or trichlorethylene, dichlorobenzene; Ethers such as diethyl ether, tert-butyldimethyl ether, dioxane, tetrahydrofuran (THF), glycol dimethyl ether (1,2-dimethoxyethane, "glyme" or "DME") or diethylene glycol dimethyl ether ("diglyme").

Examples of polar aprotic solvents are: ketones such as acetone or butanone, acetic acid esters such as ethyl acetate or butyl acetate [which?], amides such as dimethylformamide or hexamethylphosphoric triamide, N-methylpyrrolidone ("NMP"), nitriles such as acetonitrile, nitro compounds such as nitromethane, dimethyl sulfate oxide (DMSO), basic solvents such as pyridine, picoline, N-methylpiperidine.

Examples of protic solvents are: water or alcohols such as Methanol, ethanol, n-propanol, iso-propanol, n-butanol or tert-butanol.

The solvents can also be used in the form of mixtures. Preferred solvents are dimethylformamide (DMF) and ethanol.

All implementations are generally in normal, elevated or in ernie third pressure carried out (e.g. 0.5 to 5 bar). Generally one works at Normal pressure. Unless otherwise stated, the reactions are at room temperature temperature carried out.  

The compounds described above can be used to prepare medicinal products be used for the treatment of diseases in humans and animals.

The compounds described above are inhibitors of TNFα Biosyn thesis. They can be used in the prevention and treatment of Cardiovascular diseases, such as B. atherosclerosis and arthritis, rheumatoid arthritis, osteoporosis, Crohn's disease, chronic inflammatory Lung diseases such as Adult Respiratory Distress Syndrome (ARDS), graft Rejection, reperfusion tissue damage after stroke, heart attack or peripheral vascular occlusions, chronic inflammatory fibrotic Organ changes such as liver fibrosis, or the generalized Autoimmune disease systemic lupus erythematosus or other forms of Lupus erythematosus and dermal inflammatory diseases such as psoriasis.

The inhibition of TNFα biosynthesis by the verb described above dungs can be demonstrated in biological tests in vitro and in vivo.

The active ingredient can be administered orally or parenterally to achieve a systemic effect, can be applied locally for an external effect.

Application forms on the are particularly suitable for parenteral administration Mucous membranes (buccal / lingual / sublingual, rectal, nasal, pulmonal, conjunctival or intravaginal) or inside the body. This can be done by bypassing absorption happen (intracardially, intraarterially, intravenously, intraspinally or intralumbally) or with absorption activated (intracutaneous, subcutaneous, percutaneous, intra muscular or intraperitoneal).

The active ingredients can be administered alone or in the form of administration forms be enough.  

For oral application are suitable as forms of application u. a. normal and enteric-coated tablets, capsules, dragees, pills, granules, pellets, powders, solid and liquid aerosols, syrups, emulsions, suspensions and solutions.

For parenteral administration, injection and administration forms are suitable Infusion solutions.

Suppositories are suitable for local application with systemic distribution and aerosols, if no systemic distribution is desired, vaginal are suitable capsules, nose, ear, eye drops, aqueous suspensions (lotions, shakes mixtures), lipophilic suspensions, ointments, creams, milk, pastes, powder or Sprays.

In the application forms, the active ingredient can be used in a concentration of 0-100% by weight are present; preferably the concentration of the active ingredient 0.5-90 wt%, i.e. H. in amounts that are sufficient Achieve dosage range.

The active substances can be added to the applications listed in a manner known per se forms are transferred. This is done using inert non-toxic, pharmaceutically suitable excipients, e.g. B. carriers, solvents, vehicles, Emulsifiers and / or dispersants.

Examples of auxiliaries include: water, solid carriers such as natural Liche or synthetic stone powder (e.g. talc or silicates), sugar (e.g. Milk sugar), non-toxic organic solvents such as paraffins, vegetable oils (e.g. sesame oil), alcohols (e.g. ethanol, glycerin), glycols (e.g. polyethylene glycol), Emulsifiers, dispersants (e.g. polyvinylpyrrolidone) and lubricants (e.g. Magnesium sulfate).  

In the case of oral administration, tablets can of course also contain additives like sodium citrate along with additives like starch, gelatin and the like Chen included. Aqueous preparations for oral administration can continue with flavor enhancers or colorants.

In general, it has proven to be advantageous for parenteral administration Amounts from about 0.001 to 25 mg / kg, preferably about 0.001 to 10 mg / kg Give body weight for effective results. With oral Application, the amount is about 0.001 to 50 mg / kg, preferably about 0.01 to 20 mg / kg body weight.

Nevertheless, it may be necessary from the amounts mentioned deviate, depending on body weight, application route, individual behavior towards the active ingredient, type of preparation and time point or interval at which the application takes place.

1. Detection of TNFα biosynthesis inhibition in human blood monocytes

Peripheral blood monocytes are made from 150 ml of blood from healthy volunteers as follows isolated:

The blood is in Vacutainer tubes (Becton-Dickinson GmbH, Heidelberg, Order No. 362753) centrifuged for 20 minutes at 1500 g at room temperature. The Monocytes are removed from a zone above the cell according to the manufacturer's instructions Aspirated separating gel and in a 50 ml Falcon centrifuge tube in 50 ml phos phat-buffered saline suspended and centrifuged again at 300 g for 20 minutes.

The cell pellet is dissolved in 10 ml of Versene solution (Life-Technologies GmbH, Karls rest, order no. 15040-033) and centrifuged again at 300 g for 5 minutes. The cells are then placed in culture medium RPMI 1640 (from Life-Technolo gies GmbH, Karlsruhe, order no. 21875-059) added. For the culture medium  1% (vol / vol) of the following solutions are used as additives: 200 mM L-glutamine; 5000 U / ml penicillin G and 5 mg / ml steptomycin sulfate; 1.0 M HEPES buffer solution; 100 mM sodium pyruvate; Solution; nonessential Amino acids (Life-Technologies GmbH, Karlsruhe, order no.11140-035) Furthermore, 10% (vol / vol) fetal calf serum is added to the culture medium puts.

The monocytes are sown in 100 μl in this culture medium with a cell density of 2.5 × 10 ⁵ cells / well in a 96-well microtiter plate (Falcon / Becton Dickinson, Lincoln Park, NJ 07035, Microtest III, Tissue Culture Plate) , After 1 to 2 hours, the monocytes adhere to the surface. The medium is suctioned off and the monocytes are washed with 100 ul culture medium. Subsequently, 150 μl of culture medium, the test substance, are dissolved in 50 μl of culture medium and 50 μl of lipopolysaccharide (LPS) solution (0.5 μg / ml of LPS in culture medium: Escherichia coli Serotype 0111: B4 from Fa. Sigma, St. Louis, MO 63178, USA, Order No. L-3012) was added and incubated for 18 hours in a cell culture incubator at 37 ° C. and 5% CO2 atmosphere.

The amount of TNFα formed is in 100 µl supernatant of the test batches with a commercially available enzyme-linked immunosorbent test (ELISA), e.g. B. from from R Systems GmbH, 65205 Wiesbaden, order no. DTA50, according to An would be determined quantitatively by the manufacturer. The TNFα synthesis inhibition can in this way depending on the concentration of the test substance used voices. The concentration of the test substance, which is a half-maximum TNFα Synthesis inhibition (EC50) is caused by the corresponding dose-response curve determined.

Table 1

TNF ELISA: Effective data

2. Evidence of inhibition of TNF release in vivo

Female Balb / c or OF- were used to determine the TNF release in vivo. 1 mice (Iffa creed) used. These were checked 4 hours before the start of the experiment sober.

The substances to be tested were administered by os, intraperitoneally or intravenously enough. The application volume was peroral and intraperitoneal 10 ml / kg, whereas only 5 ml / kg were administered with intravenous administration.

The TNF release in the mice was characterized by the application of LPS S. minnesota (4 mg / kg) induced the intraperitoneally in a volume of 25 ml / kg was administered in 0.9% NaCl. LPS administration was routinely 30 minutes later the oral administration of substance. For the analysis of kinetic questions or after intravenous administration, LPS was injected 15 to 120 minutes after substance administration.

In all studies, the mice were administered retro-orbi 1.5 hours after LPS administration tale puncture blood was taken and sent for further analysis. That comes from My blood was centrifuged at 1400 rpm for 10 minutes to collect serum. Using a TNF ELISA from R, the TNF content determined.  

3. Evidence of the anti-atherosclerotic effects of the above described compounds and TNF synthesis inhibitors in the model the ApoE knockout mouse

To determine the anti-atherosclerotic activity of the compounds, the Model of the ApoE deficient mouse used. This model is mice that have a disturbed gene due to a gene defect in apolipoprotein E. Have lipid metabolism and within 3 months in the heart valve area develop a form of atherosclerosis, that of advanced atherosclerosis of humans is very similar. This model has become one in recent years Standard model of atherosclerosis research developed.

To quantify atherosclerosis, the area of interest is the heart flap removed and fixed in paraffin. Then in histological Cut the cross-sectional area of the atherosclerotic deposits with the help of a computer-based morphometry system  

Abbreviations

abs .: absolutely
Ac: acetyl
aq .: watery
Boc: tert-butoxycarbonyl
Bu: butyl
CDI: N, N'-carbonyldiimidazole
TLC: thin layer chromatography
DCC: N, N'-dicyclohexylcarbodiimide
DEAD: diethyl azodicarboxylate
dist .: distilled
DIBAH: diisobutyl aluminum hydride
DMAP: 4-N, N-dimethylaminopyridine
DME: 1,2-dimethoxyethane
DMF: N, N-dimethylformamide
DMPU: N, N'-dimethylpropyleneurea
DMSO: dimethyl sulfoxide
DPPA: diphenylphosphoryl azide
EDC: N '- (3-dimethylaminopropyl) -N-ethylcarbodiimide × HCl
eq: equivalent (s)
Et: ethyl
fl .: liquid
Mp .: melting point
Fr .: fraction
GC: gas chromatography
sat .: saturated
HMPT: hexamethylphosphoric triamide
HOBt: 1-hydroxy-1H-benzotriazole × H2O
HOSu: N-hydroxysuccinimide
HPLC: high pressure, high performance liquid chromatography
conc .: concentrated
Kp .: boiling point
krist .: crystalline / crystallized
LAH: lithium aluminum hydride
LDA: Lithium-N, N-diisopropylamide
LiHMDS: Lithium-N, N-bistrimethylsilylamide
Lit .: literature (place)
Solution: solution
MCPBA: meta-chloroperbenzoic acid
Me: methyl
MEK: methyl ethyl ketone
MPLC: medium pressure liquid chromatography
MS: mass spectroscopy
MTBE: methyl tert-butyl ether
Nd .: precipitation
NBS: N-bromine succinimide
NMM: N-methylmorpholine
NMR: nuclear magnetic resonance spectroscopy
p. A .: pro analysi
Ph: phenyl
pr: propyl
RF: reflux
RF: retention index (for DC)
RT: room temperature
subl .: sublimed
TBAF: tetrabutylammonium fluoride
TBDMS: tert-butyldimethylsilyl
TEA: triethylamine
techn .: technical
TFA: trifluoroacetic acid
TFAA: trifluoroacetic anhydride
THF: tetrahydrofuran
titr .: titrated
TMS: trimethylsilyl
TPP: triphenylphosphine
TPPO: triphenylphosphine oxide
dil .: diluted
Vol .: volume
watery: watery
Z: benzyloxycarbonyl
Decomposition: decomposition

The eluents used for thin layer chromatography are:
A = dichloromethane / methanol = 96/4
B = dichloromethane / methanol = 98/2
C = toluene / ethanol = 10/1
D = dichloromethane / methanol = 9/1

starting compounds

Example I

5 - [(benzyloxy) methyl] -3- [4- (4-pyridinyl) phenyl] -1,3-oxazolidin-2-one

2.0 g (5.52 mmol) 5 - [(benzyloxy) methyl] -3- (4-bromophenyl) -1,3-oxazolidin-2-one (synthesis of the starting material was carried out according to: 5. Raddatz, 5. Bartel , W. Guarnieri, U. Rosentreter, M. Ruppelt, H. Wild, R. Endermann, H.-P. Kroll, K. Henninger, WO 99 40094 A1) are treated under argon in 30 mL anhydrous DMF with 1.6 g (6.63 mmol) freshly prepared 4-trimethylstannyl-pyridine (synthesis: JE Phillips, RH Herber, J. Organomet. Chem. 268, 39 (1984).), 0.28 g (0.39 mmol) bis- ( triphenylphosphine) palladium (II) chloride and 1.37 g (13.5 mmol) of triethylamine are added and the mixture is stirred at 60 ° C. for 48 hours. The DMF is then evaporated in a high vacuum at a maximum of 40 ° C. and the residue is chromatographically separated into its components (silica gel 60, Merck, dichloromethane: methanol = 99: 1).
Yield: 1.9 g (95%)
R _f = 0.19 (B)
MS (FAB): 361 [M + H] +

The compounds in the following table are prepared analogously to the starting compound IV.

Example V

5- (hydroxymethyl) -3- [4- (4-pyridinyl) phenyl] -1,3-oxazolidin-2-one

0.37 g (1.02 mmol) of compound IV are dissolved in 100 ml of ethanol, adjusted to pH = 2-3 with 6N hydrochloric acid and hydrogenated in the presence of 0.2 g of palladium / black as catalyst for 18 hours at room temperature. The mixture is filtered off through Celite and the solvent is distilled off in vacuo. The residue is taken up in dichloromethane and the organic phase is washed with saturated sodium carbonate solution, dried over sodium sulfate and concentrated.
Yield: 0.27 g (97%)
R _f = 0.17 (A)

The compounds in the following table are prepared analogously to the starting compound VIII.

Preparation Examples

example 1

{2-Oxo-3- [4- (4-pyridinylmethyl) phenyl] -1,3-oxazolidin-5-yl} methyl ethyl carbamate

0.13 g (0.45 mmol) of the compound of the example are dissolved in 3 ml. DMF, with 0.04 g (0.56 mmol) of ethyl isocyanate and a drop of N '''- tert-butyl-N, N, N' , N ', N ", N" -hexamethylphosphorimidsäuretriamid (phosphazene base P1-tBu) and stirred overnight at 23 ° C. The reaction mixture is poured into water and the precipitate is filtered off with suction and dried.
Yield: 0.11 g (69%)
Mp = 140 ° C
Rf = 0.12 (A)

The compounds in the following table are prepared analogously to the procedure in Example 1.

Example 5

{(5R) -2-Oxo-3- [3- (4-pyridinyl) phenyl] -1,3-oxazolidin-5-yl} methyl methyl carbamate hydrochloride

0.02 g (0.061 mmol) {(5R) -2-oxo-3- [3- (4-pyridinyl) phenyl] -1,3-oxazolidin-5-yl} methyl methyl carbamate are dissolved in 2 ml dichloromethane / methanol ( 1: 1) dissolved and mixed with 0.5 ml of 1 N hydrochloric acid. The solvent is then distilled off and the residue is dried in vacuo.
Yield: quant.
Mp .: <250 ° C

Claims (7)

1. Aryl-substituted oxazolidinones of the general formula (I)
in the
d represents a cyclic hydrocarbon radical, for example an aryl radical, which in turn can be substituted by one or more radicals R ^d ,
where b is CH ₂ and
m can assume the values 0 to 4,
where a can be hydrogen, a hydrocarbon radical or a saturated or partially unsaturated or aromatic heterocycle, having 1 to 3 heteroatoms selected from S, N, O;
in which e can stand for amino, mono- or di-substituted amino, hydroxy, (C ₁ -C ₆ ) alkoxy which in turn can be substituted with a hydrocarbon radical, for example an aryl radical, or a hydroxy protective group;
where e can stand for a group (eI)
wherein
x or y can represent oxygen or nitrogen,
with the requirements
that when x is nitrogen R ^{e3 is} hydrogen or a hydrocarbon radical, preferably (C ₁ -C ₆ ) alkyl,
that when x and y stand for oxygen, R ^e2 and R ^e3 are omitted,
that when y is oxygen, R ^{e2 is} absent,
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₆ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together with y including a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, which in turn can be further substituted;
in the round can be selected from one of the groups
-COH, -NO ₂ , -CX ₃ , where X is halogen, -OR ^d1 , (C ₁ -C ₈ ) alkanoyl, alkyl, preferably (C ₁ -C ₆ ) alkyl, which in turn can be substituted with Groups selected from hydroxy,
in which
R ^d1 can represent a hydrocarbon radical, for example an aryl radical or (C ₁ -C ₆ ) alkyl, which in turn can be substituted;
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon radical, (C ₁ -C ₈ ) alkoxycarbonyl or a group
where R ^d4 , R ^d5 can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₈ ) alkoxycarbonyl,
and their salts. 2. Aryl-substituted oxazolidinones of the general formula (I) in the
d represents an aromatic hydrocarbon radical, preferably phenyl, which in turn can be substituted by one or more radicals R ^d ,
where b is CH ₂ and
m can have the values 0, 1 or 2,
where a can stand for hydrogen, a hydrocarbon residue or a saturated or partially unsaturated heterocycle, preferably via the 2- or 4-linked pyridine or piperidine;
in which e can be amino, mono- or di- (C ₁ -C ₄ ) -alkylamino, hydroxy, (C ₁ -C ₄ ) -alkoxy which in turn can be substituted by a hydrocarbon radical, preferably phenyl,
where if d is phenyl, b is preferably ortho or para to d, and if m = 0, a is preferably ortho or para to d;
where e can stand for a group (eI)
where x or y can represent oxygen or nitrogen,
with the requirements
that when x is nitrogen R ^{e3 is} hydrogen or a hydrocarbon radical, preferably (C ₁ -C ₄ ) alkyl,
that when x and y stand for oxygen, R ^e2 and R ^e3 are omitted,
that when y is oxygen, R ^{e2 is} absent,
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₄ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together including y a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, preferably morpholin-1-yl,
R ^d can be selected from one of the groups
-COH, -NO ₂ , -CF ₃ , halogen preferably chlorine, -OR ^d1 , (C ₁ -C ₇ ) alkanoyl, alkyl, preferably (C ₁ -C ₄ ) alkyl, which in turn can be substituted with groups selected from hydroxy,
in which
R ^d1 can represent a hydrocarbon test, preferably phenyl or (C ₁ -C ₄ ) alkyl, which in turn can be substituted;
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon radical, (C ₁ -C ₆ ) alkoxycarbonyl or a group
where R ^d4 , R ^d5
can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₆ ) alkoxycarbonyl,
and their salts. 3. Aryl-substituted oxazolidinones of the general formula (I) in the
d represents phenyl, which in turn can be substituted by one or more radicals R ^d ,
n the b stands for CH ₂ and
m can assume the values 0 or 1,
in which a can be hydrogen, or a saturated or partially unsaturated heterocycle, preferably via the 2- or 4-linked pyridine or piperidine;
where e can be amino, mono- or di- (C ₁ -C ₄ ) -alkylamino, hydroxy, (C ₁ -C ₄ ) -alkoxy which in turn can be substituted with a hydrocarbon radical, preferably phenyl,
in which b is preferably connected ortho or para to d, and if m = 0, a is preferably connected ortho or para to d;
where e can stand for a group (eI)
where x or y can represent oxygen or nitrogen,
with the requirements
that when x is nitrogen, R ^{e3 is} hydrogen or (C ₁ -C ₄ ) alkyl;
that when x and y are oxygen, R ^e2 and R ^e3 are omitted;
that when y is oxygen, R ^{e2 is} absent;
that R ^e1 , R ^{e2 can be} hydrogen or (C ₁ -C ₄ ) alkyl, or that when y is nitrogen R ^e1 and R ^e2 together including y a 5 to 8-membered, saturated or partially unsaturated heterocycle can form with up to 3 heteroatoms from the series S, N, O, preferably morpholin-1-yl,
that R ^d can be selected from one of the groups
-COH, -NO ₂ , -CF ₃ , halogen preferably chlorine, -OR ^d1 , (C ₁ -C ₇ ) alkanoyl, alkyl, preferably (C ₁ -C ₄ ) alkyl, which in turn can be substituted with groups selected from hydroxy,
in which
R ^d1 can represent a hydrocarbon radical, preferably phenyl or (C ₁ -C ₄ ) alkyl,
R ^d2 , R ^d3 can represent hydrogen, a hydrocarbon test, (C ₁ -C ₆ ) alkoxycarbonyl or a group
where R ^d4 , R ^d5 can represent hydrogen, a hydrocarbon radical or (C ₁ -C ₆ ) alkoxycarbonyl,
and their salts. 4. Compounds according to any one of claims 1 to 3 for use in the Fighting disease. 5. Use of compounds according to one of claims 1 to 3 for Her provision of medicines. 6. Use of compounds according to one of claims 1 to 3 for Her Provision of medicines that have a preventive, soothing or healing effect disease patterns mediated by TNFα such as arthritis, rheumatoid Arthritis, osteoporosis, Crohn's disease, chronic inflammatory lungs diseases such as Adult Respiratory Distress Syndrome (ARDS), graft  Rejection, reperfusion tissue damage after stroke, heart attack or peripheral vascular occlusions, chronic inflammatory fibrotic organ changes like liver fibrosis, or the generalized Antoimmun systemic lupus erythematosus or other forms of Lupus erythematosus and dermal inflammatory diseases such as psoriasis. 7. Medicament containing a compound according to any one of claims 1 to 3 and a pharmaceutically acceptable auxiliary.