WO2003068225A1

WO2003068225A1 - Microglia inhibitors for interrupting immune reactions induced by interleukin 12 and ifn$g(g)

Info

Publication number: WO2003068225A1
Application number: PCT/EP2003/000467
Authority: WO
Inventors: Thorsten Blume; Wolf-Dietrich DÖCKE; Wolfgang Halfbrodt; Joachim Kuhnke; Ursula MÖNNING; Bernd Elger; Herbert Schneider
Original assignee: Schering Aktiengesellschaft
Priority date: 2002-02-15
Filing date: 2003-01-17
Publication date: 2003-08-21
Also published as: JP2005522447A; RS71304A; UY27662A1; ZA200407382B; MXPA04007943A; PE20030906A1; NO20043840L; AU2003245523A1; RU2004127677A; AR039556A1; UA81111C2; CA2475770A1; ECSP045297A; KR20040084909A; PL371327A1; TW200306823A; CN1756546A; BR0307706A; EP1474138A1; DE10207843A1

Abstract

The invention relates to the use of microglia inhibitors for producing medicaments that inhibit the monocyte, macrophage and T cell-induced immune reactions, and to their use for treating T cell-induced immunological diseases and inflammatory reactions that are not T-cell induced.

Description

Microglial inhibitors for interruption of interleukin 12 and IFNγ mediated immune responses

The invention relates to the use of a microglial inhibitor for the manufacture of a medicament which inhibits interleukin 12 (IL 12) and interferon-γ mediated immune responses, and its use for the treatment of T cell mediated immunological disorders and non-T cell mediated inflammatory responses ,

The immune system includes a variety of cells and tissue complexes that communicate primarily via soluble factors. It is known that many immunological diseases are characterized by an imbalance of soluble immune factors, such as. Rev. Immunol., 7: 145-173 (1989, Street and Mosman, FASEB J. 5: 171-177 (1991); Lucey et al., Clin. Microbiol. Rev. 4: 532-562 (1996); Powrie and Coffman, Trends Pharmacol., 14: 164-168 (1993); Singh et al., Immunolog Res., 20: 164-168 (1999)) For example, there are a number of indications of a role of interferon gamma and interleukin 12 in the pathogenesis of autoimmune diseases, in particular diseases characterized by a T cell-mediated inflammatory reaction, such as multiple sclerosis, diabetes, Inflammatory Bowel Diseases The cytokine interleukin 12 (IL 12) is produced by phagocytic cells, such as macrophages / monocytes, dendrites, B cells, and other antigen presenting cells (APCs), and affects both function of natural killer cells (NK cells) as well as of T lymphocytes IL 12 can stimulate the production of interferon gamma (IFNγ) in both cell types. T lymphocytes can be roughly classified into two categories characterized by the expression of certain surface antigens (CD4 and CD8): CD4-positive T cells (helper T cells) and CD8-positive T cells (cytotoxic T cells) T-cells). The CD4 cells can in turn be divided into T helper cells 1 (Th1) and T helper cells 2 (Th2). Especially the Th1-mediated immune responses are more numerous with the pathogenesis Immune disorders, especially autoimmune diseases, such as: type I insulin-dependent diabetes mellitus (IDDM), multiple sclerosis, allergic contact dermatitis, psoriasis, rheumatoid arthritis, chronic inflammatory bowel disease (Crohn's disease, ulcerative colitis) ), Lupus diseases and other collagenoses as well as acute graft-versus-host disease ("host-versus-graft") allografts. Interleukin 12 is known to play a critical role in regulation Interleukin 12 induces production of primarily IL-2, IFNγ, TNFα and TNFβ in these cells (Mosmann and Sad, Immunol., Today 17: 138-146 (1996); Gately et al., Annu. Rev Immunol., 16: 495-521 (1998)). Specifically, IFNγ is a potent mediator of IL-12 activity. Overproduction of interferon gamma may be exemplified by MHC II (Major Histocompatibility Comp lex) -associated autoimmune diseases (. In addition, there is also sufficient evidence regarding a pathological role of interferon gamma in allergic diseases as well as sarcoidosis and psoriasis (Billiau A., Adv. Immunol., 62: 61-130 (1996); Basham et al., J. Immunol Hu et al., Immunology, 98: 379-385 (1999); Seery JP, Arthritis Res., 2: 437-440 (2000)). In addition, IL-12 and IL ^' -12 / IL-18-induced IFNγ from NK cells are essential in the pathomechanism of non-T cell-mediated inflammatory responses (eg, "Toxic shock syndrome", endotoxemia, sepsis and septic shock, ARDS, "first dose response "in antibody therapy eg OKT3 administration in allotransplantation) (Kum et al., Infect Immun. 69: 7544-7549 (2001); Arad et al., J Leukoc. Biol. 69: 921-927 (2001); Huitgren et al., Arthritis Res. 3: 41-47 (2001), Arndt et al., Am J. Respir, Cell Mol. Biol. 22: 708-713 (2000); Grohmann et al., J. Immunol 164: 4197-4203 (2000); Muraille et al., Int. Immunol., V: 1403-1410 (1999)). IL-12 also plays a role in inflammation with currently unclear pathomechanisms (eg, eclampsia) (Hayakawa et al., J. Reprod Immunol 47: 121-138 (2000); Daniel et al., Am J. Reprod., Immunol 39: 376-380 (1998)). In addition to interleukin 12 and IFNγ, other cytokines will also play a role in

Pathogenesis of immune diseases and systemic

Attributed to inflammatory reactions, such as TNFα. TNFα plays a significant pathological role in infectious diseases (such as sepsis, "toxic shock syndrome" (Tracey et al., Nature 330: 662-664 (1987);

Basger et al., Circ. Shock, 27: 51-61 (1989); Hinshaw et al., Circ. Shock, 30:

279-292 (1990); Libra A., Lancet, 351: 603 (1998); Cohen et al., Lancet,

351: 1731 (1998)), but also numerous other immune-mediated

Diseases.

For the treatment of IL 12-mediated diseases and for the reduction of the acute symptoms of these diseases corticosteroids are often used, the side effects, especially in long-term treatment often lead to discontinuation of treatment.

Activation of the microglia is a central step in the inflammatory process of almost all degenerative diseases of the central nervous system. The microglia may remain in the activated state for a prolonged period of time by causing various inflammatory factors, e.g. B. reactive oxygen / nitrogen intermediates,

Proteases, cytokines, complement factors and neurotoxins, produce and secrete. These in turn cause neuronal dysfunction and degeneration. The activation of microglia can be done by various stimuli, such as. 569: 141-145 (1992)), phonon protein, cytokines, or by cell fragments (Combs, CK et al., J. Neurosci 19: 928-939 (1999); Wood, P.L., Neuroinflammation: Mechanisms and Management, Humana Press (1998).

Compounds which inhibit the activation of the microglia after stimulation with the Aβ peptide are described in WO 01/51473, DE reference 101 34 775.8 and DE reference 101 35 050.3. It is also known that benzimidazoles which inhibit the activation of microglia are useful in the treatment of neuroinflammatory diseases such as AIDS dementia, amyotrophs Lateral Sclerosis, Creutzfeldt-Jakob Disease, Down's Syndrome, Diffuse Lewy Body Disease, Huntington's Disease, Leukencephalopathy, Multiple Sclerosis, Parkinson's Disease, Pick's Disease, Alzheimer's Disease, Stroke, Temporary Lobe Epilepsy, and Tumors.

The invention is based on the problem of providing a means suitable for the treatment of immunological diseases wherein the immunological diseases are triggered by increased production of cytokines, e.g. IL 12, IFN and TNFα without other factors in the

Immune system act, whereby side effects are reduced or prevented.

It has now been found, surprisingly, that microglial inhibitors inhibit the production of IL 12 and IFNγ and induce interleukin-10 production (IL 10) and are therefore used for the preparation of a medicament for the treatment of a monocyte, macrophage or T cell - Mediated, specifically Th1-cell-mediated, immunological disease and non-T-cell-mediated pathophysiological inflammatory reactions.

As explained above, after expression of their surface antigens, T lymphocytes can be subdivided into CD4-positive T cells (helper T cells) and CD8-positive T cells (cytotoxic T cells) and the CD4 helper cells T-cells (Th cells) turn into T-helper cells 1 (Th1) and T-helper cells 2 (Th2), which are u. a. differ in their ability to prevent tolerance. The cytokines IL 12 and IFNγ play an important role in the induction and maintenance of THI cell differentiation. Differentiated Th1 cells secrete IFNγ, interleukin 2 and TNFα / β. These cytokines in turn activate macrophages and cytotoxic CD8 positive T cells.

In particular, the invention relates to the use of microglial inhibitors for the preparation of a medicament for interrupting IL 12 or IFNγ production in cells of monocytic origin or T cells and NK cells. On Their ability to disrupt the production of IL 12 and TNFα in monocytes / macrophages / dendrites and IFNγ production in T cells and NK cells and to increase the induction of IL-10 production are numerous inhibitors of microglia Disorders which are triggered by the increased production of cytokines, such as TNFα.β, IFNγ, IL-2 and IL12, such as inflammatory diseases, which are not based on neuroinflammation, autoimmune diseases, allergic and infectious diseases, toxin-induced inflammation , pharmacologically triggered inflammatory reactions as well as pathophysiologically relevant inflammatory reactions of currently unclear origin.

Examples of inflammatory and autoimmune diseases are: chronic inflammatory bowel disease (inflammatory bowel disease, Crohn's disease, ulcerative colitis), arthritis, allergic contact dermatitis, psoriasis, pemphigus, asthma, multiple sclerosis, diabetes, type I insulin-dependent diabetes mellitus, Rheumatoid arthritis, lupus and other collagenosis, Graves' disease, Hashimoto's disease, graft versus host disease and graft rejection.

Examples of allergic, infectious and toxin-induced and ischemia-induced diseases are: Sarcoidosis, Asthma, Hypersensitive Pneumonitis, Sepsis, Septic shock, Endotoxin shock, Toxic shock syndrome, Toxic liver failure, ARDS (Acute Respiratory Syndrome), Eclampsia, Cachexia, Acute Viral infections (eg mononucleosis, fulminant hepatitis), organ damage after reperfusion.

An example of a pharmacologically triggered inflammation with pathophysiological relevance is the "first dose response" after administration of anti-T cell antibodies such as OKT3.

An example of systemic inflammatory reactions of currently unclear origin is eclampsia. Suitable microglial inhibitors according to the invention are compounds which, upon stimulation with the Aβ peptide, achieve an inhibition of the microglial activity of at least 20% and an inhibition of the cytokine activity of at least 30%. The biological properties of the microglial inhibitors can be demonstrated by the methods known to the person skilled in the art, for example with the aid of the investigation methods described below and in WO 01/51473.

Particularly suitable microglial inhibitors with the properties described above are benzimidazoles of the formula I, their tautomeric and isomeric forms and salts.

In it are:

R ^{1 is} an aryl group or a five- or six-membered one

Heteroaryl group having one or two heteroatoms selected from the group comprising N, S and O, wherein the aryl or

Heteroaryl group having up to three radicals independently of one another, selected from the group comprising:

F, Cl, Br,

C (NH) NH ₂ , C (NH) NHR ⁴ , C (NH) N /? V, C (NR ⁴ ) NH ₂ , C (N /?) NHf? ^{4 '} ,

C (Nfl *) NflV _f

X-OH, X-OR ⁴ , X-OCOR ⁴ , X-OCONHR ⁴ ,

X-COR ⁴ , XC (NOH) /? ⁴ ,

X-CN, X-COOH, X-COOR ⁴ , X-CONH ₂ , X-CON /? V, X-COHHR ⁴ , X-CONHOH,

X-SR ⁴ , X-SOR ⁴ , X-SO ₂ R ⁴ , SO ₂ NH ₂ , S0 ₂ NHR ⁴ , S0 ₂ NR ⁴ R ^{4 '} ,

N0 ₂ , X-NH ₂ , XN R ⁴ , X-NRV ^' , X-NHSO ^, X-R ⁴ S0 ₂ R ^4' , X-NHCOK ⁴ , X-HCOOR ⁴ , X-NHCONHR ⁴ and

R ⁴ , wherein X is a bond, CH ₂ , (CH ₂ ) ₂ or CH (CH ₃ ) ₂ , wherein furthermore the radicals R ⁴ and R ⁴ are selected independently of one another according to the meanings given below and wherein two substituents on R ¹ if they are ortho on each other, can each be linked together so that they together form a Methandiylbisoxy-, ethane-1, 2-diylbisoxy, propane-1, 3-diyl or butane-1, 4-diyl group, or

a radical selected from the group comprising Ci-βalkyl, (C _0-3 alkanediyl-C _3-7 cycloalkyl) and C _3-6 alkenyl, in which one H atom is substituted by a heterocyclic radical selected from the group compound comprising piperazine, morpholine, piperidine and pyrrolidine, which may be exchanged to form a bond to a first N-atom of the heterocyclic radical, wherein said alkyl, cycloalkyl, alkenyl and heterocyclic radicals are substituted with up to two radicals selected from the group comprising C _0-2 alkanediyl-OH, Co ₂ alkanediyl-O? ⁷ ,

C _{_0-2} alkanediyl-NH _2l C _0-2 alkanediyl NHA? ^7, C _{_0-2} alkanediyl-N? ⁷ R ^{7 '} , Co ₂ -alkanediyl-NHCOR ⁷ , C _0-2 -alkanediyl-NR ⁷ COR ⁷ ', C _0-2 -alkanediyl-NHSO ₂ f? ⁷ , Co _-2 alkanediyl NR ⁷ S0 ₂ R ^{7 '} , C _0-2 alkanediyl CO ₂ H, C _0-2 alkanediyl C0 ₂ ? ^7, C ₀ - ₂ alkanediyl-CONH _2, Co ₂ alkanediyl-C _0-2 alkanediyl CONHR ⁷

CONR ⁷ ? ^{7 '} , phenyl and a five- or six-membered heteroaryl radical, wherein the heteroaryl radical contains one or two heteroatoms selected from the group comprising N, S and O, wherein furthermore the phenyl and the heteroaryl radical having up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C2F5 and S0 ₂ NH ₂ , and / or can also carry a fused Methandiylbisoxy- or ethane-1, 2-diylbisoxygruppe, wherein the Piperazinrest on a second nitrogen atom with R ⁷ , COR ⁷ or S0 ₂ R ⁷ can be substituted, wherein R ⁷ and R ⁷ 'can be selected independently of one another according to the meanings given below,

-ZR ² 'is an aryl group or a five- or six-membered one

Heteroaryl group having one or two heteroatoms selected from the group comprising N, S and O, a benzothienyl group or an indolyl group, wherein said aryl or heteroaryl group may be substituted with up to three independently selected from the group consisting of

F, Cl, Br,

C (NH) NH _2, C (NH) # *, C (NH) NRV ', C (N / ^r) NH _2, C ^(NT) NH? '

CfN / NRNRV, X-OH, X-OR ⁴ , X-OCOR ⁴ , X-OCONH ⁴ ,

X-COR ⁴ , XC (NOH)? ⁴ ,

X-CN, X-COOH, X-COOR ⁴ , X-CONH _2I X-CONR ⁴ R ^{4 '} , X-CONHR ⁴ ,

X-CONHOH,

X-SR ^4, SOR ^X-4, X-SO 2 R ^4, SO2NH2, S0 ₂ NH ^R, S0 ₂ NR ^{4 4 ',}

N0 _2, X-NH _2l X HR ^4, X-NR ⁴ R ^{4 ',} X HSO2R ^4, X-UR ⁴ S0 ₂ R ^4',

X-NHCOR, X-HCOOR ⁴ , X-NHCONHR ⁴ and

R ⁴ , wherein a bond, CH ₂ , (CH ₂ ) ₂ , or CH (CH ₃ ) ₂ , further wherein the radicals R ⁴ and R ⁴ 'are independently selected according to the meanings given below and where two radicals of R ¹ , if they are ortho on each other, can be linked to each other so that they together form a methanediylbisoxy, ethane-1,2-diylbisoxy, propane-1,3-diyl or butane-1, 4 form diyl group,

Z is NH, NR ² ", O, S, SO or SO ₂ , where R has the meaning given below,

R ² and R ² are each independently a radical selected from the group comprising:

C _1-4 perfluoroalkyl, Cι _-6 alkyl, (C _{_0-3} alkanediyl-C _3-7 cycloalkyl), (Co-3 -alkanediyl-aryl) and (C _0- alkanediyl 3-heteroaryl), wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, and wherein the aryl and heteroaryl groups each contain up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H _5> N0 ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , may be substituted and / or also can carry a fused methanediylbisoxy or ethane-1,2-diylbisoxy group, and furthermore a ring member in a five-membered cycloalkyl ring can be Ring-N or Ring-0 and one or two ring members in a six- or seven-membered cycloalkyl ring Ring-N- and / or or ring may be O-atoms, where the ring N atoms optionally substituted by Cι _-3 alkyl or Cι _-3- alkanoyl may be substituted, or R ² and R ² screens together with Z a five- to form a N-heterocyclic ring when Z is an N-atom, wherein Z has the meaning given above, wherein further the heterocyclic ring may contain another N, O or S atom and may optionally be substituted by a radical selected from of the

Group comprising Cι _-4 alkyl, (C ₀ -3-alkanediyl-C _1-3 alkoxy), Cι- - alkanoyl, C _1-4 alkoxycarbonyl, aminocarbonyl and aryl, R ^{3 are} independently one or two radicals selected from

Group comprising:

Hydrogen,

F, Cl, Br, OH, OR ⁴ , OCOR ⁴ , OCONHR ⁴ ,

COR ⁴ ,

CN, COOH, COOK ⁴ CONH ₂ , CONHR ⁴ , CONR ⁴ /? ^{4 '} , CONHOH,

CONHO /? ⁴ ,

SR ⁴ , SOR ⁴ , SO ₂ /? ⁴ , SO ₂ NH ₂ , SO ₂ NHR ⁴ , SO ₂ NR ⁴ /? ^{4 '} , NO ₂ , NH ₂ , NHR ⁴ , NR ⁴ /? ^{4 '} ,

NHSO2K ⁴ , N T'SOz? ^{4 ',} NHS0 ₂ K ^6, NR ⁴ S0 ₂ R ^6,

NHCOR ⁴ , NHCOOR ⁴ , NHCONHR ⁴ and R ⁴ , where the radicals R ⁴ , R ⁴ and R ⁶ are selected independently of one another according to the meanings given below,

A is a group selected from the group comprising Ci.-io

Alkanediyl, C _2- ιo-alkenediyl, C _2- ιo alkynediyl and (co _-3 alkanediyl-C _3-7 _{cycloalkanediyl-co-3} alkanediyl), wherein in a five-membered cycloalkyl ring one ring member ring N or ring May be 0 and in a six- or seven-membered cycloalkyl ring one or two ring members may each be ring N and / or ring O atoms, wherein the ring N atoms optionally with C _1-3 alkyl or -C _-3 Alkanoyl may be substituted, wherein in the abovementioned aliphatic chains, a C atom can be exchanged for O, NH, N-Cι- ₃ alkyl or Nd _-3 alkanoyl and wherein alkyl or cycloalkyl optionally with a radical selected from the group comprising = 0, OH, 0-Cι- _3- alkyl, NH _2l NH-C _1-3 -alkyl, NH-C ₃ -alkanoyl, N (Cι- _3- alkyl) ₂ and N (C _-3 -

Alkyl) (C _1-3 alkanoyl), may be substituted, B is a radical selected from the group comprising COOH, COOR ⁵ ,

CONH ₂ , CONHNH2, CONH? ⁵ , CONR ⁵ R ^{5 '} , CONHOH, CONHOR ⁵ and

Tetrazolyl, each bonded to a C atom of group A, wherein the radicals R ⁵ and R ^{5 are} independently selected according to the meanings given below,

Y is a group selected from the group comprising O, NH,

where R ⁴ and R ^{6 have} the meanings given below,

wherein in the above radicals R ⁴ , R ⁴ , R ⁵ , R ⁵ and R ^{6 have} the following meanings; in it are:

R ⁴ and R ⁴ are each independently a radical selected from among

Group, _CF3, C2F5, Cι _-4 alkyl, C comprising _2-4 alkenyl, C _2-3 alkynyl and (C _0- 3-alkanediyl-C _3-7 cycloalkyl), wherein, in a five-membered cycloalkyl ring Ring member ring-N or ring-0 can be and in a six- or seven-membered cycloalkyl ring one or two ring members each ring N and / or ring O atoms may be, wherein the ring N-atoms optionally with -C _-3 alkyl or Cι _-3- alkanoyl may be substituted,

R ⁵ and R ⁵ 'are each independently a radical selected from among

Group comprising Cι _-6 alkyl, C2-6 alkenyl, C _2- 6 -alkynyl, where one C atom to O, S, SO, S0 _2, NH, N-alkyl or Cι _-3 NC _{1- 3-} alkanoyloxy, furthermore (co-3-alkanediyl-C _3-7 -cycloalkyl), wherein in a five-membered cycloalkyl ring, a ring member ring-N or ring-0 may be and in a six- or seven-membered Cycloalkyl ring may be one or two ring members each ring N and / or ring O atoms, wherein the ring N atoms may optionally be substituted by -CC 3-alkyl or C ₃ alkanoyl, and also (C _{0 -} 3-alkanediyl-aryl) and (C _0-3 -alkanediyl-heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, all of the foregoing Alkyl and cycloalkyl radicals having up to two radicals selected from the group comprising CF ₃ , C ₂ F ₅ , OH, 0-C _1-3 alkyl, NH ₂ , NH-C _1-3 alkyl, NH-Cι _3- alkanoyl, N (C _1-3

Alkyl) ₂ , N (C _1-3 alkyl) (C _1-3 alkanoyl), COOH, CONH ₂ and COO-Cι _-3 - alkyl, and all of the aforementioned aryl and heteroaryl groups having up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and SO ₂ NH ₂ can be and / or a fancied

Or R ⁵ and R ⁵ together with the amide N atom of B form a five- to seven-membered, saturated or unsaturated heterocyclic ring which is another N- or O- or may contain S atom and which may be substituted by C _1-4 alkyl,

(C _0- alkanediyl 2-Cι _-4 alkoxy), C-ι- -alkoxycarbonyl, aminocarbonyl or aryl,

a radical selected from the group comprising (C _0-3 alkanediyl-aryl) and (C _0-3 -alkanediyl-heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, and wherein the aryl and heteroaryl groups may be substituted with up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH 3, OC ₂ H ₅ , N0 ₂ , N (CH ₃ ) ₂ , CF _3> C ₂ F ₅ and S0 ₂ NH ₂ , and / or may also carry a fused methanediylbisoxy or ethane-1, 2-diylbisoxy group, R ⁷ and R ^{7 'are} independently R ⁴ or R ⁶ .

Preferred are those benzimidazole derivatives in which the substituent B-A-Y is bonded to the 6-position of the benzimidazole.

The present invention also includes physiologically acceptable salts and esters of the abovementioned compounds, in particular the acid salts of the nitrogen bases of the benzimidazole derivatives according to the invention, furthermore the salts of carboxylic acids of the derivatives according to the invention with bases and the esters of the carboxylic acids of the derivatives and of carboxylic acids derived from carboxylic acid derivatives are, for example of Carbsonäureamiden.

The benzimidazole derivatives may have one or more chiral centers such that the compounds may exist in several isomeric forms. The compounds of the formula I can also be present as tautomers, stereoisomers or geometric isomers. The invention also includes all possible isomers, such as E and Z isomers, S and R enantiomers, diastereomers, racemates and mixtures thereof, including the tautomeric compounds. All these isomeric compounds are - even if not expressly stated - part of the present invention. The isomer mixtures can be separated into the enantiomers or E / Z isomers by customary methods, for example crystallization, chromatography or salt formation.

The heteroaryl groups contained in the benzimidazole compounds are composed of five or six skeleton atoms and may contain one or two heteroatoms. Heteroatoms are oxygen (O), nitrogen (N) and sulfur (S). Examples of heteroaryl groups are pyrrolyl, thienyl, furanyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl and pyridazinyl. If the heteroaryl groups are part of R ¹ or R ² , the group is bonded via a C atom to the respective N atom of the benzimidazole skeleton or to the substituent Z. Suitable aryl radicals are, in particular, the phenyl radical, but also the naphthyl radical. The aryl and heteroaryl radicals can be attached in any manner to the benzimidazole skeleton or another group, for example as 1- or 2-naphthyl or as 2-, 3- or 4-pyridinyl.

Alkyl groups can be straight-chain or branched. Examples of alkyl groups are methyl, ethyl, π-propyl, / ^'so-propyl, n-butyl, s /-butyl, tert-butyl, n-pentyl, s / -pentyl, erf-pentyl, neo-pentyl, π- Hexyl, sec-hexyl, heptyl, octyl, nonyl, decyl. Also, the higher homologues comprise both the linear and the branched alkyl groups, that is, for example, 2-ethylhexyl for octyl and 3-propyl-hexyl for nonyl.

Perfluorinated alkyls are preferably CF ₃ and C ₂ F ₅ .

Alkenyl groups can be straight-chain or branched. Examples are vinyl, 2-propenyl, 1-propenyl, 2-butenyl, 1-butenyl, 1-methyl-1-propenyl, 2-methyl-2-propenyl and 3-methyl-2-propenyl Akenylreste in the context of the invention.

Alkynyl groups can be straight-chain or branched. Examples of these are ethynyl, 1-propynyl, 2-propynyl, 1-butynyl and 2-butynyl.

Cycloalkyl groups are in each case preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl (corresponds to C _3-7 -cycloalkyl).

Examples of saturated heterocyclic rings or cycloalkyls containing one or more heteroatoms are: piperidine, pyrrolidine, tetrahydrofuran, morpholine, piperazine, hexahydroazepine and 2,6-dimethylmorpholine, N-phenylpiperazine, methoxymethylpyrrolidine, where the

Linkage with a C-atom adjacent to the ring can take place via optionally present ring N atoms. Alkanediyl, alkenediyl, alkynediyl and cycloalkanediyl radicals mentioned in the description of the invention are synonymous with alkylene, alkenylene, alkynylene and cycloalkylene. Insofar as the number of C atoms contained in the general formulas of the alkanediyl radicals is given and the lower limit of this number is given as the value 0, this alkanediyl radical is not included in the respective case.

Examples of alkanes, alkenes and alkynes for A are: straight-chain or branched alkanediyl having one to eight carbon atoms, for example methanediyl, ethanediyl, propanediyl, butanediyl, pentanediyl, hexanediyl, furthermore 1-methylethanediyl, 1-ethylethanediyl, 1-methylpropanediyl, 2-methylpropanediyl, 1-methylbutanediyl, 2-methylbutanediyl, 1-ethylbutanediyl, 2-ethylbutanediyl, 1-methylpentanediyl, 2-methylpentanediyl, 3-methylpentanediyl and analogous compounds.

Straight-chain or branched alkenediyl and alkynediyl having two to eight carbon atoms are alkenediyl groups or alkynediyl groups having double and triple bonds in all possible positions and with all possible methyl or ethyl substitutions. In these radicals in each case one or two C-atoms by O, NH, N-alkyl or N-Cι _-3 _-3 Cι can alkanoyl be exchanged, wherein the replaced group is separated by at least two C-atoms of V.

If two radicals are ortho-stationary, they can form a common ring with the adjacent aromatic. Compounds in which N, O or S atoms are bonded to olefinic or acetylenic multiple bonds, or in which a plurality of N, O, S or halogen atoms are bonded to the same aliphatic C atom, or in which N, O or S atoms are bonded to the same aliphatic C atom. , O- or S-atoms are bonded directly to each other are excluded, unless these links are not explicitly defined, for example in the recited in the claim functional groups or in heteroaromatics. The physiologically acceptable acid salts of the nitrogen bases of the benzimidazole derivatives according to the invention can be formed with inorganic and organic acids, for example with oxalic acid, lactic acid, citric acid, fumaric acid, acetic acid, maleic acid, tartaric acid, phosphoric acid, hydrochloric acid, hydrobromic acid, sulfuric acid, p-toluenesulfonic acid and methanesulfonic acid.

For salt formation of acid groups, in particular carboxylic acid groups, also the inorganic or organic bases are suitable, which are known for the formation of physiologically acceptable salts, such as, for example

Alkali hydroxides, in particular sodium and potassium hydroxide, alkaline earth hydroxides, such as calcium hydroxide, also ammonia, and amines, such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and tris (hydroxymethyl) - methylamine.

For Esterbildung are all monohydric lower, divalent and trivalent alcohols suitable, in particular methanol, ethanol, / ^'so-propanol and tert-butanol, and ethylene glycol and glycerol.

Particular preference is given to benzimidazoles of the general formula I in which the radicals and groups given below have the following meanings, independently of one another:

R ^{1 is} a phenyl group which may be substituted with up to two independently selected radicals selected from the group comprising:

F, Cl, Br,

C (NH) NH ₂ , C (NH) NHR ⁴ , C (NH) NR ⁴ R ⁴ ', C (NR ⁴ ) NH ₂ , C (NR ⁴ ) NHR ^4' ,

C (NR ⁴ ) NR ⁴ R ⁴ ', OH, OR ⁴ , OCOR ⁴ , OCONHR ⁴ ,

COR ⁴ , C (NOH) R ⁴ ,

CN, COOH, COOR ⁴ , CONH ₂ , CONRV ^' , CONH ⁴ , CONHOH,

SR ⁴ , SOR ⁴ , SO ⁴ , SO ₂ NH ₂ , SO ₂ NHR ⁴ , SO ₂ NR ⁴ R ^{4 '} , NO ₂ , NH ₂ , NHR ⁴ , NRV, NHCONHR ⁴ and

R ⁴ , wherein the radicals R ⁴ and R ⁴ are selected independently of one another according to the meanings given below, and where two substituents on R ¹ can be linked to one another such that they together form a methanediylbisoxy, ethane-1, 2-diylbisoxy, propane Form 1, 3-diyl or butane-1, 4-diyl group, if they are ortho on each other,

R ^{2 is} a mono- or bicyclic Cβ-io-aryl group or a mono- or bicyclic 5-10-membered heteroaryl group having 1-2 heteroatoms selected from the group consisting of N, S or O, said aryl or heteroaryl group with up to three of the following substituents independently of one another: F, Cl, Br,

XOH, XOR ⁴ , XOCOR ⁴ , XOCONHR ⁴ , XOCOOR ⁴ , XCOR ⁴ , XC (NOH) R ⁴ , XC (NOR ⁴ ) R ⁴ , XC (NO (COR ⁴ )) R ⁴ , XCOOH, XCOOR ⁴ , XCONH ₂ , XCONHR ⁴ , XCONR ⁴ R ⁴ , XCONHOH,

XCONHOR ⁴ , XCOSR ⁴ ,

XSR ^4, XSOR ^4, XS0 ₂ R ^4, S0 ₂ NH _2, S0 ₂ NHR ^4, S0 ₂ NR ⁴ R ^4, N0 _2, XNHR ^4, XNR ⁴ R ^4, XNHS0 ₂ R ^4, XN (S0 ₂ R ⁴ ) S0 ₂ R ⁴ , XNR ⁴ S0 ₂ R ⁴ , R ⁴ , where two substituents on R ² , when ortho to each other, can be linked to each other in such a way that together they are methanediylbisoxy, ethane-1,2-diylbisoxy, propane 1, 3-diyl, butane-1, 4-diyl,

R ^{3 is} a radical selected from the group comprising hydrogen, F,

Cl, Br, CH ₃ , C ₂ H ₅ , CF ₃ , C ₂ F ₅ , OH, OR ⁴ , NHSO ₂ R ⁶ and NHCOR ⁴ , where R ⁴ and R ^{6 have} the meanings given below,

A Cι _-10- alkanediyl, C _2- ιo-alkenediyl, C _2- ιo-alkinediyl, (C _0-5 -alkanediyl-C _3-7 - cycloalkanediyl-Co- ₅ alkanediyl), wherein in a 5-membered cycloalkyl ring a ring member may be an N or an O and in a 6- or 7-membered cycloalkyl ring may be one or two ring members N and / or O, wherein ring nitrogens optionally with C ^ -Alky! or Cι- ₃ alkanoyl may be substituted, wherein, in the above-mentioned aliphatic chains one carbon atom or two carbon atoms for O, NH, NCι _-3 - alkyl, alkanoyl NCι- ₃ may be replaced,

B is a radical selected from the group comprising COOH, COOR ⁵ ,

CONH _2> CONHR ⁵ and CONR ⁵ R ^{5 '} , each attached to a C atom of group A, wwoobbeeii ddiiee RReesste R ⁵ and R ⁵ are selected independently of one another according to the meanings given below,

O

wherein in the above radicals R ⁴ , R ⁴ , R ⁵ , R ⁵ and R ^{6 have} the following meanings; in it mean:

R ⁴ and R ^{4 are the} same as stated above,

R ⁵ and R ⁵ each independently represent a radical selected from the group consisting of Cι _-6 alkyl, C _2- 6 alkenyl, C _2- 6-alkynyl, where a

C-atom is replaced by O, S, SO, S0 _2, NH, N-Cι _-3 alkyl or NC _1-3 alkanoyl may be exchanged, also (C _0- 3-alkanediyl-C _3- 7 cycloalkyl), wherein in a five-membered cycloalkyl ring, a ring member Ring-N or Ring-0 may be and in a six- or seven-membered cycloalkyl ring one or two ring members may each be ring N and / or ring O atoms, wherein the ring N-atoms optionally with Cι- ₃ alkyl or -C _3- alkanoyl, as well as (C _0-3 alkanediyl-phenyl) and

(Co- _3- alkanediyl-heteroaryl), wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, wherein all the aforementioned alkyl and cycloalkyl radicals are selected with a radical from the group comprising CF ₃ , C ₂ F ₅ , OH, OC ₁ -

₃ alkyl, NH _2, NH-Cι _-3 alkyl, NH-C _1-3 -alkanoyl, N (Cι _-3 alkyl) _2, N (C _{1 -3} - alkyl) (Cι _-3 alkanoyl) , COOH, CONH ₂ and COO-C ₃ alkyl, and all of the aforementioned phenyl and heteroaryl groups having up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H 5, NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , and / or can also carry a fused methanediylbisoxy or ethane-1,2-diylbisoxy group, or R ⁵ and R ⁵ together with the amide N atom of B form a five- to seven-membered, saturated or unsaturated heterocyclic ring which forms another N- or O- or S-

May contain atom and which may be substituted by Cι _-4 alkyl, (C _0-2 alkanediyl-Cι- alkoxy), Cι- alkoxycarbonyl, aminocarbonyl or phenyl,

a phenyl or heteroaryl group wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, and wherein the phenyl and heteroaryl groups may be substituted with up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH 3, OC ₂ H ₅ , NO ₂₎ N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and

SO ₂ NH ₂ , or else a fused methanediylbisoxy or ethane-1, 2-diylbisoxygruppe can carry. R ³ is preferably hydrogen. The group YA is represented in a preferred embodiment by a C _1-6 alkyleneoxy group which is bonded via the O atom to the benzimidazole skeleton.

Furthermore, the invention relates to pharmaceutical compositions containing one or more compounds of the general formula I and one or more excipients. The pharmaceutical compositions or compositions of the invention are prepared with conventional solid or liquid carriers or diluents and customary pharmaceutical and technical excipients according to the desired mode of administration with a suitable dosage in a manner known per se. Preferred preparations consist of a dosage form which is suitable for oral, enteral or parenteral, for example i.p. (intraperitoneal), i.v. (intravenous), i.m. (intramuscular), or percutaneous, application is appropriate. Such dosage forms are, for example, tablets, film-coated tablets, dragees, pills, capsules, powders, creams, ointments, lotions, liquids, such as syrups, gels, injectable liquids, for example for ip, iV, im, or percutaneous injection, etc. Furthermore, depot forms, as implantable preparations, and suppositories suitable. Depending on their nature, the individual preparations give the derivatives according to the invention gradually or the entire amount to the body in a short time.

For oral administration, capsules, pills, tablets, dragees and liquids or other known oral dosage forms may be used as pharmaceutical preparations. In this case, the drugs may be formulated to either release the drugs in a short time and deliver them to the body or have a depot effect, so that a longer-lasting, slow supply of active ingredient to the body is achieved. The dosage units may contain, in addition to the at least one benzimidazole derivative one or more pharmaceutically acceptable carriers, for example rheology of the drug, surfactants, solubilizers, microcapsules, microparticles, granules, thinners, binders such as starch, sugar, sorbitol and Gelatin, further fillers such as silica and talc, lubricants, dyes, fragrances and other substances.

Corresponding tablets can be prepared, for example, by mixing the active compound with known excipients, for example inert diluents such as dextrose, sugar, sorbitol, mannitol, polyvinylpyrrolidone, disintegrants such as corn starch or alginic acid, binders such as starch or gelatin, lubricants such as carboxypolymethylene, carboxymethylcellulose, cellulose acetate phthalate or polyvinyl acetate , to be obtained. The tablets can also consist of several layers.

Coated tablets can accordingly be produced by coating cores produced analogously to the tablets with agents customarily used in tablet coatings, for example polyvinylpyrrolidone or shellac, gum arabic, talc, titanium oxide or sugar. In this case, the dragee wrapper can also consist of several layers, wherein the auxiliaries mentioned above in the case of the tablets can be used.

Active ingredients containing capsules can be prepared, for example, by mixing the active ingredient with an inert carrier such as lactose or sorbitol and encapsulated in gelatin capsules.

The active compounds may also be formulated in the form of a solution intended for oral administration which, in addition to the active benzimidazole derivative, contains as ingredients a pharmaceutically acceptable oil and / or a pharmaceutically acceptable lipophilic surfactant and / or a pharmaceutically acceptable hydrophilic, surfactant and / or a pharmaceutically acceptable water-miscible solvent.

In order to achieve a better bioavailability of the active compounds, the compounds can also be formulated as Cyclodextrinchlatrate. To do this the compounds reacted with α-, ß- or γ-cyclodextrin or derivatives thereof.

If creams, ointments, lotions and externally applicable liquids are to be used, they must be such that the

Compounds are supplied to the body in sufficient quantity. In these dosage forms, adjuvants are included, such as drugs to adjust the rheology of the drugs, surfactants, preservatives, solubilizers, thinners, substances to increase the permeability of the active ingredients through the skin, dyes, fragrances and skin protection agents, such as conditioners and moisture regulators. Other active ingredients may also be present in the drug together with the compounds [Ullmanns Enzyklopadie der technischen Chemie, Vol. 4 (1953), pages 1-39; J. Pharm. Sci., 52, 918 et seq. (1963); H. v. Czetsch-Lindenwald, adjuvants for pharmacy and adjacent areas; Pharm. Ind., 2, 72 ff. (1961); Dr. H. P. Fiedler, Lexicon of adjuvants for pharmacy, cosmetics and adjacent areas, Cantor AG, Aulendorf / Württ., 1971].

The substances can also in suitable solutions such as saline, as an infusion or injection solution for

Application come. For parenteral administration, the active ingredients may be dissolved or suspended in a physiologically acceptable diluent. Particularly suitable diluents are oily solutions, for example solutions in sesame oil, castor oil and cottonseed oil. To increase solubility, solubilizers, such as benzyl benzoate or benzyl alcohol, may be added.

For the formulation of an injectable preparation, any liquid carrier can be used in which the compounds according to the invention are dissolved or emulsified. These liquids often also contain substances for the regulation of viscosity, surfactants, preservatives, solubilizers, thinners and other additives with which the solution isotonic. Other active ingredients can also be administered together with the benzimidazole derivatives.

It is also possible to incorporate the substances into a transdermal system and thus transdermally apply them. For this purpose, the benzimidazole derivatives are applied in the form of a depot injection or an implant preparation, for example subcutaneously. Such preparations may be formulated to allow sustained release of the active ingredient. For this purpose, known techniques can be used, for example, dissolving or working with a membrane depots. Implants may contain as inert materials, for example biodegradable polymers or synthetic silicones, for example silicone rubber. The benzimidazole derivatives may also be incorporated into a patch for percutaneous administration, for example.

The dosage of the substances will be determined by the attending physician and depends inter alia on the substance administered, the route of administration, the disease to be treated and the severity of the disease. The daily dose is not more than 1000 mg, preferably not more than 100 mg, which dose may be given as a single dose to be administered once or divided into two or more daily doses.

Investigation of biological properties

Example 1: Activation of Macrophages

To test the substances on macrophages / monocytes, LPS-activated THP-1 cells were used. For this purpose, 2.5 × 10 6 cells / ml were seeded in RPMI medium (RPMI 1640 + 10% FCS). The compounds of the invention were added at a concentration of 5 μM and preincubated for 30 minutes. The cells were stimulated overnight at 37 ° C with 1 μg / ml LPS. Thereafter, the medium was harvested and the amount of TNFα was quantified. The treatment of the cells with the substances according to the invention resulted in a reduction of the TNFα amount of at least 30%.

Example 2:

Inhibition of TNFα and IL 12 production in THP-1 cells

The inhibition of cytokine production can be determined, for example, by measuring

TNFα and interleukin 12 in lipopolysaccharide (LPS) -stimulated THP-1 cells.

To this is added 2.5 x 10 ⁶ cells of THP-1 (American Type Culture Company, Rockville, Md) / ml of RPMI 1640 Medium (Life Technologies) / 10% FCS (Life Technologies, Cat # 10270-106) 96 well flat-bottom cell culture plates (TPP, Product No. 9296) seeded (100 μl / well). The compounds of the invention are added at different concentrations and preincubated for 30 minutes. The predilution of the test substances was carried out in the incubation medium. The addition of the test substances takes place as a doubly concentrated substance solution (100 μl / well). Cells were stimulated overnight at 37 ° C with 0.1 μg / ml LPS (Sigma L2630, from E. Coli serotype 0111. B4). Thereafter, the medium was harvested and the amount of TNFα or Interleukin 12 quantified. To measure TNFα, a commercially available TNFα kit from CIS bio international was used (product no. 62TNFPEB). The interleukin 12 amount was determined using the ORIGEN technology (IGEN International, Inc., Gaithersburg, Maryland). The calculated IC50 value corresponds to the concentration of test substance needed to achieve a 50% inhibition of maximal TNFα or interleukin 12 production. The substances show an IC50 below 10 μM in this experiment. With the help of similar methods, the inhibition of IL 12 and TNFα with the substances can also be demonstrated with the use of peripheral blood leukocytes and comparable stimuli.

Example 3:

Inhibition of IFNγ production by peripheral blood mononuclear cells

To demonstrate the influence of the substances on the T cell activation, for example, the measurement of IFNγ secretion can be used. Human whole blood was used to isolate peripheral mononuclear cells (blood collection via Na citrate S monovettes "Coagulation 9 NC / 10 ml" / Sarstedt.) Enrichment of the blood cells was carried out by density gradient centrifugation: 15 ml of Histopaque 1077 (Sigma, Cat No H8880) in LEUCOSEP tubes (Greiner, cat # 227290) and centrifuged for 30 seconds at 1000 g, then 15 ml of whole blood are added and centrifuged for 10 minutes at 1000 g.Finally, the upper plasma layer Pipette and transfer the underlying cell layer (peripheral blood mononuclear cells) into 15 ml sample tubes (Falcon) and then wash several times with 10 ml HBSS (HANKS balanced solution (without Mg2 + and Ca2 +), cat no 14175-53) Cell pellet in culture medium RPMI 1640 + 25 mM Hepes (Life Technologies Cat. No. 52400-041 10% FCS (Life Technologies, cat. No. 10270-106), 0.4% penicillin-streptomycin solution g (Life Technologies, cat. no. 15140-106) (1 x 10 ⁶ cells / ml). Each 100 .mu.l cell suspension solution was distributed to 96-well flat-bottomed cell culture plates (TPP, product no. 9296) and stimulated with 5 ug / ml phytohemagglutinin. The substances according to the invention were added in different concentrations and preincubated for 30 minutes. The stimulation of the cells took place over a period of 48 h. Thereafter, the medium was harvested and the amount of IFNγ was quantified. The amount of IFNγ was determined by ORIGEN technology (IGEN International, Inc., Gaithersburg, Maryland). The calculated IC50 value corresponds to the concentration of test substance needed to achieve a 50% inhibition of maximum IFNγ production.

Treatment of the cells with the test substances resulted in a reduction in the amount of IFNγ of at least 30% at a concentration of 10 μM. With the help of a similar method can also with the use of specific T-cell activators, such as. As monoclonal anti-CD3 antibody, the inhibition of IFNγ are shown with the substances.

Tab. 1

Substance 1: 6 - [[1- (4-methylphenyl) -2-phenyl-1H-benzimidazol-6-yl] oxy] hexanoic acid isopropyl ester

Substance 2:

6 - [[1- (4-methylphenyl) -2-phenyl-1H-benzimidazol-6-yl] oxy] hexanoic acid

Example 4:

Induction of IL-10 Production of Peripheral Blood Mononuclear Cells Induction of IL-10 production is exemplified by measurement of IL-10 in phytohemagglutinin (PHA) or lipopolysaccharide (LPS) stimulated peripheral blood mononuclear cells. For the isolation of peripheral blood mononuclear cells, whole human blood was used (blood collection via Na citrate S monovettes "Coagulation 9 NC / 10 ml7 Sarstedt). Enrichment of the lymphocytes and monocytes was carried out by density gradient centrifugation. 15 ml of Histopaque-1077 (Sigma, cat. No. H8880) were introduced into 50 ml of LEUCOSEP tubes (Greiner, cat. No. 227290) and centrifuged for 30 seconds at 250g pushed down through the frit in the tube. Subsequently, 20 ml of whole blood are added and centrifuged for 15 min. At 800 g and room temperature. After centrifugation, the supernatant (plasma and platelets) is pipetted off and discarded and the underlying cell layer (lymphocytes and monocytes) transferred to 50 ml centrifuge tubes (Falcon) and then washed 3 times in culture medium VLE RPMI 1640 (Seromed, No. FG1415) (centrifugation 10 min. each at 250g, room temperature). Finally, the cell pellet is incubated in culture medium VLE RPMI 1640 (Seromed, No. FG1415), 10% FCS (Life Technologies, cat. No. 16000-044, low endotoxin, heat-inactivated 1 h, 56 ° C), 50 μg / ml Penicillin-streptomycin solution (Life Technologies, cat. No. 15140-106) and after cell counting by trypan blue staining set to 3 x 10 ⁶ cells / ml. Each 100 μl of cell suspension solution was distributed to 96-well flat-bottomed cell culture plates (Costar, product no. 3599). In each case 100 μl of 3 × concentrated stimulation solution (3 μg / ml LPS of E. coli serotype 0127: B8, Sigma, Cat.No. L-4516 or 300 μg / ml PHA-L, Biochrom KG, Cat. M5030). The substances according to the invention were added in different concentrations as a 3-fold concentrated substance solution (100 μl / well). The stimulation of the cells was carried out at 37 ° C and 5% C0 ₂ over a period of 24 h. Thereafter, the cell culture supernatant was harvested and IL-10 quantified. The IL-10 concentration was determined using a commercially available ELISA kit from BioSource International (Human IL-10, cat.No .: KHC0101C). The calculated EC ₅₀ value corresponds to the concentration of test substance needed to increase IL-10 secretion by 50% of the maximum increase.

The compounds of the invention increase the IL-10 production of peripheral blood mononuclear cells. Example 5:

Inhibition of TNFα and IL-12 HD Production of Peripheral Blood Mononuclear Cells The inhibition of TNFα and IL-12 HD p70 production is demonstrated, for example, by measuring TNFα and IL-12 HD p70 in with lipopolysaccharide (LPS) and interferon gamma (IFNγ). shown stimulated peripheral blood mononuclear cells. For the isolation of peripheral blood mononuclear cells, whole human blood was used (blood collection via Na citrate S monovettes "Coagulation 9 NC / 10 ml7 Sarstedt). Enrichment of the lymphocytes and monocytes was carried out by density gradient centrifugation. 15 ml of Histopaque-1077 (Sigma, cat. No. H8880) were introduced into 50 ml of LEUCOSEP tubes (Greiner, cat. No. 227290) and centrifuged for 30 seconds at 250g pushed down through the frit in the tube. Subsequently, 20 ml of whole blood are added and centrifuged for 15 min. At 800 g and room temperature. After centrifugation, the supernatant (plasma and platelets) is pipetted off and discarded and the underlying cell layer (lymphocytes and monocytes) transferred to 50 ml centrifuge tubes (Falcon) and then washed 3 times in culture medium VLE RPMI 1640 (Seromed, No. FG1415) (centrifugation 10 min. each at 250g, room temperature). Finally, the cell pellet is incubated in culture medium VLE RPMI 1640 (Seromed, No. FG1415), 10% FCS (Life Technologies, cat. No. 16000-044, low endotoxin, heat-inactivated 1 h, 56 ° C), 50 μg / ml Penicillin-streptomycin solution (Life Technologies, cat. No. 15140-106) and after cell counting by trypan blue staining set to 3 x 10 ⁶ cells / ml. Each 100 μl of cell suspension solution was distributed to 96-well flat-bottomed cell culture plates (Costar, product no. 3599). To this was added in each case 100 μl of 3 × concentrated stimulation solution (3 μg / ml LPS of E. coli serotype 0127: B8, Sigma, cat No. L-4516 and 30 ng / ml IFNγ 1 b, Imukin, Boehringer Ingelheim). The substances according to the invention were added in different concentrations as a 3-fold concentrated substance solution (100 μl / well). The cells were stimulated at 37 ° C and 5% C0 ₂ over a period of 24 h. Thereafter, the cell culture supernatant was harvested and the concentrations of TNFα and IL-12 HD p70 using commercially available ELISA kits from BioSource International (TNF-α EASIA, cat.

No. KAC1752) and R & D Systems (Quantikine ™ HS IL-12, Cat. No. HS 120).

The calculated IC ₅₀ value corresponds to the concentration of test substance that is needed to prevent a 50% inhibition of the maximum TNFα or

Interleukin 12 HD p70 production.

The compounds according to the invention inhibit TNFα and IL-12 HD p70

Production of peripheral blood mononuclear cells.

Tab. 2

Substance 2: 6 - [[1- (4-methylphenyl) -2-phenyl-1H-benzimidazol-6-yl] oxy] hexanoic acid

Claims

claims

1 . Use of a microglial inhibitor for the manufacture of a medicament for the treatment of monocyte, macrophage or T cell mediated immune responses.

2. Use according to claim 1 for the treatment of interleukin 12 (IL 12) and interferon γ (IFNy) -mediated immune reactions.

3. Use according to claim 1 for the treatment of non-T-cell-mediated inflammatory reactions.

4. Use according to claim 1 for the treatment of autoimmune diseases, inflammatory diseases that are not based on neuroinflammation, allergic and infectious diseases.

5. Use according to claim 4 for the treatment of chronic inflammatory bowel diseases, for example inflammatory bowel diseases, Crohn's disease, or ulcerative colitis, arthritis, allergic contact dermatitis, psoriasis, pemphigus, asthma, diabetes, type I insulin-dependent diabetes mellitus, rheumatoid arthritis, Lupus Diseases and Other Collagenosis, Graves' Disease, Hashimoto's Disease, Graft versus Host Disease and Transplant Disease, Sarcoidosis, Asthma, Hypersensitive Pneumonitis, Sepsis, Septic Shock, Endotoxin Shock, Toxic Shock Syndrome, Toxic Liver Failure, ARDS (Acute

Respiratory distress syndrome), eclampsia, cachexia, acute viral infections, organ damage after reperfusion, "first dose response" after administration of anti-T cell antibodies.

6. Use of a benzimidazole of the formula I, its tautomeric or isomeric form or salt

wherein

R ^{1 is} an aryl group or a five- or six-membered one

F, Cl, Br,

C (NH) NH ₂ , C (NH) NHR ⁴ , C (NH) NRV ^' , C (NR ⁴ ) NH ₂ , C (NR ⁴ ) NHR ^4' ,

C (NR ⁴ ) NR ⁴ R ^{4 '} ,

X-OH, X-OR ⁴ , X-OCOR ⁴ , X-OCONHR,

X-COR ⁴ , XC (NOH) R ⁴ ,

X-CN, X-COOH, X-COOR ⁴ , X-CONH ₂ , X-CONR ⁴ R ^{4 '} , X-CONHR ⁴ ,

X-CONHOH,

X-SR ⁴ , X-SOR ⁴ , X-SO ² R ⁴ ,

SO ₂ NH 2, S0 ₂ NHR ^4, S0 ₂ NR ⁴ R ^{4 ',}

N0 _2, X-NH _2l X-NHR ^4, X-NRV ^', X-NHSO2R ^4, X-NR ⁴ S0 ₂ R ^4',

X-NHCOR ⁴ , X-NHCOOR ⁴ , X-NHCONHR ⁴ and

R ⁴ , wherein X is a bond, CH ₂ , (CH ₂ ) ₂ or CH (CH ₃ ) ₂ , wherein furthermore the radicals R ⁴ and R ⁴ are selected independently of one another according to the meanings given below, and wherein two substituents on R ¹ , when ortho to each other, may each be linked to each other so as to form together a methanediylbisoxy, ethane-1,2-diylbisoxy, propane-1,3-diyl or butane-1,4 form diyl group, or

a radical selected from the group consisting of Cι _-6 alkyl, (C _0- 3-alkanediyl-C _3- 7 cycloalkyl) C _3- and 6Alkenyl, in which a hydrogen atom is replaced by a heterocyclic radical selected from the group compound comprising piperazine, morpholine, piperidine and pyrrolidine, which may be exchanged to form a bond to a first N-atom of the heterocyclic radical, wherein said alkyl, cycloalkyl, alkenyl and heterocyclic radicals are substituted with up to two radicals selected from the group comprising C _0-2 alkanediyl-OH, C _0-2 alkanediyl-OR ⁷ ,

C _{_0-2} alkanediyl-NH _2, C _0-2 alkanediyl-NHR ^7, C _0-2 alkanediyl-NR ⁷ R ^{7 ',} Co-2-alkanediyl NHCOR ^7, C _0- 2-alkanediyl-NR ⁷ COR ⁷ ', C ₀ . _2- alkanediyl-NHSO ₂ R ⁷ , C _0-2 -alkanediyl-NR ⁷ SO ₂ R ^{7 '} , C _0-2 -alkanediyl-CO ₂ H, C _0-2 - alkanediyl-C0 ₂ R ⁷ , C _0- 2-alkanediyl-CONH _2, C _0- alkanediyl 2-CONHR ⁷ C _0- alkanediyl 2

CONR ⁷ R ⁷ , phenyl and a five- or six-membered heteroaryl radical, wherein the heteroaryl radical contains one or two heteroatoms selected from the group comprising N, S and O, where furthermore the phenyl and the heteroaryl radical having up to two radicals, selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH,

OCH3, OC2H5, N0 _2, N (CH ₃₎ _2, CF _3, C ₂ F ₅ and S0 ₂ NH _2, may be substituted and / or 2-diylbisoxygruppe also carry a fused Methandiylbisoxy- or ethane-1, wherein the piperazine radical on a second nitrogen atom can also be substituted by R ⁷ , COR ⁷ or SO ₂ R ⁷ , where R ⁷ and R ⁷ 'can be selected independently of one another in accordance with the meanings given below, R ² -ZR ² 'is an aryl group or a five- or six-membered one

F, Cl, Br,

C (NH) NH ₂ , C (NH) NHR ⁴ , C (NH) NRV ', C (NR ⁴ ) NH ₂ , C (NR ⁴ ) NHR ^4' ,

C (NR ⁴ ) NR ⁴ R ^{4 '} , X-OH, X-OR ⁴ , X-OCOR ⁴ , X-OCONHR ⁴ ,

X-COR ⁴ , XC (NOH) R ⁴ ,

X-CN, X-COOH, X-COOR ⁴ , X-CONH ₂ , X-CONRV, X-CONHR ⁴ ,

X-CONHOH,

N0 ₂ , X-NH ₂ , X-NHR ⁴ , X-NR ⁴ R ^{4 '} , X-NHSO ² R ⁴ , X-NR ^ OsR ^4' ,

X-NHCOR ⁴ , X-NHCOOR ⁴ , X-NHCONHR ⁴ and

R ⁴ , wherein X is a bond, CH ₂ , (CH ₂ ) ₂ , or CH (CH ₃ ) ₂ , further wherein the radicals R ⁴ and R ⁴ 'are independently selected according to the meanings given below and wherein two radicals R ¹ , when ortho to each other, may be linked together to form together a methanediylbisoxy, ethane-1,2-diylbisoxy, propane-1,3-diyl or

Butane-1, 4-diyl group,

Z is NH, NR ^{2 "} , O, S, SO or S0 ₂ , where R ^{" has} the meaning given below,

R ² and R ² are each independently a radical selected from the group comprising: C _1-4 perfluoroalkyl, C _1-6 alkyl, (C ₀ -3 alkanediyl C _3-7 cycloalkyl), (Co- ₃ -alkanediyl-aryl) and (C ₀ -3 alkanediyl-heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two hetero atoms selected from the group comprising N, S and O, and wherein the aryl and heteroaryl group each having up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH _2> may be substituted and / or may also carry a fused methanediylbisoxy or ethane-1, 2-diylbisoxygruppe, and further a ring member in a five-membered cycloalkyl Ring-N or ring can be 0 and one or two ring members in a six- or seven-membered cycloalkyl ring N and / or ring O atoms may be, where the ring N atoms optionally substituted by Cι _-3 alkyl or Cι _-3- alkanoyl R ² and R ² together with Z form a five- to seven-membered heterocyclic ring when Z is an N atom, wherein Z is the further indicated above, further wherein the heterocyclic ring containing another N, O or S atom and may be optionally substituted with a radical selected from the group comprising Cι- alkyl, (C _0-3 alkanediyl -C _1-3 alkoxy), Cι _-4 -

Alkanoyl, C _1-4 alkoxycarbonyl, aminocarbonyl and aryl,

independently one or two radicals selected from the

Group comprising:

Hydrogen, F, Cl, Br,

OH, OR ⁴ , OCOR ⁴ , OCONHR ⁴ ,

COR ⁴ ,

CN, COOH, COOR ⁴ , CONH ₂ , CONHR ⁴ , CONRV, CONHOH,

CONHOR ⁴ , SR ⁴ , SOR ⁴ , SO ₂ R ⁴ , SO ₂ NH ₂ , SO ₂ NHR ⁴ , SO ₂ NRV ^' ,

N0 ₂ , NH ₂ , NHR ⁴ , NRV,

NHSO2R ^4, NR ⁴ S0 ₂ R ^{4 ',} NHSO2R ^6, RrS0 ₂ R ^6, NHCOR ⁴ , NHCOOR ⁴ , NHCONHR ⁴ and R ⁴ , where the radicals R ⁴ , R ⁴ and R ⁶ are selected independently of one another according to the meanings given below,

A is a group selected from the group comprising CMO

Alkanediyl, C ₂ -ιo alkenediyl, C _2- ιo alkynediyl and (co- ₃ alkanediyl-C _3-7 _{cycloalkanediyl-co-3} alkanediyl), wherein in a five-membered cycloalkyl ring one ring member ring N or ring 0 can be and in a six- or seven-membered

Cycloalkyl ring may be one or two ring members each ring N and / or ring O atoms, wherein the ring N atoms may optionally be substituted by C-ι _-3 alkyl or C _1-3 alkanoyl, wherein in the above-mentioned aliphatic chains one C atom from O, NH, N-alkyl or N-Cι _-3 _-3 Cι alkanoyl may be exchanged, and wherein alkyl or cycloalkyl optionally substituted with one radical selected from the group comprising = 0, OH, Od ₃ alkyl, NH _2, NH-Cι _-3 alkyl, NH-C _1-3 -alkanoyl, N (C _1-3 alkyl) ₂ and N (Cι _-3 - AlkyiχC-ι _3- alkanoyl), may be substituted,

8 is a radical selected from the group comprising COOH, COOR ⁵ ,

CONH ₂ , CONHNH ₂ , CONHR ⁵ , CONR ⁵ R ^{5 '} , CONHOH, CONHOR ⁵ and tetrazolyl, each bonded to a C atom of group A, wherein the radicals R ⁵ and R ^{5 are} independently selected according to the meanings given below .

V is a group selected from the group comprising O, NH,

NR ^ NCOR ⁴ , NSO ₂ R ⁴ and NSO ₂ R ⁶ , where R ⁴ and R ^{6 have} the meanings given below, wherein in the above radicals the radicals R ⁴ , R ^{4 '} , R ⁵ , R ^5' and R ^{6 have} the following meanings; in it are:

R ⁴ and R ^{4 '} are each independently a radical selected from among

Group comprising CF ₃ , C ₂ F ₅ , Cι _-4 alkyl, C _2- alkenyl, C _2-3 alkynyl and

(C _0-3 alkanediyl-C _3-7 cycloalkyl), wherein in a five-membered cycloalkyl ring, a ring member may be ring N or ring O and in a six- or seven-membered

Cycloalkyl ring, one or two ring members can be ring N and / or ring O atoms, respectively, where the ring N atoms with Cι _-3 alkyl or Cι _-3- alkanoyl may be optionally substituted,

R ⁵ and R ⁵ 'are each independently a radical selected from among

Group comprising Cι- ₆ alkyl, C _2-6 alkenyl, C _2-6 alkynyl, wherein a carbon atom to O, S, SO, S0 _2, NH, NC _1-3 alkyl or NC _{1 3} alkanoyl may be exchanged, further (C _0-3 alkanediyl-C _3-7 cycloalkyl), wherein in a five-membered cycloalkyl ring may be a ring member ring N or ring-O and in a six- or seven-membered cycloalkyl ring two ring members may each be ring N and / or ring O atoms, whereby the ring N atoms _-3 alkanoyl may be optionally substituted with Cι _-3 alkyl or Cι, and also (co ₃ -alkanediyl Aryl) and (C _0-3 alkanediyl)

Heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, all of the aforementioned alkyl and cycloalkyl radicals having up to two radicals selected from the group comprising CF ₃ , C ₂ F ₅ ,

OH, 0-Cι _-3 alkyl, NH _2, NH-C _1-3 alkyl, NH-Cι _-3 alkanoyl, N (C _1-3 - alkyl) _2, N (C _1-3 alkyl) (C ₃ alkanoyl), COOH, CONH ₂ and COO-Cι _-3 - alkyl, and all of the aforementioned aryl and heteroaryl groups with bis to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C ₂ H ₅ , OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , and / or can also carry a fused methanediylbisoxy or ethane-1, 2-diylbisoxygruppe, or R ⁵ and R ⁵ together with the amide N atom of B is a five- to seven-membered , saturated or unsaturated heterocyclic ring which may further contain N or O or S atom and which may be substituted with Cι _-4 alkyl, (co ₂ alkanediyl-Cι _-4 -alkoxy) C-ι _-4 -alkoxycarbonyl, aminocarbonyl or aryl,

R ^{6 is} a radical selected from the group comprising (C _0-3 -alkanediyl)

Aryl) and (co-3-alkanediyl-heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, and wherein the

Aryl and heteroaryl groups may be substituted with up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C 2 H 5, OH, OCH ₃ , OC ₂ H 5, NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , and / or can also carry a fused methanediylbisoxy or ethane-1, 2-diylbisoxy group,

R ⁷ and R ^{7 '} independently of one another R ⁴ or R ⁶ , for the preparation of a medicament for the treatment of a disease according to any one of claims 1-5.

7. Use of a benzimidazole according to claim 6, characterized in that

F, Cl, Br, C (NH) NH ₂ , C (NH) NHR ⁴ , C (NH) NR ⁴ R ⁴ ', C (NR ⁴ ) NH ₂ , C (NR ⁴ ) NHR ⁴ ', C (NR ⁴ ) NR R ⁴ ' .

OH, OR ⁴ , OCOR ⁴ , OCONHR ⁴ , COR ⁴ , C (NOH) R ⁴ , CN, COOH, COOR ⁴ , CONH ₂ , CONRV ', CONHR ⁴ , CONHOH,

SR ^4, SOR ^4, Soar ^4, SO2NH2, S0 ₂ NHR ^4, S0 ₂ NR ⁴ R ^{4 ',} N0 _2l NH _2, NHR ^4, ^NRV', NHCONHR ⁴ and

R ⁴ , wherein the radicals R ⁴ and R ⁴ indicated below

Meanings are independently selected and wherein two substituents on R ¹ can be linked together so that they together form a Methandiylbisoxy-, ethane-1, 2-diylbisoxy, propane-1, 3-diyl or butane-1, 4-diyl group form if they are orthostable to each other,

a mono- or bicyclic Cβ-io-aryl group or a mono- or bicyclic 5-10-membered heteroaryl group with 1-2

Heteroatoms selected from the group consisting of N, S or O, wherein said aryl or heteroaryl group may be substituted with up to three of the following substituents independently of one another:

F, Cl, Br,

XOH, XOR ⁴ , XOCOR ⁴ , XOCONHR ⁴ , XOCOOR ⁴ , XCOR ⁴ , XC (NOH) R ⁴ , XC (NOR ⁴ ), XC (NO (COR ⁴ )) R ⁴ ,

XCOOH, XCOOR ⁴ , XCONH 2, XCONHR ⁴ , XCONR ⁴ R ⁴ , XCONHOH,

XCONHOR ⁴ , XCOSR ⁴ ,

XSR ^4, XSOR ^4, XSO2R ^4, SO2NH2, S02NHR ^4, S0 ₂ NR ⁴ R ^4,

N0 _2, XNHR ^4, XNR ⁴ R ^4, XNHS0 ₂ R ^4, XN (S0 ₂ R ⁴⁾ S0 ₂ R ^4, XNR ⁴ S0 ₂ R ^4, R ^4, where two substituents on R ^2, if they are ortho to one another , so they can be linked together Methanediylbisoxy, ethane-1,2-diylbisoxy, propane-1,3-diyl, butane-1, 4-diyl,

R ^{3 is} a radical selected from the group comprising hydrogen, F, Cl, Br, CH ₃ , C ₂ H ₅ , CF ₃ , C ₂ F ₅ , OH, OR ⁴ , NHSO ₂ R ⁶ and NHCOR ⁴ , wherein R ⁴ and R ^{6 have} the meanings given below,

A Ci.-io-alkanediyl, C ₂ -ιo-alkenediyl, C- ₂ -ιo-alindiyl, (C ₀ -5-alkanediyl-C _3-7 - cycloalkanediyl-co-5-alkanediyl), where in a 5- be membered cycloalkyl ring, a ring member may be an N or an O and may be in a 6- or 7-membered cycloalkyl ring one or two ring members N and / or 0, wherein ring nitrogens optionally with C-ι _-3 alkyl or C ₃ alkanoyl can be substituted, in the above-mentioned aliphatic chains one carbon atom or two carbon atoms for O, NH, NCι _-3 - ₃ alkanoyl can be replaced alkyl, NC-ι-,

B is a radical selected from the group comprising COOH, COOR ⁵ ,

CONH ₂ , CONHR ⁵ and CONR ^ ^' , each attached to a C atom of group A, wherein the radicals R ⁵ and R ^5' are selected independently of one another according to the meanings given below,

O

R ⁴ and R ^{4 are the} same as stated above, R ⁵ and R ⁵ are each independently a radical selected from among

Group comprising C _1-6 alkyl, C _2- 6 alkenyl, C ₂ -6-alkynyl, where one carbon atom to O, S, SO, S0 _2, NH, NC _1-3 alkyl or NC _{1 -3} alkanoyl may be replaced, further (co-3-alkanediyl-C3 cycloalkyl), wherein ring 0, in a five-membered cycloalkyl ring one ring member ring N or and in one six- or seven-membered cycloalkyl ring or two ring members each of ring N and / or ring O atoms may be, where the ring N atoms _-3 alkanoyl may be optionally substituted with Cι _-3 alkyl or Cι, and also (co-3-alkanediyl -Phenyl) and

(Co-3-alkanediyl-heteroaryl) wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, with all of the aforementioned alkyl and cycloalkyl radicals having one radical selected from the group comprising _CF3, C2F5, OH, OCi.

3-alkyl, NH _2> NH-Cι _-3 alkyl, NH-Cι _-3 alkanoyl, N (C _1-3 alkyl) _2, N (C _1-3 - alkyl) (C _1-3 alkanoyl ), COOH, CONH ₂ and COO-C _1-3 alkyl, and all of the aforementioned phenyl and heteroaryl groups having up to two radicals selected from the group comprising F, Cl, Br, CH ₃ , C 2 H 5, OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , and / or may also carry a fused methanediylbisoxy or ethane-1,2-diylbisoxy group or R ⁵ and R ⁵ together with the amide N atom of β form a five- to seven-membered, saturated or unsaturated heterocyclic ring containing another N- or O- or S-

May contain atom and may be substituted by the C _1-4 alkyl, (Co-2-alkanediyl Cι _-4 alkoxy), Cι- alkoxycarbonyl, aminocarbonyl or phenyl,

R ^{6 is} a phenyl or heteroaryl group, wherein the heteroaryl group is five- or six-membered and contains one or two heteroatoms selected from the group comprising N, S and O, and wherein the phenyl and heteroaryl groups substituted with up to two radicals selected from the group comprising F, Cl, Br, CH _3> C ₂ H _5l OH, OCH ₃ , OC ₂ H ₅ , NO ₂ , N (CH ₃ ) ₂ , CF ₃ , C ₂ F ₅ and S0 ₂ NH ₂ , or can also carry a fused Methandiylbisoxy- or ethane-1, 2-diylbisoxygruppe, means.

8. Use of a benzimidazole according to claim 6 or 7, characterized in that R ^{3 is} hydrogen and Y- A d ₆ -alkyleneoxy.