BG63361B1 - New substituted imidazoles - Google Patents

Abstract

The invention relates to new 1,4,5-substituted imidazoles and compositions to be used as cytokin inhibitors. 25 claims

Description

(54) NEW REPLACED IMIDAZOLES

Technical field

The invention relates to a new group of imidazoles, to a method for their preparation and to their use for the treatment of cytokine mediator diseases, as well as for pharmaceutical compositions for use therein.

BACKGROUND OF THE INVENTION

Interleukin-1 (IL-1) and tumor necrosis factor (TNF) are biological substances produced by various cells such as monocytes or macrophages. IL-1 is known to mediate various biological activities considered important in immunoregulation and other physiological conditions such as inflammation [see, e.g. Dinarello et al., Rev. Infect. Disease, 6, 51 (1984)]. The many known biological activities of IL-1 include activation of T-helper cells, induction of inflammation, stimulation of prostaglandin or collagenase production, neutrophil chemotaxis, induction of acute-phase proteins, and suppression of plasma iron levels.

There are many conditions in which excessive or unregulated production of IL-1 is involved in exacerbating and / or causing the disease. These include rheumatoid arthritis, osteoarthritis, endotoxemia and / or toxic shock syndrome, other acute or chronic inflammatory conditions such as endotoxin-induced inflammatory response or bowel inflammation; tuberculosis, atherosclerosis, muscle degeneration, cachexia, psoriasis arthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis and acute synovitis. New data also link the effects of IL-I with diabetes and pancreatic β-cells.

Dinarello, J. Clinical Immunology, 5 (5), 287-297 (1985) provides an overview of the biological activities inherent in IL-1. It should be noted that some of these effects have been described by other authors as indirect actions of IL-1.

Excessive or unregulated production of THF is involved in the mediation or exacerbation of many diseases, including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions; septicemia, septic shock, endotoxic shock, Gram-negative sepsis, toxic shock syndrome, respiratory distress syndrome in adults, cerebral malaria, chronic pulmonary inflammation, silicosis, pulmonary sarcoisosis, bone resorption transplantation, bone disease transplantation, allograft, fever and myalgia due to infection such as influenza, cachexia due to infection malignancy, cachexia due to acquired immune deficiency syndrome (AIDS), AIDS (AIDS) ) associated with spin complex (ARC), keloid formation, cicatrix tissue formation, Crohn's disease, ulcerative colitis, or pyresis.

AIDS is the result of human immunodeficiency virus (HIV) infection of Tymphocytes.

At least three types of HIV strains have been identified, ie. HIV-1, HIV-2 and HIV-3. As a result of HIV infection, immunity with mediator T cells is impaired and infected individuals show severe conditionally pathogenic infections and / or unusual neoplasms. The entry of HIV into T lymphocytes requires activation of T lymphocytes. Other viruses such as HIV-1, HIV-2 infect T lymphocytes after T cell activation and such viral protein expression and / or replication is mediated or maintained by such T cell activation. Once activated T-lymphocyte is infected with HIV, T-lymphocyte can continue to be maintained in the activated state to allow expression of the HIV gene and / or HIV replication.

Monokines, in particular TNF, are involved in the expression of HIV protein and / or replication of viruses with mediated activated T cells, playing a role in maintaining the activation of T lymphocytes. Therefore, interference with the activity of monokines, for example, by inhibiting the production of monokines, in particular TNF, in HIV-infected individuals helps to limit the maintenance of T-cell activation, thereby reducing the progression of HIV infection in uninfected cells, leading to a delay or elimination of the progression of immune dysfunction caused by HIV infection. Monocytes, macrophages and related cells, such as Kupffer and glial cells, are also involved in the maintenance of HIV infection. These cells, like T cells, are targets for virus replication and the level of viral replication depends on the activated state of the cells.

Monokines, such as TNF, have been shown to activate HIV replication in monocytes and / or macrophages [See Poli et al., Proc. Natl. Acad, Sci., 87: 782-784 (1990)], therefore, inhibiting the production of monokines or activity helps to limit the progression of HIV, as stated in the Cells above.

TNF implements various functions with other viral infections such as cytomegaly virus (CMV), influenza virus and herpes virus for the same reasons as noted.

Interleukin-8 (IL-8) is a chemotaxis factor identified and characterized for the first time in 1987. IL-8 is produced by several cell types, including mononuclear cells, fibroblasts, endothelial cells and keratinocytes.

Its production by endothelial cells is induced by IL-1, TNF or lipopolysaccharide (LPS). Human IL-8 has been found to act on the neutrophils of mouse, guinea pig, rat and rabbit. Many different names are given to IL-8, such as neutrophil attracting protein (NAP-1), derived from monocyte neutrophil chemotaxis factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte chemotaxis factor.

IL-8 stimulates many functions in vitro. It has been shown to have chemoattractive properties for neutrophils, T-lymphocytes and basophils. In addition, it induces the release of histamine from basophils by normal and atopic individuals, as well as the release of lysosomal enzyme and respiratory impulse from neutrophils. IL-8 is known to increase the expression surface of Mas-1 (CDIb / CD18) on neutrophils without new protein synthesis, which may promote neutrophil adhesion to vascular endothelial cells. Many diseases are characterized by massive neutrophil infiltration.

Conditions associated with increased production of IL-8 (which is responsible for the chemotaxis of neutrophil cells at the site of inflammation) can be ameliorated by compounds that suppress IL-8 production.

IL-1 and TNF refer to different cells and tissues, and these cytokines, as well as others derived from leukocyte cytokines, are important and critical mediators of inflammation in a wide variety of diseases and conditions. Inhibition of these cytokines is useful in controlling, reducing, and alleviating many of the disease states.

In this area there is always a need for treatment and for cytokine suppressants, anti-inflammatory drugs, i. compounds that are capable of inhibiting cytokines such as IL-1, IL-6, IL-8 and TNF.

SUMMARY OF THE INVENTION

The present invention relates to novel compounds of formula I, to pharmaceutical compositions containing compounds of formula I, and a pharmaceutically acceptable diluent or nose;

The present invention also relates to a method for the treatment of mammalian diseases with a mediator CSBP / RK / p38 kinase, which comprises administering to the mammals in need of treatment an effective amount of a compound of formula I.

The present invention relates to a method for inhibiting cytokines and treating mammalian diseases with a cytokine mediator, which comprises administering to the mammals in need of treatment an effective amount of a compound of formula I.

The present invention relates in particular to a method of inhibiting the production of IL-1 in mammals, which comprises administering an effective amount of a compound of formula I.

The present invention relates in particular to a method of inhibiting the production of IL-8 in mammals, which comprises administering an effective amount of a compound of formula I.

The present invention relates in particular to a method of inhibiting the production of TNF in mammals, which comprises administering an effective amount of a compound of formula I. 5

The present invention accordingly relates to compounds of formula I

wherein R 1 represents a 4-pyrimidinyl ring substituted with C ₁₄ alkoxy and optionally optionally substituted independently with C _1-4 al-15 kilyl, halogen, hydroxy, Q ₄ alkoxy, C ₁₄ alkylthio, C ₁₄ alkylsulfinyl, CH. OR ₁₂ , amino, mono and di-C ₁₆ alkyl substituted amino, N (R ₁₀ ) C (O) R _c or N-heterocyclic ring having a ring having from 5 to 7 atoms and optionally containing 20 additional heteroatom - oxygen , sulfur or NR ₁₅ ;

R ₄ stands for phenyl, naphth-1-yl or naphth-2-yl, or heteroaryl, which is optionally substituted by one or two substituents, each of which 25 is independently selected, and which is a substituent on 4-phenyl, 4-naphth-1 -yl, 5-naphth-2-yl or 6 naphth-2-yl is halogen, cyano, nitro, C (Z) NR ₇ R ₁₇ , C (Z) OR ₁₆ , (CR ₁₀ R ₂₀ ) COR ₁₂ , SR ₅ , SOR ₅ , OR ₁₂ , substituted with halogen C ₁₄ alkyl, C ₁₄ alkyl, ZC (Z) R ₁₂ , 30 NR ₁₀ C (Z) R ₁₆ or (CR ₁₀ R ₂₀ ) _v NRi ₀ R _20, and which is a substituent for the other place of substitution is halogen, cyano, C (Z) NR ₁₃ R ₁₄ , C (Z) OR ₃ , (CR ₁₀ R ₂₀ ) _m. COR ₃ , S (O) _m R ₃ , OR ₃ substituted with halogen C ₁₄ alkyl, C 1-4 alkyl (CR ₁₀ R ₂₀ ) _m ., NR ₁₀ (Z) R ₃ , NR, _O S (O) _m. R ₈ , 35 NR, _O S (O) _m . NR.R ,,, ZC (Z) Rj, or (CR ₁₀ R _{20) m,} NR _I3 R _14;

n is 1 to 10;

m is 0, 1 or 2;

m 'is a number 1 or 2;

m ”is 0 from 1 to 5; 40 in e is 0 or 1 or 2;

R ₂ is optionally substituted heterocycle or optionally substituted heterocyclic C 1. _{1 #} alkyl residue;

Z is oxygen or sulfur; 45

R _c is hydrogen, C ₁₆ alkyl, C _3-7 cycloalkyl, aryl, aryl C ₁₄ alkyl, heteroaryl, heteroaryl C ₁₄ alkyl, heterocycle or heterocyclic C _{| 4} alkyl;

R ₃ is a heterocycle, heterocyclic C ₁₁₀ 50 allyl or R _g ;

R ₃ is hydrogen, C ₁₄ alkyl, C ₂₄ alkenyl,

C _2-4 alkynyl or NR ₇ R ₁₇ , except when SR ₃ is SNR ₇ R ₁₇ and SOR ₅ is SOH;

R 1 and R ₁₇ are independently hydrogen or C ₁₄ alkyl or R ₇ and R ₁₇ together with the nitrogen to which they are attached form a heterocyclic ring of 5 to 7 atoms, which ring optionally contains an additional heteroatom, oxygen, sulfur or NR ₁₅ ;

R _g is C ₁₁₀ alkyl substituted with halogen C ₁₁₀ alkyl, C ₂₁₀ alkenyl, C ₂₁₀ alkynyl, C ₃ . ₇ cycloalkyl, C ₅ . ₇ cycloalkenyl, aryl, aryl C ₁₁₀ alkyl, heteroaryl, heteroaryl C 1-6 alkyl, (CR ₁₀ R ₂₀ OR,), (CR ₁₀ R ₂₀ ) _n S (O) _m R ₁₈ , (CR ₁₀ R ₂₀ ) NHS ( O) ₂ R, _g, (CR _I0 R _{20) n} NR _l3 R ₁₄ as residues: aryl, arylalkyl, heteroaryl, heteroarylalkyl can optionally be substituted;

R 1 is hydrogen, C (Z) R _H or optionally substituted Ό,., Θ alkyl, S (O) ₂ R _1g , optionally substituted aryl or optionally substituted aryl C ₁₄ alkyl;

R ₁₀ and R _{20 are} each independently hydrogen or C ₁₄ alkyl;

R _H is hydrogen, C 1-4. ₁₀ alkyl, C ₃ . ₇ cycloalkyl, heterocyclyl, heterocyclyl _C1-10 alkyl, aryl, aryl C ,. ₁₀ alkyl, heteroaryl or heteroaryl C ₁₀ alkyl;

R ₁₂ is hydrogen or R ₁₆ ;

R ₁₃ and R ₁₄ are all independently of one another hydrogen or optionally substituted C _1-4 alkyl, optionally substituted aryl, or optionally substituted aryl alkyl, or together with the nitrogen atom to which they are attached form a heterocyclic a ring of 5 to 7 atoms, which ring optionally contains additional heteroatom oxygen, sulfur or NR ₉ ;

R ₁₅ is R ₁₀ or C (Z) -C ₁₄ alkyl;

R ₁₆ is C _b4 alkyl, halosubstituted C, ₄ alkyl or C ₃ _ ₇ cycloalkyl;

R _lg is C _{11 ()} alkyl, _C3-7 cycloalkyl, heterocyclyl, aryl, arylalkyl, heterocyclyl, heterocyclyl ,. _{| 0} alkyl, heteroaryl or heteroaryl C _{1 | 0} alkyl;

or pharmaceutically acceptable salts thereof.

The novel compounds of formula I, except in humans, may also be used in the veterinary treatment of mammals; that need to inhibit cytokine inhibition or production. In particular, diseases with a cytokine mediator that will be treated therapeutically or prophylactically in animals include disease states such as those indicated in the "Treatment methods" section, in particular viral infections.

Examples of such viruses include, but are not limited to, lentiviral infections, such as infectious anemia virus in horses, goat arthritis virus, visna virus, or maedi virus, or retroviral infections such as, but not limited to, virus, immune cat failure (FIV), bovine immunodeficiency virus, or dog immune deficiency virus or other retroviral infections.

In formula IR _I (the 4-pyrimidinyl ring) is substituted at least once with a C _1-4 alkoxy group.

Preferred is C, _ ₄ alkoxy group such as n-butyl, isopropoxy, ethoxy or methoxy. The preferred site of the substituent in the 4-pyrimidinyl ring is R ₍ is the II site as 2-methoxy-4-pyrimidinyl).

Suitable other substituents for the R, heteroaryl ring are C _1-4 alkyl, halogen, OH, C ₁₄ alkoxy, C ₁₄ alkylthio, C ₁₄ alkylsulfinyl, CH ₂ OR ₁₂ , amino, mono- and C ₁ -C ₁₆ alkyl substituted amino, N ( R ₁₀ ) a C (O) R _c or N-heterocyclyl ring having a ring having 5 to 7 atoms and optionally containing an additional heteroatom - oxygen, sulfur or NR ₁₅ . The alkyl group in the mono- and C ₁ -C ₁₆ alkyl substituted moiety may be halogen substituted, such as trifluoro-, for example trifluoromethyl or trifluoroethyl.

When, in one case, the substituent of R ₁ is N (R ₁₀ ) C (O) R _c , where R _c is hydrogen, C ₁₆ alkyl, C ₃ . ₇ cycloalkyl, aryl, aryl C _1-4 alkyl, heteroaryl C _1-4 alkyl, heterocyclyl or heterocyclyl C _b4 alkyl C, _ ₄ alkyl, R _c is preferably C ₆ alkyl; R ₁₀ is preferably hydrogen.

It should be appreciated that R _c , in particular the C _1-6 alkyl group, may optionally be substituted, preferably from one to three times, preferably by halogen such as fluorine such as trifluoromethyl or trifluoroethyl.

Suitably, R ₄ is phenyl, naphth-1-yl or naphth-2-yl, or heteroaryl, which is optionally substituted by one or two substituents. Preferably, when R ₄ is a phenyl or naphthyl ring. Suitable substituents for R ₄ when 4-phenyl, 4-naphth-1-yl, 5-naphth-2-yl or 6-naphth-2-yl are one or two substituents, each of which independently of one another is halogen, -SR ₅ , -SOR, -OR ₁₂ , CF ₃ or - (CR ₁₀ R ₂₀ ) NR, _O R _W and for the other substitution position in these rings the preferred substituent is halogen, -S (O) _m R ₃ , -OR ₃ , CF ₃ , - (CR, _O R ₂₀ ) _m 'NR ₁₃ R ₁₄ , -NR ₁₀ C (Z) R ₃ and -NR ₁₀ S (O) _m , R ₈ .

Preferred substituents for the 4-position in the phenyl naphth-1-yl ring and the 5-position in the naphthyl-2-yl ring are halogen, especially fluorine and chlorine, and -SR ₅ and -SOR 2, such as R, is preferably C. ₂ alkyl, in particular methyl; of which fluorine and chlorine are more preferred and the fluorine substituent is particularly preferred. Preferred substituents for the 3-position in the phenyl and naphth-1-yl rings are halogen, in particular fluorine or chlorine; -OR ₃ , in particular C ₁₄ alkoxy; CF _3, NR ₁₀ R, ₀ as amino; -NR ₁₀ C (Z) R ₃ , in particular NHCOICl (alkyl); -NR | _O S (O) _m , R _s , in particular -NHSO, (C ₁₁₀ alkyl) and -SR ₃ and -SOR ₃ , wherein R ₃ is preferably C 1-6 alkyl, more preferably methyl. When the phenyl ring is disubstituted, it is preferably two independent halogen substituents, such as fluorine and chlorine, preferably dichloro and more preferably at the 3,4-position. Both -OR ₃ and -ZC (Z) R ₃ substituents are also preferred for the 3 position, and R ₃ may also be hydrogen.

Preferably R ₄ is an unsubstituted or substituted phenyl radical. More preferably, R ₄ is phenyl or phenyl substituted in 4 position with fluorine and / or substituted in 3 place by fluorine, chlorine, C _1-4 alkoxy, methanesulfonamido or acetamido or R ₄ is phenyl disubstituted in 3,4 position independently of chlorine or fluorine, preferably chlorine. It is particularly preferred that R ₄ be 4-fluorophenyl.

In formula I, Z is oxygen or sulfur, preferably oxygen.

It is recommended that R ₂ is optionally substituted heterocycle or heterocyclic Cj_ | ₀ alkyl radical.

When R ₂ is optionally substituted heterocycle, the ring is preferably in the group of morpholino, pyrrolidinyl or piperidinyl. When the ring is optionally substituted, the substituent can be directly attached to the free nitrogen as well as a piperidinyl group either in the pyrrole ring or in the ring itself. Preferably the ring 5 is piperidine or pyrrole, more preferably piperidine. The heterocyclic ring may optionally be substituted one or four times independently by halogen; C ₁ . ₄ alkyl, aryl such as phenyl; arylalkyl such as benzyl, where the aryl or arylalkyl radical itself may be optionally substituted (as defined below); C (O) OR 11 such as C (O) C ₁₄ alkyl or C (O) OH groups; C (O) H; C (O) C _1-4 alkyl, hydroxy C, ₄ alkyl, _{alkoxy, S (O) m C l} _ ₄ alkyl (wherein m is 0,1 or 2), NR ₁₀ R ₂₀ (wherein R ₁₀ and R ₂₀ are independently of one another hydrogen or C ₁₄ alkyl).

Preferably, if the ring is piperidine, it is attached to the imidazole in 4 position and the substituents are directly attached to the nitrogen atom, for example 1-formyl-4-piperidine, 1-benzyl-4-piperidine, 1-methyl-4-piperidine, 1-ethoxycarbonyl- 4-piperidine. If the ring is substituted with an alkyl group and is attached at the 4-position, it is preferable that the substituent be in the 2-position or 6-position or both, such as 2,2,6,6-tetramethyl-4-piperidine. Similarly, if the ring is pyrrole, it is attached to the imidazole in 3 position and all substituents are directly attached to the nitrogen atom.

When R ₂ is optionally substituted by a heterocyclic C _1-10 alkyl group, the ring is preferably morpholine, pyrrolidinyl or piperidinyl. Preferably the alkyl group has 1 to 4 carbon atoms, more preferably 3 or 4 and most preferably 3, as in the propyl group. Preferably, heterocycloalkyl groups include, but are not limited to, morpholinoethyl, morpholinopropyl, pyrrolidinylpropyl and piperidinylpropyl. The heterocycle ring herein is also optionally substituted in a similar manner to that indicated above for the direct binding of the heterocycle.

In any case, where there is an alkenyl or alkynyl radical as a substituent, the unsaturated bond, for example the vinylene or acetylene bond, is preferably not directly bonded to nitrogen, oxygen or sulfur, for example in OR ₃ or in some R ₂ residues.

As used herein, the term "optionally substituted", unless otherwise indicated, means that halogen groups such as fluorine, chlorine, bromine or iodine; hydroxy substituted C _10-10 alkyl; C ₁₀ alkoxy such as methoxy or ethoxy; S (O) alkyl, wherein m is 0,1 or 2 as methylthio, m ^{7 is} methylsulfinyl or methylsulfonyl; an amino, mono- and disubstituted amino such as the NR ₇ R ₁₇ group; or as R ₇ R ₁₇ may, together with the nitrogen to which they are attached, cyclize to form a 5- to 7-atom ring which optionally includes an additional heteroatom selected from O / N / S; With |. | _An alkyl, cycloalkyl or cycloalkylalkyl group such as methyl, ethyl, propyl, isopropyl, tert-butyl, etc. or cyclopropylmethyl; halogen substituted C _0-10 alkyl such as CFC, F ₂ H or CF ₃ ; halosubstituted C, _ ₁₀ alkoxy such as OCF ₂ CF ₂ H; optionally substituted aryl such as phenyl or optionally substituted arylalkyl such as benzyl or phenethyl, the aryl substituent may also be substituted once or twice with halogen; hydroxy; hydroxy substituted alkyl; With ₁₁₀ alkoxy; S (0) _da alkyl; an amino, mono- or disubstituted amino such as the NR ₇ R ₁₇ group; alkyl, or CF _3.

In a preferred embodiment of the compound of formula I, R 1 is 2-alkoxy-4-pyrimidinyl; R ₂ is morpholinylpropyl, piperidinyl, N-benzyl4-piperidinyl or N-methyl-4-piperidinyl and R ₄ is phenyl or mono- or disubstituted phenyl with fluorine, chlorine, C _1-4 alkoxy, -S (O) _f 1 alkyl, methanesulfonamido or acetamido.

Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include alkali salts of inorganic and organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic, acetic, malic, tartaric, citric, mint , succinic, fumaric, maleic, benzoic, salicylic, phenylacetic and almond acid. In addition, pharmaceutically acceptable salts of compounds of formula I may also be formed with a pharmaceutically acceptable cation, for example, if the substituent group contains a carboxy residue. Suitable pharmaceutically acceptable cations are well known in the art and include alkali, alkaline earth, ammonium and quaternary ammonium cations.

The following terms, as used herein, refer to:

- "halogen" or "halogens", includes halogens chlorine, fluorine, bromine and iodine.

- "C _1-10 alkyl" or "alkyl" both straight and branched chain radicals of 1 to 10 carbon atoms, unless the chain length is otherwise limited, include (but are not limited to) methyl, ethyl, n. propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl and the like.

- The term "cycloalkyl" is used herein to refer to cyclic radicals, preferably with 3 to 8 carbon atoms, including, but not limited to, cyclopropyl, cyclopentyl, cyclohexyl and the like.

- The term "cycloalkenyl" is used herein to refer to cyclic radicals, preferably with 5 to 8 carbon atoms having at least one bond, including (but not limited to) cyclopentyl, cyclohexyl and the like.

- The term "alkenyl" used herein in all cases means a straight or branched chain 2-10 carbon atom, unless the chain length is broken includes (but not limited to) ethenyl, 1-propenyl, 2- propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.

- "aryl" means phenyl or naphthyl.

- "heteroaryl" (as such or in combination as "heteroaryloxy" or "heteroarylalkyl") means an aromatic ring system of 5-10 atoms in which one or more rings contain one or more heteroatoms Ν, O or S as, but not limited to them, pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrlimidine, oxazole, thiazole, thiadiazole, triazole, imidazole or benzimidazole.

- "heterocyclic" moiety (as such or in combination as "heterocyclylalkyl) means a saturated or partially unsaturated 410 atomic ring system in which one or more rings contain one or more heteroatoms - N, 0, or S as (but not limited to) pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran or imidazolidine.

- The term "aralkyl" or "heteroarylalkyl" or heterocycloalkyl "as used herein refers to C ₁₄ alkyl as defined above, linked to an aryl, heteroaryl or heterocyclic moiety as defined, unless otherwise stated:

- "sulfinyl" refers to the S (0) of the corresponding sulfide, the designation "thio" refers to the sulfide and the designation "sulfonyl" refers to the fully oxidized S (0) ₂ residue.

- "aroyl" refers to C (O) Ar, where Ar denotes phenyl, naphthyl or arylalkyl as defined above, including but not limited to benzyl and phenethyl.

- "alkanoyl" refers to C (O) C _{| J is} alkyl, where alkyl is as defined above.

According to the invention, the 4-pyrimidinyl radical R or R ₂ has the formula

The compounds of the invention may contain one or two asymmetric carbon atoms and may exist in racemic or optically active forms. All of these compounds are within the scope of the present invention.

Examples of compounds of formula I are:

- (4-piperidinyl) -4- (4-fluorophenyl) -5 (2-iso-propoxy-2-pyrimidinyl) imidazole

- (4-piperidinyl) -4- (4-fluorophenyl) -5 (2-methoxy-34-pyrimidinyl) imidazole

5- (2-hydroxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

5- (2-methylthio-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

5- (2-methylthio-4-pyrimidinyl) -4- (4-fluorophenyl) -1 - [(1-methyl-4-piperidinyl] imidazole

5- (2-ethoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

- (1-ethylcarboxylpiperidin-4-yl) -3- (4-thiomethylphenyl) -5- [2- (thiomethyl) -pyrimidin-4-yl] imidazole

- (1-ethylcarbonylpiperidin-4-yl) -4- (4methylsulfinylphenyl) -5- [2-methylsulfinylpyrimidin-4-yl] imidazole.

A preferred group of compounds of formula I have the following structure:

wherein R, is pyrimidinyl substituted with a C, _ ₄ alkoxy, and optionally further substituted one or more times independently with C ₄ alkyl, halogen, hydroxy, C _t _ ₄ alkoxy, C _1-4 alkylthio, C _1-4 alkylsulfinyl, CH ₂ OR ₁₂ , an amino, mono- or di-C _1-4 alkyl substituted amino, N (R ₁₀ ) C (O) R _c or N-heterocyclic ring of 5 to 7 atoms and optionally containing an additional heteroatom - oxygen, sulfur or NR ₁₅ ;

R ₂ is an optionally substituted heterocyclyl or optionally substituted heterocyclyl C _{1 | 0} alkyl radical;

R ₄ is phenyl which is optionally substituted by halogen;

R ₁₀ is independently selected from hydrogen or _b4 alkyl;

R _c is hydrogen, C ₁₆ alkyl, C _3-7 cycloalkyl, aryl, aryl C ₁₄ alkyl, heteroaryl, heteroaryl C alkyl, heterocyclyl or heterocyclyl C ₁₄ alkyl C ₁₄ alkyl, all of which may be optionally substituted;

R ₁₂ is hydrogen or R ₁₆ ;

R _lt is C 1-6 alkyl, C 1-4 alkyl substituted with halogen or C _3-7 cycloalkyl;

R ₁₅ is hydrogen, C ₁₄ alkyl or C (Z) -C ₁₄ alkyl;

Z is oxygen or sulfur or their pharmaceutically acceptable salts.

Preferably R ₂ is piperidine, 1-formyl-4-piperidine, 1-benzyl-4-piperidine, 1-methyl-4-piperidine, 1-ethoxycarbonyl-4-piperidine, 2,2,6,6-tetramethyl-4 -piperidine, morpholinoethyl, morpholinopropyl, pyrrolidinylpropyl or piperidinylpropyl.

The compounds of formula I can be prepared using syntheses, some of which are illustrated in the following schemes applicable to the preparation of compounds of formula I, with different R 1, R ₂ and R ₄ substituent groups which are reacted using optionally substituents that are suitably protected to be compatible with the reaction described below. Subsequent removal of the protecting group, in these cases, provides compounds with generally described properties. Once an imidazole nucleus is formed, other compounds of formula I can be prepared using standard functional group conversion techniques well known to those skilled in the art.

For example, -C (O) NR ₁₃ R ₁₄ can be obtained from CO ₂ CH ₃ by heating, in the presence or absence of a catalyst, of metallic cyanide, e.g. NaCN and NHR ₁₃ R ₁₄ in CH ₃ OH; -OC (O) R ₃ is obtained from OH for example with CIC (O) R in pyridine; -NR ₁₀ -C (S) NR ₁₃ R ₁₄ is prepared from -NHR ₁₀ with alkylisocyanate or thiocyanic acid;

NR ₆ C (O) OR ₆ is obtained from -NHR ₆ c-alkylchloroformate; -NR ₁₀ C (O) NR ₁₃ R ₁₄ is obtained from NHR. _O by reaction with isocyanate, for example HN = C = O or R ₁₀ N = C = O; -NR ₁₀ C (O) R ₃ is obtained from -NHR ₁₀ by reaction with C 1 -C (O) R ₃ in pyridine; C (= NR, _O ) ₁₃ R ₁₄ is obtained from -C (NR, ₃ R ₁₄ ) SR ₃ with H ₃ NR ₃ OA _c 'by heating in alcohol; C (NR ₁₃ R ₁₄ ) SR ₃ is obtained from -C (S) NR ₁₃ R ₁₄ with R ₆ -I in an inert solvent, e.g. acetone; C (S) NR ₁₃ R ₁₄ (such as R 1 or R ₄ is not hydrogen) is obtained from C 1 S) NH _; c HNR _I3 R ₁₄ ;

-C (= NCN) -NR ₁₃ R ₁₄ is obtained from -C (= NR _; R ₁₄ ) -SR _{3 with} NH ₂ CN by heating in anhydrous alcohol, alternatively from -C (= NH) \ R ₁₃ R _{| 4} by reaction with BrCN and NaOEt in EtOH: -NR ₁₀ -C (= NCN) SR ₈ is obtained from NHR ₁₀ interaction with (R ₈ S) ₂ C = NCN;

-NR. _o S ₂ R ₃ is obtained from -NHR ₁₀ by reaction with NR ₁₀ C (O) R ₈ by reaction with a Lawesson reagent [2,4-bis- (4-methoxyphenyl) -1,3,2,4-dithiadiphosphetane -2,4-disulfide]:

-NR. ₀ S0 ₂ CF ₃ is obtained from NHR ₆ with triflic anhydride and base wherein R _3, R _6, R _10, R ₁₃ and R ₁₄ are as defined in formula I herein.

The precursors of the groups R 1, R ₂ and R ₄ may be other groups R _p R ₂ and R ₄ , which can be transformed using standard functional group conversion techniques. For example a compound of formula I, wherein R ₂ is substituted with halo, _C10 alkyl, can be transformed into the corresponding C ₁₀ alkyl N ₃ derivative by reacting with a suitable azide salt, and thereafter if desired can be reduced to the corresponding A C ₁₀ alkyl NH ₂ compound that can react with R ₁₈ S (O) ₂ X, with X being halogen (e.g. chlorine) to give the corresponding C ₁₁₀ alkyl NHS (O) ₂ R ₁₈ compound.

An alternative compound of formula I in which R, substituted by halogen C ₁₁₀ alkyl, may be reacted with an amine R ₁₃ R ₁₄ N to give the corresponding C ₁₁₀ alkyl NR ₁₃ R ₁₄ , or may be reacted with an alkali metal salt of R ₁₈ SN. to give the corresponding C ₁₁₀ alkyl SR _s compound.

According to Scheme I, the compounds of formula I are prepared by reacting a compound of formula II with a compound of formula III in which p is 0 or 2, R 35

R ₂ and R ₄ are as defined above in formula I or are the precursors of the groups R _p R ₂ and R ₄ and Ar is optionally substituted phenyl group and then, if necessary, the conversion of the precursors R _p R ₂ and R ₄ in the groups R _p R ₂ and R ₄ .

It is recommended that the reaction be carried out at room temperature or under cooling (eg -50 ° C to 10 ° C) or heated in an inert solvent such as methylene chloride, DMF, tetrahydrofuran, toluene, acetonitrile or dimethoxyethane in the presence of a suitable base such as 1, 8-diazabicyclo [5.4.0] undec7-ene (DBU) or guanidine base such as 1,5,7-triaza-bicyclo [4.4.0] dec-5-ene (TBD).

It has been found that the intermediates of formula II are very stable and can be stored for a long time. Preferably p is 2. PTC is defined as a phase transfer catalyst.

The compounds of formula II have a structure

Ar S (O) _p

wherein p is 0 or 2; R ₄ is as defined in formula I and A ₂ is optionally substituted aryl as defined herein.

It is recommended that Ar be phenyl optionally substituted by C ₁₄ alkyl, C _1-4 alkoxy or halogen. Preferably Ar is phenyl or 4-methylphenyl, for example a tosyl derivative.

The interaction of a compound of formula II in which p is 2 with a compound of formula III in Scheme I yields substantially higher yields of a compound of formula I than that of which p is 0. In addition, the reaction of the compounds of formula II, in which p is 2, is more environmentally and economically acceptable. When p is 0, the preferred solvent used is methylene chloride, which is not environmentally friendly in many processes, and the preferred base, TBD, is also expensive, resulting in age products and impurities compared to the synthesis used (p is 2), as described here.

In Scheme I, 1,3-dipolar cyclodealdehyde anion of substituted thiomethylisocyanide (when p is 0) to imine is used. In particular, this interaction requires a strong base, such as an aminobase, for the deprotonation step. Commercially available TBD is preferred, although tert-butoxide, Li ⁺ or Na ⁺ , or K ⁺ hexamethyldisilazide may also be used. Although methylene chloride is the preferred solvent, other halogenated solvents such as chloroform or carbon tetrachloride may be used; ethers such as THF, DME, DMF, diethyl ether, tert-butylmethyl ether; as well as acetonitrile, toluene or mixtures thereof. The reaction may be carried out at a temperature of from about -20 ° C to about 40 ° C, preferably from 0 ° C to 23 ° C, more preferably from 0 ° C to 10C and most preferably at 4 ° C for reaction reactions insertion of the BC group into the pyrimidine.

In another embodiment, the compounds of formula I may be prepared by coupling a suitable derivative of a compound of formula IX

in which T ₁ is hydrogen and T ₄ is R ₄ or alternatively, T ₁ is R ₄ is T ₄ is hydrogen, where R 1, R ₂ , and R ₄ are defined above by:

(i) when T, is hydrogen, a suitable derivative of the heteroaryl ring R 1, under conditions of ring formation, by coupling the heteroaryl ring R to the imidazole nucleus at the 5-position; and (ii) when T ₄ is hydrogen, a suitable derivative of the aryl ring R ₄ H, under conditions of ring attachment, the bonding of the aryl ring R ₄ to the imidazole nucleus at the 4 position is carried out.

Such aryl / heteroaryl coupling reactions are well known in the art. In general, the organometallic synthetic equivalent of the anion of one component is coupled to a reactive derivative of the other component in the presence of a suitable catalyst. The anionic equivalent may be formed either from the imidazole of formula IX, in which case the aryl / heteroaryl compound provides the reactive derivative, or from the aryl / heteroaryl compound, in which case imidazole provides the reactive derivative. Accordingly, suitable derivatives of a compound are Formula IX or the aryl / heteroaryl ring are organometallic derivatives such as organomagnesium. organotin zinc, organotin, and boric acid derivatives, and suitable reactive derivatives are bromine, iodine, trifluorosulfonate and trifluoromethane sulphonate derivatives. Suitable procedures are described in WO 91/19497 _d

Suitable organomagnesium and organotin zinc derivatives of formula IX may be reacted with a halogen, fluorosulfonate or triflate derivative of the heteroaryl or aryl ring in the presence of a ring coupling catalyst such as palladium 0 or palladium II catalyst following the procedure of Kumada et al. , Tetrahedron Letters, 22, 5319 (1981). Such suitable catalysts are tetrakis- (triphenylphosphine) -palladium and PdCl ₂ [1,4-6HcCn-6Hiphosphino) -butane], optionally in the presence of lithium chloride and a base such as triethylamine. In addition, a nickel II catalyst such as Ni (II) C1 ₂ (1,2-biphenylphosphino) ethane can be used to bond the aryl ring, following the procedure of Pridgen et al., J. Org.Chem., 1982 , 47, 4319. A suitable reaction solvent is hexamethylphosphoramide. Suitable derivatives, when the aryl ring is phenyl, are bromine, trifluorosulfonate, triflate and preferably iodine derivatives. Organomagnesium and organotin zinc derivatives can be prepared by reacting a compound of formula IX or a bromine derivative with an alkyl lithium compound to give the corresponding lithium compound by deprotonation or transmetallization, respectively. This lithium intermediate may then be reacted with excess magnesium halide or zinc halide to give the corresponding organometallic reagent.

A trialkyl tin derivative of a compound of formula IX may be reacted with a bromine, trifluorosulfonate, triflate, or preferably iodine derivative of an aryl or heteroaryl cyclic compound, in an inert solvent such as tetrahydrofuran, preferably containing 10% hexamethylphosphate palladium (O) catalyst, for example tetrakis- (triphenylphosphine) palladium, by the method described by Stille, J.Amer. Chem Soc., 1987, 109, 5478, US 4 719 218 and 5 002 942, or using a palladium II catalyst in the presence of lithium chloride, optionally by adding a base such as triethylamine, in an inert solvent such as dimethylformamide. The trialkyl tin derivative can be obtained in the usual way by metallizing the corresponding compound of formula IX with a lithium agent such as sec-butyllithium or norm-butyllithium in an ether solvent such as tetrahydrofuran, or reacting the bromine derivative of the corresponding compound of formula IX with alkylthio, and then, in each case, reaction with a trialkyl tin halide. Alternatively, the bromine derivative of the compound of formula IX may be reacted with a suitable heteroaryl or aryl trialkyl tin compound in the presence of a catalyst such as tetrakis- (triphenylphosphine) -palladium under conditions similar to those described above.

Bromic acid derivatives may also be used. Hence, a suitable derivative of a compound of formula IX as a bromine, iodine, triflate or fluorosulfonate derivative can be reacted with a heteroaryl or arylboronic acid in the presence of a palladium catalyst such as tetrakis- (triphenylphosphine) palladium or PcCl ₂ [1,4-biC1] - (diphenylphosphino) -butane] in the presence of a base such as sodium bicarbonate, under reflux conditions, in a solvent such as dimethoxyethane (see Fischer and Haviniga, Rec. Trav.Chim Pays Bas, 84, 439, 1965, Snieckus, V., Tetrahwdron. Lett., 29, 2135, 1988 and Terashimia, M., Chem. Pharm. Bull., 11, 4755,

1985). It can be operated in the absence of water, for example using a solvent such as DMF, at a temperature of about 100 ° C, in the presence of a Pdll catalyst (see Thompson WJetal. J. Org. Chem., 49, 5237, 1984). . Suitable boric acid derivatives may be prepared by reacting a magnesium or lithium derivative with a trialkylborate ester such as triethyl-, triisopropyl- or tributylborate, by a standard procedure.

In such coupling reactions, it can easily be appreciated that the functional groups present in the compounds of formula IX should be considered. Thus, in general, amino substituents and sulfur substitutes should not be oxidized or protected.

The compounds of formula IX are imidazoles and can be prepared according to any of the schemes described above for the preparation of compounds of formula I. In particular, α-halo-ketone or other suitably activated ketones R ₄ COCH ₂ Hal ( for compounds of formula IX, wherein T is hydrogen) or R ^ OCHjHal (for compounds of formula IX, wherein T ₄ is hydrogen) may be reacted with an amidine of the formula R ₂ NH-C = NH, wherein R ₂ is defined in formula I, or a salt thereof, in an inert solvent such as a halogenated hydrocarbon solvent, e.g. chloroform, at a moderate increase in temperature. minutes and, if necessary, in the presence of a suitable condensing agent such as a base. The preparation of suitable α-halogen ketones is described in WO91 / 19497A ₁ . Suitable reactive esters are esters of strong organic acids such as lower alkanesulfonic or arylsulfonic acid, e.g. methane or p-toluenesulfonic acid. Amidine is preferably used as a suitable hydrochloride salt, which can then be converted to free amidine in situ, using a two-phase system in which the reactive ester is in an inert organic solvent such as chloroform and the salt is in the aqueous phase to which an aqueous solution of the base, in a dimolar amount, is slowly added with vigorous stirring. Suitable amidines can be prepared by standard methods, see, for example, Garigipati R., Tetrahedron Letters, 190, 31, 1989.

Compounds of formula I may be prepared by a method which consists of reacting a compound of formula IX in which T is hydrogen with an N-acylheteroaryl salt according to the method described in US 4 803 279, US 4 719 218 and US 5 002 942 to give an intermediate wherein the heteroaryl ring is attached to the imidazole nucleus and is present as its 1,4-dihydro derivative, which intermediate may then be subjected to oxidative deacylation conditions ( Scheme II).

The heteroaryl salt, for example the pyridinium salt, may be either pre-prepared or preferably obtained in situ by the addition of a suitable carbonyl halide (such as acyl halide, aroyl halide, arylalkyl halogenformate ester, or, preferably, alkylhalogenoformate benzeneformate benzeneformate benzeneformate benzeneformate benzeneformate benzeneformate benzeneformate benzeneformatebenzoateformyl) or preferably ethyl chloroformate) to a solution of a compound of formula IX of the heteroaryl compound RH, or in an inert solvent such as methylene chloride to which heteroaryl is added o compound. Suitable deacylating and oxidizing conditions are described in US 4 803 279, 4 719 218, and 5002 942.

Suitable oxidation systems include sulfur in an inert solvent or a mixture of solvents such as decalin, decalin and diglyme, p-cement, xylene or mesitylene, under reflux conditions, or preferably potassium tert-butoxide in tert-butanol in dry air or oxygen.

SCHEME II

In another embodiment of the process illustrated in Scheme III below, compounds of formula I may be prepared by reacting a compound of formula X upon heating or by the addition of a cyclizing agent such as phosphorus oxychloride or phosphorus pentachloride (see Engel and Steglich, Liebids Ann, Chem., 1978, 1916 and Strzybny et al., J. Org. Chem., 1963, 28, 3381). The compounds of formula X can be taught, for example, by acylation of the corresponding α-ketoamine with an activated formate derivative such as the corresponding anhydride, under standard acylation conditions and subsequent imine formation with R ₂ NH _r The amino ketone can be obtained from the starting ketone by oxidation and reduction and the necessary ketone can be obtained by decarboxylation of the β-ketoester obtained by the condensation of an aryl (heteroaryl) acetic ester with the R, COX component.

SCHEME III

Scheme IV below lists two 2 different ways in which a ketone (Formula XI) is used to prepare a compound of Formula I. The heterocyclic ketone XI is prepared by the addition of an alkylheterocycle anion such as 4-methylquinoline (obtained by its interaction with alkyl lithium as normal.

butyllithium) to an N-alkyl-O-alkoxybenzamide ester or any other suitably activated derivative in the same oxidized state.

Alternatively, the anion may be fused to benzaldehyde to give an alcohol, which is then oxidized to ketone XI.

SCHEME IV

r ^ nhr ₂ q N — Ro R4 0 X ^ 1 \ j ^ CHj'Li ⁴ · Ί Br ₂ rAo * rAo HN Ri \ ^ r || 1 NH R4 0 Λ x>

In another embodiment of the method, an N-substituted compound of formula I can be obtained by reacting the anion of an amide of formula XII

RjCHjNRjCOH xn in which R 1 and R ₂ are defined above with:

a) nitrile of formula XIII

R.CN XIII wherein R ₄ is as defined above, or

B) an excess of alkyl halide, for example acyl chloride of formula XIV

R.COHal XIV in which R ₄ is defined above and Hal is halogen or the corresponding anhydride to give a bis-acylated intermediate which is then reacted with an ammonia source such as ammonium acetate.

base ^R 4 / ^a

--►

(XI)

One embodiment of this approach is illustrated in Scheme V. A primary amine (R ₂ NH ₂ ) is reacted with a halogenmethyl heterocycle R 1 H 2 X to give a secondary amine which is converted into an amide in a known manner. Alternatively, the amide may be prepared, as described in Scheme V, by alkylation of the formamide R 1 H 2 X. Deprotonation of this amide with a strong amide base such as lithium diisopropylamide or sodium bis- (trimethylsilyl) amide, followed by the addition of excess aroyl chloride, yields a bisacylated compound, which is then cyclized to imidazole of formula I, acetic acid containing ammonium acetate. Alternatively, the amide anion may be reacted with substituted arylnitrile to give imidazole of formula I.

The description and the example schemes illustrate the method (as in Scheme I above). Various pyrimidinaldehyde derivatives 6 and 7, as indicated in Scheme VI below, can be prepared by modifying the technologies of Bredereck et al. (Chem. Ber. 1964, 97, 3407). These pyrimidinaldehydes are then used as intermediates for the syntheses described below.

SCHEME VI

The reaction of imines with tosylmethylisonitrile was first commented on by Leusen (van Leusen et al., J. Org. Chem., 1977, 42, 1153). The following conditions are given: tert-butylamine (TpeTBuNH ₂ ) in dimethoxyethane (DME), K ₂ CO ₃ in MeOH and NaH in DME. Reproducing these examples turns out to yield low yields. The second method, involving an amine exchange reaction to produce tert-butylamine followed by reaction with isocyanide to give 1-tertiary imidazole, is also feasible. It is appropriate to use any primary amine as a base. Secondary amines, although not preferred, can also be used, can decompose isonitriles slowly. The reaction requires about 3 equivalents of amine to complete the reaction, yielding approximately 50% isolated yield. Secondary amines (diisopropylamine), when used, act very slowly and are not as effective. The tertiary and aromatic amines used, such as pyridine and triethylamine, did not interact under some of the conditions tested, but more basic species such as DBU and 4-dimethylaminopyridine (DMAP), although to a small extent, produced the desired product and could thus be suitable for application.

As indicated in Schemes VII and VIII below, the pyrimidinaldehyde of Scheme VI can be condensed with a primary amine to give an imine that can be easily isolated or reacted in situ with the desired isonitrile in the presence of various suitable bases and solvents , as described herein to give 5- (4-pyrimidinyl) substituted imidazoles, wherein R ₂ and R ₄ have the meanings defined for the compounds of formula I.

One preferred method of preparing compounds of formula I is shown in Scheme VII. The imine obtained and isolated in a separate step is often resinous, which makes it difficult to process. Black is often transferred to the final product. The yield for imine preparation varies, and solvents such as CH ₂ Cl ₂ are often used less frequently to produce environmental imines.

An interaction in which p is 2 requires a suitable base to allow the reaction to proceed. Sufficient base forces are required for the reaction to deprotonate isonitrile. Suitable bases are amine, carbonate, hydride or alkyl or aryl lithium reagent; or mixtures thereof. The bases include (but not limited to) potassium carbonate, sodium carbonate, primary and secondary amines such as morpholine, piperidine. pyrrolidine and other non-nucleophilic bases.

Suitable solvents include, but are not limited to, Ν, Ν-dimethylformamide (DMF), NieCN, halogenated solvents such as methylene chloride or chloroform, tetrahydrofuran 17HF), dimethyl sulfoxide (DMSO), alcohols such as methanol or ethanol, benzene, or benzene or η DME. Preferably the solvent is DMF, DME, THF or MeCN, most preferably DMF. Isolation of the product may generally be clogged with the addition of water or filtration of the product as a pure compound.

Although not suitable for large scale operation, the addition of NaH to isonitrile at temperatures below 25 ° C (in THF) would be beneficial. In addition, BuLi is also known as an effective base for deprotonation of tosylbenzylisonitriles at -50 ° C (DiSanto et al., Synth. Commun., 1995, 25, 795).

Different deprotonation temperature conditions may be used depending on the preferred base. For example, tertBuNH ₂ / DME, K ₂ CO ₃ / MeOH, K ₂ CO ₃ in DMF, at temperatures above 40 ° C, the yield may drop to about 20%, but a small difference is expected between 0 ° C and 25 ° C. . Therefore, temperatures below 0 ° C and above 80 ° C are also contemplated within the scope of the present invention. Preferably the temperature ranges are from 0 ° C to

25C.

As shown in Scheme VIII below, the imine is preferably formed in situ in a solvent. This preferred synthesis is carried out in a single reactor. It is recommended that, when using a primary amine as a salt, the reaction includes a base such as potassium carbonate before the addition of isonitrile. Alternatively, it may be required that the piperidine nitrogen atom be protected as shown below. Reaction conditions such as solvent, base, temperatures, etc. are similar to those mentioned above for the isolation of the imine, as shown in Scheme VII. Under certain conditions, the in situ imine formed may require dehydration conditions or may require acid catalysis.

SCHEME VIII

Scheme IX describes an alternative method for the preparation of compounds of formula I. In this particular case, the alkylthio residue is oxidized to an alkylsulfinyl or alkylsulfonyl residue which is reacted with a suitable alkoxy residue.

SCHEME IX

Another embodiment of the invention is a new hydrolysis of 2-thiomethylpyrimidinacetal to 2-thiomethylpyrimidinaldehyde, as shown in Scheme X below. Hydrolysis of acetal to aldehyde using various known reaction conditions, such as formic acid, did not result in satisfactory aldehyde yields <13%. Preferred synthesis involves the use of AcOH (freshly prepared) as solvent and concentrated sulfuric acid under heating conditions, preferably cat40 lytic. The heating conditions include temperatures from about 60 to 85 ° C, preferably from 70 to 80 ° C, with higher temperatures darkening the reaction mixture. After the reaction is complete, the mixture is cooled to approximately room temperature and the acetic acid is removed.

A more preferred embodiment involves heating the acetal in 3N HCl at 40 ° C for 18 h, cooling and extracting the hydrogenated carbonate solution in EtOAc.

SCHEME X

Although in the schemes described herein, for example, optionally with a piperidine radical for R ₂ or 4-fluorophenyl for R ₄ , any suitable R or R ₄ may be thus incorporated if obtained on the basis of a primary amine. Alternatively, any suitable R ₄ may be added by isonitrile.

The compounds of formula II in Scheme I can be prepared by the method of Leusen et al., Supra. For example, compounds of formula II can be prepared by dehydrating a compound of formula IV - Scheme I, with Ar, R ₄ and p having the above meanings. Suitable dehydrating agents are phosphorus oxychloride, oxalyl chloride, thionyl chloride, phosgene or tosyl chloride in the presence of a suitable base such as triethylamine or diisopropylethylamine, or the like, such as pyridine. Suitable solvents are dimethoxyether, tetrahydrofuran or halogenated solvents, preferably THF. The reaction is most effective when the reaction temperature is maintained between -10 ° C and 0 ° C. At lower temperatures the reaction is incomplete, and at higher temperatures the solution darkens and the yield of the product decreases.

The compounds of formula IV - Scheme I can be prepared by reacting a compound of formula V - Scheme I, R ₄ CHO, wherein R ₄ is defined herein with ArS (O) _p H and formamide, with or without removal of water , preferably under dehydration conditions, at room temperature or at 20 elevated temperatures, e.g. 30 ° C to 150 ° C, usually refluxed, optionally in the presence of an acid catalyst. Alternatively, trimethylsilyl chloride may be used in place of the acid catalyst. Examples of acid catalysts are camphor-10-sulfonic acid, formic acid, p-toluenesulfonic acid, hydrochloric acid or sulfuric acid.

The most advantageous process for preparing isonitrile of formula II is illustrated below in Scheme XI.

The conversion of the substituted aldehyde to tosylbenzyl formamide can be accomplished by heating the aldehyde, 1-Scheme XI, with an acid such as p-toluenesulfonic acid, formic acid or camphorsulfonic acid; with formamide and toluenesulfinic acid [under reaction conditions of about 60 ° C for about 24 hours]. Preferably, no solvent is used. The reaction can be carried out in low yield (<30%) when solvents such as DMF, DMSO, toluene, acetonitrile or excess formamide are used. Temperatures below 60 ° C are generally unfavorable to obtain the desired product, and temperatures above 60 ° C may give rise to a product which decomposes or results in benzyl bisformamide, 2-Scheme XI.

Tosylbenzylformamide compound can be prepared by reacting the bisformamide intermediate, 2-Scheme XI with p-toluenesulfinic acid. In this preferred embodiment, the preparation of bisformamide from the aldehyde is accomplished by heating the formamide aldehyde in a suitable solvent with an acid catalyst. Suitable solvents are toluene, acetonitrile, DMF and DMSO or mixtures thereof. Acid catalysts are those well known in the art and include (but are not limited to) hydrogen chloride, p-toluenesulfonic acid, camhosulfonic acid, and other anhydrous acids. The reaction may be carried out at temperatures in the range of about 25 ° C to 110 ° C, preferably about 50 ° C, for about 4 to about 5 hours, with longer reaction times also being acceptable. Decomposition of the product or lower yields may occur at elevated temperatures (<70 ° C) over a long reaction time. Removal of water from the reaction mixture is necessary for complete conversion of the product.

Preferred conditions for the conversion of the bis-formamide derivative to tosylbenzylformamide are carried out by heating the bisformamide in a suitable solvent with an acid catalyst and p-toluenesulfinic acid. The solvents used for this interaction include (but are not limited to) toluene and acetonitrile or mixtures thereof. Other mixtures of these solutions with DMF or DMSO may also be used, but lower yields may be obtained. The temperature ranges from about 30 ° C to about 100 ° C. Temperatures lower than 40 ° C and higher than 60 ° C are not recommended as yield and speed decrease. A preferred range is 40 to 60 ° C, especially about 50 ° C. The optimum time is about 4 to 5 hours, although it may be longer. Preferably the acids used include (but are not limited to) toluenesulfonic acid, camphorsulfonic acid and hydrochloric acid and other anhydrous acids. It is particularly preferred that the bisformamide be heated in a 1: 1 mixture of toluene: acetonitrile with p-toluenesulfinic acid and hydrogen chloride.

The preferred embodiment for the synthesis of a tosylbenzylformamide compound that is carried out in a single reactor. In this method, the aldehyde is initially converted to a bisformamide derivative, after which the bisformamide derivative is reacted with toluenesulfonic acid. This procedure combines optimal conditions in a single, effective process. In this way high yields (<90%) of arylbenzylformamide are realized.

A catalyst such as trimethylsilyl chloride (TMSCl), in a preferred solvent toluene: acetonitrile in a 1: 1 ratio, is used under the preferred reaction conditions. A reagent such as TMSC1 is preferred because it reacts with the resulting water and at the same time releases hydrogen chloride to catalyze the reaction. The use of hydrogen chloride and p-toluenesulfonic acid is also preferred. Therefore, three recommended reaction conditions are included: 1) use of a dehydrating agent that provides hydrogen chloride together with TMSC1; or 2) use of a suitable dehydrating agent and a suitable source of acid such as (but not limited to) camphorsulfonic acid, hydrochloric acid or toluenesulfonic acid; and 3) alternative dehydrating conditions such as azeotropic removal of water and the use of an acid catalyst and phthalenesulfinic acid.

Compounds of formula II in which p is 2 can be prepared by reacting in the presence of a strong base with a compound of formula VI - Scheme I, R ₄ CH ₂ NC with a compound of Formula VII - Scheme I, ArSO ₂ L _p as R ₄ and Ar are as defined and L is a leaving group such as halogen, for example fluorine. Suitable strong bases include (but are not limited to), alkyllithium such as butyllithium or lithium diisopropylamide (Van Leusen et al., Tetrahedron Letters, No. 23, 2367-68 (1972)).

Compounds of formula VI - Scheme I can be prepared by reacting a compound of formula VIII - Scheme I, R ₄ CH ₂ NH ₂ with alkyl formate (eg ethyl formate) to give the intermediate amide, which can be converted to the desired isonitrile by reacting with a known dehydrating agent such as (but not limited to) oxalyl chloride, phosphorus oxychloride or tosyl chloride in the presence of a suitable base such as triethylamine.

Alternatively, a compound of formula VIII of scheme I may be transformed into a compound of formula VI of scheme I by reaction with chloroform and sodium hydroxide in aqueous dichloromethane in the presence of a phase transfer catalyst.

The compounds of formula III - Scheme I can be prepared by reacting a compound of formula R 1 HO with a primary amine R ₂ NH ₂ .

The amino compounds of formula VIII - Scheme I are known or may be prepared from the corresponding alcohols, oximes or amides using standard conversion of functional groups.

Suitable protecting groups for use with hydroxyl groups and imidazole nitrogen are well known to those skilled in the art and have been described in many literature, for example, Protective Groups in Organic Synthesis, Greene TW, Wiley-Interscience, New York. 1981. Suitable examples of protecting hydroxyl groups are silyl ethers such as tert-butyldimethyl or tert-butyldiphenyl and alkyl ethers such as methyl bonded via a variable chain alkyl chain (CR ₁₀ R ₂₀ ) _n . Suitable examples of protecting groups for imidazole nitrogen are tetrahydropyranyl.

Pharmaceutically acceptable acid addition salts of compounds of formula I may be prepared in a known manner, for example, by treatment with an appropriate amount of acid in the presence of a suitable solvent.

Examples of application and implementation of the invention

Treatment methods

The compounds of formula I or their pharmaceutically acceptable salts may be used to produce drugs for the prophylaxis or treatment of any disease condition in a human or other mammal that is exacerbated or caused by excessive or unregulated cytokine production by the cells of that mammal, such as monocytes and / or macrophages, not limited to.

The compounds of formula I suppress inflammatory cytokines, such as IL-1, IL-6, IL-8 and TNF, and are therefore useful as medicaments. IL-1, IL6. IL-8 and TNF affect a large number of cells and tissues and these cytokines, as well as other leukocyte-produced cytokines, are important and critical mediators of inflammation in a wide variety of disease states and conditions. Inhibition of these inflammatory cytokines is useful for controlling, limiting and ameliorating these disease states.

The invention also relates to a method of treating diseases with a cytokine mediator, which comprises administering an effective amount of a cytokine to a compound of formula I or a pharmaceutically acceptable salt thereof.

The compounds of formula I are able to inhibit excitable pro-inflammatory proteins such as COX-2, also known by various names, such as prostaglandin endoperoxide synthase-2 (PGHS-2), and are therefore useful as medicaments. These pro-inflammatory lipid mediators of the cyclooxygenase pathway (CO) are produced by the induced COX-2 enzyme. Therefore, the regulation of COX-2, which causes the production of arachidonic acid products such as prostaglandins, affects a large number of cells and tissues and is an important mediator of inflammation in various disease states. Expression of COX-2 is not accomplished by the compounds of formula I. This selective inhibition of COX-2 attenuates or attenuates the ulcerative action associated with the inhibition of COX-1, thereby inhibiting prostaglandins, which are important cytoprotectants. Thus, inhibition of these pro-inflammatory mediators is applied to control, reduce and alleviate many of these disease states. Of particular note is that these inflammatory mediators, in particular prostaglandins, influence pain or edema by sensitizing pain receptors. This aspect of influencing pain, therefore, involves the treatment of neuromuscular pain, headache, tumor-related pain, and arthritis pain. The compounds of formula I or their pharmaceutically acceptable salts are used for the prophylaxis or treatment of humans or other mammals by inhibiting the synthesis of the COX-2 enzyme.

The invention also relates to a method of inhibiting the synthesis of COX-2, which comprises administering an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

The invention also relates to a method for the prophylactic treatment of humans or other mammals by inhibiting the synthesis of COX-2 enzyme.

In particular, the compounds of formula I or their pharmaceutically acceptable salts are used for the prophylaxis or treatment of any disease condition in mammals that is exacerbated or caused by excessive or unregulated production of IL-1, IL-8 or TNF by cells, including , but not limited to, monocytes and / or macrophages.

In another aspect, the invention relates to a method of inhibiting the production of IL-1 in a mammal, which comprises treatment with an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

There are a number of diseases in which excessive or unregulated production of IL-1 affects the exacerbation or causing of the disease. These include rheumatoid arthritis, osteoarthritis, stroke, endotoxemia or toxic shock syndrome, other acute or chronic inflammatory conditions such as endotoxin-induced inflammatory response or inflammatory bowel disease, tuberculosis, atherosclerosclerosis, mucosa, bones, psoriasis arthritis, Reiter's sin20 dromo, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis and acute synovitis. New studies have also linked IL-1 action to diabetes, pancreatic β-cells and Alzheimer's disease.

The invention also relates to a method of inhibiting the production of TNF in mammals, which method comprises treating with an effective amount of a compound of formula I 10 or a pharmaceutically acceptable salt thereof.

Excessive or unregulated TNF production is involved in the mediation or exacerbation of many diseases, including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions, sepsis, septic shock, endotoxic shock syndrome, gramotomy syndrome respiratory distress in adults, stroke, cerebral malaria, chronic pulmonary inflammatory disease. silicosis, pulmonary sarcoisosis, bone resorption disease such as osteoporosis, reperfusion injury, transplant / receiver response, allograft reaction, fever and myalgia due to infection such as influenza, cachexia due to infection or malnutrition, AIDS), AIDS (AIDS), spin-related complex (ARC), keloid formation, cicatrix tissue formation, Crohn's disease, ulcerative colitis and pyresis.

The compounds of formula I are also used to treat viral infections in viruses that are susceptible to regulation by TNF or abolish TNF production in vivo. Such viruses are those that produce TNF as a result of infection, or those that are susceptible to inhibition, thereby diminishing replication, directly or indirectly, by a compound of formula I, inhibiting TNF. Such viruses include (but are not limited to) HIV-1, HIV-2 and HIV-3, cytomegaliovirus (CMV), influenza, adenovirus and Herpes-group viruses such as (but not limited to), Herpes Zoster and Herpes Simplex.

The invention also relates to a method of treating mammals affected by human immunodeficiency virus (HIV), which comprises treating with an effective TNF inhibitory amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

The compounds of formula I may also be used for the veterinary treatment of mammals (other than humans) needing to inhibit TNF production. Diseases with the TNF mediator for treatment, therapeutic or prophylactic, in animals include disease states as noted above, but in particular viral infections. Examples of such viruses include (but are not limited to) lentiviral infections such as equine anemia virus, goat arthritis virus, visna virus, maedi virus, or retroviral infections such as (but not limited to) the immune virus cat failure (FIV), bovine immunodeficiency virus, or canine immunodeficiency virus or other retroviral infections.

The compounds of formula I may also be used topically for the treatment or prophylaxis of disease states caused or exacerbated by excessive cytokine production such as IL-1 or TNF, respectively, inflamed joints, eczema, psoriasis, and other inflammatory conditions of the skin, like sunburn; inflammatory conditions of the eyes, including conjunctivitis; pyresis, pain, and other conditions associated with inflammation.

The compounds of formula I have also been shown to inhibit the production of IL-8 (interleukin-8, NAP).

The invention relates to a method of inhibiting the production of IL-8 in mammals, which consists in treatment with an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof.

Disease conditions exist in which the excessive or unregulated production of IL-8 is involved in exacerbating or causing the disease. These diseases are characterized by massive neutrophil infiltration such as psoriasis, bowel inflammation, asthma, cardiac or renal reperfusion injury, adult respiratory distress syndrome, thrombosis and glomerulonephritis. All of these conditions are associated with increased production of IL-8, which causes the chemotaxis of neutrophils to the site of inflammation. In contrast to other inflammatory cytokines (IL-1, TNF, and IL-6), IL-8 has the unique property of accelerating the chemotaxis and activation of neutrophils. Due to this, inhibition of IL-8 production results in a direct decrease in neutrophil infiltration.

The compounds of formula I are administered in an amount sufficient to inhibit the production of the cytokine, in particular IL-1, IL-6, IL-8 or TNF, so that it is regulated to a normal level and to improve or prevent the disease state . Abnormal levels of IL-1, IL-6, IL-8 or TNF, for example in the context of the present invention, are:

(i) concentrations of free (non-cell bound) IL-1, IL-6, IL-8 or TNF greater than or equal to 1 pg / ml;

(ii) any cell-bound IL-1, IL-6, IL-8 or TNF; or (iii) the presence of IL-1, IL-6, IL-8 or TNF mRNA above baseline concentrations in cells or tissues in which IL1 is produced. IL-6, IL-8 or TNF, respectively.

The finding that the compounds of formula I are inhibitors of cytokines, in particular IL-1, IL-6, IL-8 or TNF, is based on the effects of the compounds of formula I on the production of IL-1, IL- 8 or TNF in the in vitro assays described below.

As used herein, the term "inhibition of IL-1 production (IL-6, IL-8 or TNF)" refers to:

a) reducing the extremely high in vivo cytokine concentration (IL-1, IL-6, IL-8 or TNF) in humans to normal or below normal levels by inhibiting in vivo cytokine release in all cells, including (without limitation) only), monocytes and macrophages;

b) regulating the genomic level of excessive in vivo cytokine concentrations (IL-1, IL-6, IL-8 or TNF) in humans to normal or below normal levels;

c) regulation by inhibition of direct cytokine synthesis (IL-1, IL-6, IL-8 or TNF) as a posttranslational action; or

d) regulating the translational level of extremely high in vivo concentrations of cytokines (IL-1, IL-6, IL-8 or TNF) in humans to normal or below normal concentrations.

As used herein, the term "disease or disease state with a TNF mediator" refers to any or all disease states in which TNF plays a role, either by producing TNF itself or by TNF causing the release of another monokine such as (but not limited to) IL-1, IL-6, or IL-8. A disease state in which, for example, IL-1 is a major component and whose production or action is enhanced in response to TNF, is therefore considered a disease state with a TNF mediator.

As used herein, the term "cytokine" refers to any produced polypeptide that affects cell function and is a molecule that modulates the interaction between cells in an immune, inflammatory or haematopoietic response. Cytokines include (but are not limited to) monokines and lymphokines, no matter which cells produce them. For example, the monokine is generally considered to be produced and secreted by a mononuclear cell, such as a macrophage and / or monocyte. However, many other cells also produce monokines, such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epidermal keratinocytes, and B lymphocytes. Lymphokines are generally considered to be produced by lymphocyte cells. Examples of cytokines include, but are not limited to, interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor (TNF-α) and tumor necrosis factor -beta (TNF-β).

As used herein, the term "cytokine inhibitory" or "cytokine inhibitory amount" refers to an effective amount of a compound of formula I that causes the in vivo concentrations of the cytokine to fall to normal or below normal when given to a patient for prophylaxis or treating a disease condition that is exacerbated or caused by excessive or unregulated cytokine production.

As used herein, the cytokine referred to in the expression &quot; cytokine inhibition for use in the treatment of an HIV infected person &quot; is a cytokine that is involved in a) initiating and / or maintaining T-cell activation and / or HIV gene. mediator T cell expression and / or replication and / or b) any cytokine mediated disease problem, such as cachexia or muscle degeneration.

Because TNF-β (also known as lymphotoxin) has a close structural homology to TNF-α (also known as cachectin) and because each of them elicits similar biological responses and binds to the same cellular receptor, both TNF-α and TNF-β is inhibited by the compounds of the invention and is therefore generally referred to in the description as "TNF" unless otherwise stated.

Recently, independently in several laboratories. a new member of the MAP kinase family, alternatively named CSBP, p38 or RK, has been identified [see Lee et al., Nature, vol. 300 η (72), 739 - 746 (1994)]. Activation of this novel protein kinase by dual phosphorylation has been observed in various cellular systems after stimulation with a wide range of stimulants such as physicochemical stress and treatment with lipopolysaccharide or proinflammatory cytokines such as interleukin-1 and tumor necrosis factor. Inhibitors of the cytokine biosynthesis of the invention, the compounds of formula I, have been identified as potent and selective inhibitors of CSBP / p38 / RK kinase activity. These inhibitors are used to determine the activation of signaling pathways in the inflammatory response. In particular, for the first time a specific transduction signaling pathway may be attributed to the action of lipopolysaccharide in the production of cytokines in macrophages. In addition to the aforementioned diseases, the treatment of stroke, neurotrauma, cardiac and renal damage, thrombosis, glomerulonephritis, diabetes and pancreatic β cells, multiple sclerosis, muscle degeneration, eczema, psoriasis, and sunburn are also included.

Cytokine inhibitors have been consistently tested in a number of animal models for anti-inflammatory activity. Model systems have been selected to be relatively insensitive to cyclooxygenase inhibitors to reveal the unique activities of cytokine inhibiting agents. Inhibitors have shown significant activity in many such in vivo studies. Most notable is their efficacy in a model of collagenous arthritis and inhibition of TNF production in a model of endotoxic shock. In the last study, a decrease in plasma TNF concentration correlated with survival and protection against endotoxic shock-related mortality. Also of great importance is the efficacy of the compounds in inhibiting bone resorption in a long-embryonic rat embryonic organ system. (Griswold et al., (1988) Arthritis Rheum. 31: 1406-1412; Badger et al., (1989) Circ. Shock 27, 51-61; Votta et al., (1994) in vitro Bone 15, 533- 538; Lee et al., (1993) B Ann. Y. Acad. Sci. 696, 149-170.

To use a compound of formula I or a pharmaceutically acceptable salt thereof as a therapeutic agent, it is usually processed into a pharmaceutical composition according to standard pharmaceutical practice.

The invention also relates to pharmaceutical compositions containing an effective, non-toxic amount of a compound of formula I and a pharmaceutically acceptable carrier or diluent.

The compounds of formula I, their pharmaceutically acceptable salts and the pharmaceutical compositions containing them, can be conveniently administered by any of the commonly used routes of administration, for example orally, topically, parenterally or by inhalation. The compounds of formula I can be administered in conventional dosage forms prepared by mixing the compound of formula I with standard pharmaceutical carriers according to conventional procedures. The compounds of formula I may also be administered in conventional doses in combination with a known second therapeutically active compound. These procedures may include mixing, granulating and tabletting or dissolving the components as is customary for the desired preparation. The form and nature of the pharmaceutically acceptable carrier or diluent is determined by the amount of active ingredient with which it is to be combined, the route of administration and other well known variables. The carrier (s) should be "acceptable" in the sense that they are compatible with the other components of the dosage form and are not harmful to the patient.

The pharmaceutical carrier used may for example be either solid or liquid. Examples of solid carriers are lactose, white clay, sucrose. talc, gelatin, agar, pectin, arabic gum, magnesium stearate, stearic acid and the like. Similarly, the carrier or diluent may include a release retardant component well known to those skilled in the art, such as glyceryl monostearate or glyceryl distearate alone or in combination with wax.

A large number of pharmaceutical forms can be used. Thus, if a solid carrier is used, the preparation can be tableted, placed in hard gelatin capsules, in powder or pallet form, or in the form of pastilles or sugar dragees. The amount of solid carrier varies widely, but is preferably from about 25 mg to 1 g. When using a liquid carrier, the preparation is in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solutions, or non-aqueous liquid suspensions.

The compounds of formula I may be administered topically, i.e. through on-site application. This includes administering the compound of Formula I externally to the epidermis or oral cavity and the addition of the compound into the eye, ear and nose so that it does not substantially enter the bloodstream. In contrast, non-local administration includes oral, intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include a liquid or semi-liquid preparation suitable for penetrating the skin to the site of inflammation, such as liniments, lotions, creams, ointments or pastes and drops suitable for use in the eyes, ears or nose. The active ingredient for topical administration is from 0.001% to 10% by weight, for example from 1% to 2% by weight of the formulation. However, it may also contain 10% by weight, but it is recommended to contain less than 5% by weight, most preferably 0.1 to 1% by weight of the form.

Lotions according to the invention include those suitable for application to the skin or eyes. The eye lotion may contain a sterile aqueous solution, optionally containing a bactericidal agent, and may be prepared by methods similar to those for the preparation of drops. Lotions or liniments for application to the skin may include a drying agent and a skin cooling agent such as alcohol or acetone and / or moisturizers such as glycerol or oil such as castor or peanut oil.

The creams, ointments or pastes according to the invention are semi-solid dosage forms of the active ingredients for external use. They may be prepared by mixing the active ingredients in a finely divided or powdered form, alone or in solution or suspension on an aqueous or non-aqueous basis, using a suitable apparatus, with an oil or non-oil base. The base may contain hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, metal soap; mucous solution; oil of natural origin, such as almond, corn, peanut, castor or olive oil; lanolin or its derivatives or fatty acids such as stearic or oleic acid in combination with an alcohol such as propylene glycol or macrogel. The form may include any suitable surfactant such as an anionic, cationic or nonionic surfactant, such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural rubber, cellulose derivatives or inorganic materials such as silicates and other ingredients such as lanolin may also be used.

The drops according to the invention may comprise sterile aqueous or oily solutions or suspensions of a bactericidal and / or fungicidal agent and / or any other suitable preservative and preferably include a surfactant. The resulting solution can then be clarified by filtration, transferred to a suitable container, which is then sealed and sterilized by autoclaving or sterilizing at 98-10042 for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to the container by aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the droplets are phenylmercurinitrate or acetate (0.002%), benzalkonium chloride (0.01%), and chlorhexidine acetate (0.01%). Suitable solvents for the preparation of an oily solution include glycerol, dilute alcohol and propylene glycol.

The compounds of formula I may be administered parenterally, i. intravenous, intramuscular, subcutaneous, nasal, rectal, vaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Suitable dosage forms for such administration may be prepared by conventional techniques. The compounds of formula I may also be administered by inhalation, nasally or orally. Suitable dosage forms for such administration as aerosol forms or metered dose inhalers may be prepared by conventional techniques.

For all methods of use according to the invention for the compounds of formula I, the daily dosage regimen is preferably from about 0.1 to about 80 mg / kg body weight, preferably from about 0.2 to 30 mg / kg, most preferably from about 0.5 mg to 15 mg. The daily parenteral dose regimen is about 0.1 to about 80 mg / kg of total body weight, preferably about 0.2 to about 30 mg / kg, and most preferably about 0.5 mg to 15 mg. The daily topical dosage regimen is preferably from 0.1 mg to 150 mg administered once to four times, preferably two or three times daily. The daily inhalation dose regimen is preferably from 0.01 mg / kg to about 1 mg / kg per day. It is known to those skilled in the art that the optimal amount and distribution of individual daily doses of the compounds of formula I and their pharmaceutically acceptable salts is determined by the nature and condition of the patient, the form, method and site of administration for each individual patient, and the optimum is determined by conventional techniques. Specialists also evaluate the optimal course of treatment, i. the number of doses of the compound of formula I or the pharmaceutically acceptable salt thereof per day for a specified number of days, which can be confirmed by one skilled in the art using conventional tests to determine the course of treatment.

The novel compounds of formula I may also be used for the veterinary treatment of mammals in need of inhibition of cytokine production. In particular, cytokine mediator diseases for the therapeutic or prophylactic treatment of animals include diseases such as those listed in the "Methods of treatment, in particular for viral infections" section. Examples of such viruses include (but are not limited to) lentiviral infections such as equine infectious anemia virus, goat arthritis virus, visna virus or maedi virus, or retroviral infections such as (but not limited to) , cats immunodeficiency virus (FIV), bovine immunodeficiency virus or dog immunodeficiency virus, or other retroviral infections.

The invention is described with reference to the following biological examples, which are illustrative and do not limit the scope of the present invention.

Biological examples

The cytokine inhibiting action of the compounds of the invention is determined by the following in vitro studies:

Interleukin-1 (IL-1)

Human peripheral blood monocytes are isolated and purified either from fresh blood from donor volunteers or from blood banks according to the procedure of Colotta et al., J. Immunol. 132, 936 (1984). These monocytes (1 x 10 ⁶ ) were placed in 24-well plates at a concentration of 1-2 million / ml per well. The cells were allowed to adhere to the bottom for 2 h, after which the non-adherent cells were removed by gentle washing. The test compounds were then added to the cells for 1 h before the addition of lipopolysaccharide (50 ng / ml) and the cultures were incubated at 37 ° C for a further 24 h. At the end of this period, the clear culture fluid was removed and purified from cells and all fragments. The clear liquid from the culture was then tested immediately for IL-1 biological activity or by the method of Simon et al., J. Immunol. Methods, 84, 85 (1985) (based on the ability of IL-1 to stimulate interleukin-2 producing cell lines (EL-4) to secrete IL-2 together with the A23187 ionophore) or by the method of Lee et al, J. .Immuno Therapy, 6 (I), 1-12 (1990) (ELISA analysis).

The test compound of formula I, Example 1, showed a positive inhibition in the assay.

Tumor necrosis factor (TNF):

Human peripheral blood monocytes are isolated from leukocyte membranes from blood banks, or thrombophoresis residues according to the procedure of Colotta R. et al., J. Immunol. 132 (2), 936 (1984). Monocytes were placed at a density of 1 x 10 ⁶ cells / ml medium / well in 24-well multi-views. The cells were allowed to adhere to the bottom for 1 h, after which the clear liquid was aspirated and fresh medium (1 ml, RPMI-1640, Whitaker Biomedical Products, Whitaker, CA) containing 1% calf germ plus penicillin was added in streptomycin (10 units / ml). The cells were incubated for 45 min in the presence or absence of the test compound at the order of 1 nM-IO mM (the compounds were dissolved in dimethyl sulfoxide / ethanol so that the final solvent concentration in the culture medium was 0.5% dimethyl sulfoxide / 0 , 5% ethanol). Bacterial lipopolysaccharide (E.coli 055: B5 [LPS] by Sigma Chemicals Co) (100 ng / ml in 10 m phosphate buffered saline) was then added and the cultures incubated for 16-18 h at 37 ° C in 5% CO _g incubator. At the end of the incubation period, the clear culture fluid was removed from the cells, centrifuged at 3000 rpm to remove the cell particles. The clear liquid was then analyzed for TNF activity using either radioimmunoassay or ELISA as described in WO 92/10190 and by Becker et al., J. Immunol., 1991, 147, 4307.

IL-1 and TNF inhibitory activity do not appear to correlate with the properties of the compounds of formula I as mediators of inhibition of arachidonic acid metabolism. In addition, the ability to inhibit the production of prostaglandin and / or the synthesis of leukotriene by non-steroidal anti-inflammatory drugs with potent cyclooxygenase and / or lipoxygenase inhibitory activity does not mean that the compound also necessarily inhibits the production of TNF or IL-1 in non-toxic ones.

In vivo TNF analysis:

The above tests are conducted in vitro, but the compounds of formula I can also be tested in an in vivo system as described in:

1) Griswold et al., Drugs Under Exp. and

Clin. Really. XIX (6), 243-248 (1993) or

2) Boehm et al., Journal of Medicinal Chemistry 39, 3929-3937 (1996).

Interleukin-8 (IL-8):

Human umbilical cord endothelial cells (HUVEC) (Cell Systems, Kirland, Wa) were maintained in culture medium supplemented with 15% calf fetal serum and 1% CSHBGF consisting of aFGF and heparin. The cells were then diluted 20-fold before being placed (250 μΐ each) in gelatin-coated 96-well plates. The culture medium is replaced with fresh medium (200 μΐ) before use. A buffer or test compound (25 μединение, at concentrations between 1 and 10 M) was then added to each well in four replicates and the plates incubated for 6 h in a humidified incubator at 37 ° C, in an atmosphere of 5% CO ₂ . At the end of the incubation period, clear fluid was separated and examined for IL-8 concentrations using an IL-8 ELISA instrument obtained from R&D Systems (Mineapolis, MN). All data are presented as mean values (ng / ml) of multiple samples, based on a standard curve. Where appropriate, IC ₅₀ was determined by nonlinear regression analysis.

Cytokine specific protein binding assay.

Radiocompetitive linkage analysis has been developed to provide highly reproducible primary screening for activity structure studies. This assay provides many advantages over conventional bioassays that use freshly isolated human monocytes as a source of cytokine and an ELISA assay to quantify it. In addition to being much easier to analyze, linkage analysis is widely recognized as highly correlated with bioassay results. A specific and reproducible cytokine inhibitor binding assay was developed using a soluble cystosolic fraction of THP-1 cells and a radiolabeled compound. USSN 08/123175 Lee et al., 1993, USSN; Lee et al., PCT 94/10529, 1994 and Lee et al., Nature 300, n (72), 739-746 (Dec. 1994) disclose said method of drug screening to identify compounds that interact and bind with a cytokine specific binding protein (hereinafter CSBP). However, for the purposes herein, the binding protein may be isolated in solution or in immobilized form or may be expressed by genetic engineering on the surface of recombinant host cells as a phage display system or as fusion proteins. Alternatively, whole cells or cytosolic fractions containing CSBP may be used in screening. Regardless of the shape of the binding protein, most of the compounds are contacted with the binding protein under conditions sufficient to form a compound / binding protein complex, and compounds capable of forming, enhancing or inhibiting the complex are found.

A representative final compound of formula I, Examples 1 to 4, exhibits a positive inhibitory activity with IC ₅₀ <50 μΜ in this binding assay.

CSBP kinase assay:

This assay measures CSBP catalyzed transfer of ³² P from [α- ³² P] ATP to the threonine residue in the epidermal growth factor receptor (EGFR) -produced peptide (T669) in the following sequence: KRELVEPLTPS GEAPNQALLR (residues 661-681). (See Gallagher et al., “Regulation of Stress Induced Cytokine Production by Pyridinyl Imidasoles: Inhibition of CSBP Kinase,” BioOrganic & Medicinal Chemistry, to be published 1996).

The kinase reaction mixtures (total volume 30 µl) contain: 25 mM Hepes buffer, pH 7, 5; 10 mM MgCl ₂ ; 170 [mu] M ATP ^<n ; 10 μM Na-ορτοvanadate; 0.4 mM T 669 peptide; and 20-80 ng expressed from yeast, purified CSBP 2 (see Lee et al., Nature 300, π (72) 739-746 (Dec. 1994). Compounds (5 µl of [6X] stem ^<2) ) were incubated pre-enzyme and peptide 20 min on ice to start reactions with ³² P (MgATP). The reaction mixtures were incubated at 30 ° C for 10 min and quenched by the addition of 10 μl of 0.3 M phosphoric acid. The ³² P-labeled peptide was separated on phosphorous cellulose (Watman, p81) filters by dropwise addition of 30 µl of the reaction mixture. The filters were washed 3 times with 75 mM phosphoric acid, followed by 2 washes with H ₂ O and read at ^32P .

The CSBP Km for ATP is determined to be 170 µM. Therefore, the compounds were screened at Km value of ATP.

⁽²⁾ Compounds are usually dissolved in DMSO and diluted in 25 mM Hepes buffer to give a final DMSO concentration of 0.17%.

Representative final compounds of Formula I, Examples 1, 6 and 7 show a positive inhibitory activity of an IC _J0 <50 uM in this binding assay. Example 8 shows IC _JU > 50 µM in this assay.

Analysis of prostaglandin endoperoxide synthase-2 (PGHS-2):

The following study relates to a method for determining the inhibitory effect of compounds of formula I on human PGHS-2 protein expressed in human monocytes stimulated with LPS.

Method: Human peripheral blood monocytes are isolated from leukocyte membranes by centrifugation through Ficoll and Percoll gradients. Cells were seeded at 2 X 10 ⁶ / wells in 24-well plates and allowed to adhere to the base for 1 h in RPMI supplemented with 1% human AB-serum, 20 mM Lglutamine, penicillin-streptomycin and 10 mM Hepes. The compounds were added at various concentrations and incubated at 37 ° C for 10 min. 50 Lg / well LPS (enzyme expression excitation) was added and incubated overnight at 37 ° C. The clear liquid was separated and the cells were washed once with cold PBS. Cells were lysed in 100 μΐ cold lysis buffer (50 mM Tris / HCl pH 7.5, 150 mM NaCl, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, 300 μg / ml DNAse, 0.1 % TRITON X-100, ImM PMSF, 1 mMleupeptin, 1 mM pepstatin). The lysate was centrifuged (10,000 x g for 10 min at 40 ° C) to remove the fragments and the soluble fraction was subjected to SDS PAGE analysis (12% gel). The gel-separated protein was transferred to the nitrocellulose membrane by electrophoretic means for 2 h at 60 V. The membrane was pre-treated in PBS / Tween buffer, the membrane incubated with 1: 2000 dilution from monospecific antiserum to PGHS-2 or 1: 1000 dilution of antiserum to PGHs-Ι in PBS / Tween with 1% BSA for 1 h with continuous shaking. The membrane was washed 3X in PBS / Tween and then incubated with 1: 3000 dilution of horseradish peroxidase conjugated with rabbit antiserum to rabbit Ig (Amersham) in PBS / Tween with 1% BSA for 1 h with continuous shaking . The membrane was then washed 3X in PBS / Tween and an ECL system (Amersham) was used to demonstrate the expression level of prostaglandin endoperoxide synthase-2.

Results: The test compounds were found to be active in this assay (i.e. inhibit LPS-induced PGHS-2 protein expression in the order of magnitude similar to that of inhibiting cytokine production, as noted in said assays ), including 6- (4-fluorophenyl) -2,3-dihydro-5- (4-pyridinyl) imidazole [2,1b] thiazole and dexamethasone.

Several compounds have been tested and found to be inactive (up to uM), including 2- (4-methylsulfinylphenyl) -3- (4-pyridyl) -6,7-dihydro- (5H) -pyrrolo [1,2- a] imidazole; rolipram; phenidone and NDGA.

None of the tested compounds inhibited PGHS-1 or cPLA ₂ protein concentrations in similar experiments.

TNF-α in traumatic brain injury trials.

The present study relates to the study of tumor necrosis factor mRNA expression in specific brain regions that follow experimentally induced lateral fluid percussion traumatic injury (TBI) in rats. Adult Sprague-Dawlev rats (n = 42) were anesthetized with sodium pentobarbital (60 mg / kg ip) and subjected to moderate-lateral fluid percussion brain injury (2.4 atm) centered on the left temporaparietal cortex ) or sham treatment (anesthesia and surgery without damage, n = 18). Animals were killed by decapitation at 1, 6, and 24 h after injury, the brains were removed and samples were prepared from the left (damaged) parietal car (LC), the corresponding areas from the contralateral right cortex (RC), left hippocampus (LH), and right hippocampus (RH). Total RNA was isolated and Northern blot hybridization was performed and quantified for TNF-α positive control RNA (macrophage = 100%). There was a significant increase in TNF-α mRNA expression in LH (104 ± 17% of positive control, p <0.05 compared to sham), LC (105 ± 21%, p <0.05) and LA (69 ± 8%, p <0.01) in the traumatized hemisphere 1 h after injury. Overexpression of HaTNF-α mRNA was also observed in LH (46 ± 8%, p <0.05), LC (30 ± 3%, p <0.01) and LA (32 ± 3%, <0.01 ) at 6 h, which disappears at 24 h after injury. In the contralateral hemisphere, expression of TNF-α mRNA is increased in RNA is increased in RN (46 ± 2%, p <0.01), RC (4 ± 3%) and RA (22 ± 8%) at 1 h and in RH (28 ± 11%), RC (7 ± 5%) and RA (26 ± 6% p <0.05) at 6 h but not at 24 h after injury. No significant changes in TNF-α mRNA expression were observed in any of the 6 brain regions in any of the hemispheres at any time in sham or non-treated animals. The results show that after parasagittal fluid-percussion brain damage, the temporal expression of TNF-α mRNA changes in specific brain regions, including those in the non-traumatized hemisphere. Because TNF-α is able to induce nerve growth factor (NGF) and stimulate the release of other cytokines by activated astrocytes, this post-traumatic change in TNF-α gene expression plays a significant role in both acute and regenerative responses. for CNS injury.

CNS damage model for IL-β mRNA.

This experiment characterized the regional expression of interleukin-ΐβ (IL-β) mRNA in specific brain regions after experimental lateral fluid percussion traumatic brain injury (TBI) in rats. Adult Sprague-Dawley rats (n = 42) were anesthetized with sodium pantobarbital (60 mg / kg, ip) and subjected to moderate-lateral fluid percutaneous brain injury (2.4 atm) centered on the left temporal lobe (p. 18) , or "sham" treatment (anesthesia and surgery without damage). Animals are killed 1, 6, and 24 h after injury, the brain is separated and samples are prepared from tissues of the left (damaged) parietal cortex (LC), corresponding areas in the contralateral right cortex (RC), the cortex adjacent to the damaged parietal cortex (LA), the adjacent area in the right cortex (RA), left hippocampus (LH), and right hippocampus (RH). Total RNA was isolated and Northern blot hybridization was performed and the amount of IL-Ιβ in brain tissue was represented as a percentage of the radioactivity of IL-Ιβ macrophage RNA applied to the same gel. At 1 h after brain damage, there was a significant and noticeable increase in the expression of IL-Ιβ mRNA in LC (20.0 ± 0.7%) of the positive control, n = 6, p <0.05 compared with sham animal) , LH (24.5 ± 0.9%, <0.05) and LA (21.5 ± 3.1%, p <0.05) in the damaged hemisphere, which remains elevated up to 6 h after LC damage ( 4.0 ± 0.4%, n = 6, p <0.05) and LH (5.0 ± 1.3%, p <0.05), No IL expression was observed in sham or untreated animals -Ιβ mRNA in any of the relevant brain regions. These results indicate that after TBI, the temporal expression of IL-Ιβ mRNA is regionally stimulated in specific brain regions. These regional changes in cytokines such as IL1 β play a role in the post-traumatic or regenerative residual effects of brain damage.

Examples of synthesizing compounds

The invention is illustrated by the following examples which serve to illustrate and should not be construed as limiting the scope of the invention. All temperatures are given in Celsius degrees, all solvents are of pure purity and all interactions are carried out in the absence of water and in argon atmosphere, unless otherwise stated. Mass spectral analyzes were performed on a VG Zab mass spectrometer using rapid atomic bombardment, unless otherwise stated. Ή-NMR (hereinafter NMR) spectra were recorded at 250 MHz using a Bruker AM 250 or At 400 spectrometer. The indicated multiplets are: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, and br means broad signal. The designation eq represents the molecular equivalents of the reagent with respect to the parent reagent.

Flash chromatography was performed on Merck Silica gel 60 (230-400 mesh).

Example 1.

5- (2-methoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

a) 2-H-methylthiopyrimidine-4-carboxyaldehyde dimethylacetal

Pyrogrid aldehyde dimethylacetal (60 ml, 459 mmol) and N, N-dimethylformamide dimethylacetal (60 ml, 459 mmol) were stirred together at 100C for 18 h, after which the mixture was cooled.

Methanol (300 ml), thiourea (69.6 g) and sodium methoxide (231 ml, 25% by weight in MeOH) were added to the above mixture and stirred at 70 ° C for 2 h. After cooling, iodomethane (144 ml) was added dropwise and the mixture was stirred for 3 hours at room temperature. After dilution with EtOAc and H ₂ O, the organic phase was separated, dried (Na ₂ SO ₄ ) and concentrated to give the title compound as a brown oil (75.5 g, 82% yield). Ή NHR (CDCl ₃ ): d 8.17, (d, 1H), 6.77 (d, 1H), 5.15, (s, 1H), 3.40, (s, 6H).

b) 2-methoxypyrimidine-4-carboxaldehyde dimethylacetal

The product of Example 1 (a) (5.0 g, 25 mmol) was dissolved in methanol (100 ml), cooled to 4 ° C and a solution of oxonium salt (9.21 g) in water (100) was added dropwise. ml) (T <15 ° C). It was heated to 23 ° C, stirred for 2 h, poured into 10% aqueous NaOH solution (250 ml) and extracted with EtOAc. The extracts were washed with 10% aqueous NaOH solution, dried (Na ₂ SO ₄ ), filtered, concentrated and flash chromatographed (70% hexane / EtOAc) to give 1.66 g (36%) of the title compound . ESP + (Mass Spec) m / s 185 (MH ⁺ ).

c) 2-methoxypyrimidine-4-carboxaldehyde The product of the previous example (0.54 g, 2.93 mmol) was dissolved in 3M HCl (2.17 ml, 6.5 mmol) and stirred at 23 ° C for 23 h. 3 days, cooled to 4 ° C, separated with EtOAc and basified slightly by the addition of (Na ₂ SO ₃ ). Extraction with EtOAc (5 x 40 ml) gave 0.309 g (76%) of the title compound as a white solid. Ή NMR (CDCl ₃ ): d 9.96 (s, 1), 8.78 (d, 1), 7.46 (d, 1) 4.10 (s, 3).

d) 1-tert-butoxycarbonyl-4-aminopiperidine

-tert-butoxycarbonyl piperidin-4-one (commercially available by Lancaster Chem) (39.9 g, 0.20 mol), THF (150 ml), H ₂ O (300 ml) and H ₂ NOH HCl (55, 2, 0.80 mol) were dissolved together and Na ₂ CO ₃ (55.2 g, 0.53 mol) was added in small portions. The mixture was stirred at 23 ° C for 14 h, most of the THF was evaporated in vacuo, adjusted to pH> 10 with 50% aqueous NaOH, extracted with EtOAc (5 x 50 ml) ) and concentrated to a white foam. It was powdered with hexane, filtered and the solid dried in vacuo to give 40.31 g of the title compound.

The above residue was dissolved in EtOH (absolute, 1 L) and pre-nickel (50 ml suspension in EtOH) was added and the mixture was reduced under an atmosphere of H ₂ (50 psi = 50x6,895 kN / m ² ) for 3.5 h. The catalyst was filtered off and washed with EtOH. Concentration afforded 38.44 g (96% total) of the title compound as a colorless oil which solidified at -20 ° C.

c) 4-fluorophenyl-tolylsulfonomethylformamide

To a suspension of sodium p-toluenesulfinate (30 g) in water (100 ml) was added methyl tert-butyl ether (50 ml) and then conc. HCl (15 ml). After stirring for 5 min, the organic phase was removed and the aqueous phase was extracted with methyl tert-butyl ether. The organic phase was dried (Na ₂ SO ₄ ) and concentrated almost to dryness. Hexane was added and the resulting precipitate was collected to give p-toluenesulfinic acid: yield 22 g.

Combine p-toluenesulfonic acid <22 g, 140.6 mmol), p-fluorobenzaldehyde (22 ml, 206 mmol), formaldehyde (20 ml, 503 mmol) and camphorsulfonic acid (4 g, 17.3 mmol) and mix. at 60 ° C for 18 h. The resulting solid was crushed and stirred with a mixture of MeOH (35 ml) and hexane (82 ml) and then filtered. The solid was resuspended in MeOH / hexane (1: 3, 200 ml) and stirred vigorously to break the remaining large pieces. Filtration gave the title compound (27 g, 62% yield): Ή-NMR <400 MHz CDCl ₃ ): d 8.13 (s, 1H), 7.71 (d, 2H), 7.43 (dd, 2H) , 7.32 (d, 2H), 7.08 (t, 2H), 6.34 (d, 1H), 2.45 (s, 3H).

f) 4-fluorophenyl-tolylsulfonomethylisoyanide

4-Fluorophenyl-tolylsulfonomethylformamide (2.01 g, 6.25 mmol) in DME (32 ml) was cooled to -10 ° C, POCl ₃ (1.52 ml,

16.3 mmol) and then dropwise triethylamine (4.6 ml, 32.6 mmol) in DME (3 ml) while maintaining the temperature below -5 ° C. The mixture was gradually warmed to room temperature over a period of 1 hour, poured into water and extracted with EtOAc. The organic phase was washed with saturated aqueous NaHCO ₃ solution, dried (Na ₂ SO ₄ ) and concentrated. The residue was triturated with petroleum ether and filtered to give the title compound (1.7 g, 90% yield): Ή-NMR (CDCl ₃ ): d 7.63 (d, 2H), 7.33 ( m, 4H), 7.10 (t, 2H), 5.60 (s, ΙΗ), 2.50 (s, 3H).

g) 2-methoxypyrimidine-4-carboxaldehyde [1-tert. -butoxycarbonyl-4-aminopiperidine] imine

The product of Example 1 (d) (0.308 g, 2.23 mmol) and the product of Example 1 (c) (0.468 g, 2.34 mmol) were combined in CH ₂ Cl ₃ (50 ml) and stirred at 23 ° C for 16 h. Concentration gave the title compound as a light orange foam. Ή-NMR (CDCl ₃ ): d 8.56 (d, 1), 8.26 (s, 1), 7.57 (d, 1), 4.05 (s and m, 4), 3.5 (m, 2), 3.0 (m, 2), 1.75 (m, 4), 1.46 (s, 9).

h) 5- (2-methoxy-4-pyrimidinyl) -4- (4fluorophenyl) -1 - [(1-tert-butoxycarbonyl) -4piperidinyl] imidazole

The product of the above example, DMF (5 ml), the product of example 1 (f) (0.708 g, 2.23 mmol) and KjCO3 (0.308 g, 2.23 mmol) were mixed and stirred for 2 days, diluted with Et ₂ O and filtered. The filtrate was concentrated under high vacuum to give a brown solid. Trituration with Et ₂ O and hexane (1: 1, 200 ml) gave the title compound as a tan solid. Crystallization from acetone / hexane gave 0.505 g (64% of 5- (2-methoxy-4-pyridinyl) -4- (4-fluorophenyl) -1- (1-tert-butoxycarbonylpiperidin-4-yl) imidazole) . ESP + (Mass Spec) m / z 453 (MH ⁺ ).

i) 5- (2-methoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

The product of the previous example (0.505 g, 1.43 mmol) was added to ice-cooled TFA under argon. The resulting solution was warmed to 23 ° C and stirred for 1.5 h. The TFA was removed in vacuo, the residue was dissolved in EtOAc and extracted with water (2 x 20 ml). The combined aqueous phases were separated with EtOAc and cooled to 4 ° C, alkylated by the addition of 10% aqueous NaOH solution and the aqueous phase extracted with EtOAc (4 x 25 ml). The combined extracts were dried (Na ₂ SO ₄ ) and concentrated to give a white crystalline solid. Treatment of the solid with hexane gave 165 mg of a white solid. Evaporation of the above filtrate gave an additional 133 mg of light yellow crystals. Total yield (298 mg (59%)) For the first amount of crystals collected: mp 159-160 ° C.

Example 2.

5- (2-isopropoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

a) 2-methylthiopyrimidine-4-carboxaldehyde

The product of Example 1 (a) (9.96 g, 50 mmol) and 3N HCl (42 ml, 126 mmol) were combined and stirred at 48 ° C for 16 h. the mixture was cooled to 23 ° C, combined with EtOAc (200 ml) and basified by the addition of solid Na ₂ CO ₃ (12.6 g, 150 mmol). The aqueous phase was extracted with EtOAc (4x150 ml), dried (1 ₂ SO ₄ ), concentrated and the residue filtered through a layer of silica (150 ml) with CH ₂ Cl ₂ to give 7.49 g (97%) ) from the title compound. Ή-NMR (CDCl ₃ ): 59.96 (s, 1), 8.77 (d, 1), 7.44 (d, 1), 2.62 (s. 3).

b) 2-methylthiopyrimidine-4-carboxaldehyde-1-tert-butoxycarbonyl-4-aminopiperidine imine.

The product from the previous step (4.84 g,

31.4 mmol), MgSO ₄ (are 2 g), the product of Example 1 (d) (6.51 g, 32.6 mmol) and CH ₂ Cl ₂ (100 ml) were combined and stirred at 23 ° C for 23 h. 16 h. Filtration and concentration of the filtrate afforded the title compound as a yellow oil. ^: Η-NMR (CDC1 _3): 8,57 (d, 1), 8,27 (s. 1), 7.58 (d, 1), 4,05 (m, 2), 3,55 (m, 1 ), 3.00 (m. 2), 2.60 (s, 3), 1.75 (m, 4), 1.48 (s, 9).

c) 5- (2-methylthio-4-pyrimidinyl) -4- (4fluorophenyl) -1 - [(1-tert-butoxycarbonyl) -4piperidinyl] imidazole

The product of the previous example and the product of example 1 (f) (9.41 g, 32.6 mmol), DMF (64 ml) and K ₂ CO ₃ (4.43 g, 32.4 mmol) were reacted by the procedure of example 1 (h) to give 9.07 g of product (62% of the product of Example 1 (a). MS ES + m / z = 470 (MH ⁺ ).

d) 5- (2-methylsulfinyl-4-pyrimidinyl)

4- (4-fluorophenyl) -1 - ((1-tert-butoxycarbonyl) -4-piperidinyl] -imidazole.

The product of the previous example (4.69 g, 10 mmol) was dissolved in THF, cooled to -10 ° C and the oxonium salt (6.14 g, 10 mmol) in water (50 ml) was added dropwise (T < 5 ° C). The resulting mixture was warmed to 20 ° C for 50 min, poured into a vigorously stirred mixture of 10% aqueous NaOH solution (300 ml), ice (100 ml) and EtOAc (300 ml). The EtOAc layer was separated, dried (Na ₂ SO ₄ ) and concentrated to a yellow oil. Flash chromatography (0-2% MeOH in CH ₂ Cl ₂ ) gave 3.58 g (74%). ESP + (Mass Spec) m / z 486 (MH ⁺ ).

e) 5- (2-Isopropoxy-4-pyrimidinyl) -4 (4-fluorophenyl) -1 - [(1-tert-butoxycarbonyl) 4-piperidinyl] imidazole

With THF, it was washed with NaH (60% in mineral oil) and separated with more THF (5 ml) and anhydrous isopropanol (1.15 ml) was added. When boiling subsides, the resulting solution was re-cooled to 23 ° C and the product of the previous example (0.58 g, 1.19 mmol) in THF (5 ml) was added dropwise. After 5 min, the reaction mixture was shaken with EtOAc (100 ml) and water (50 ml) and the phases separated, the ethyl acetate layer dried and concentrated. The residue was crystallized from acetone / hexane to give 335 mg of the title compound (58%). MS ES + m / z = 482 (MH ^{&lt; + &gt;} ).

f) 5- (2-Isopropoxy-4-pyrimidinyl) -4 (4-fluorophenyl) -1- (4-piperidinyl) imidazole

The product of the previous example (325 mg, 0.68 mmol) was reacted with TFA according to the procedure of Example 1 (i). The crude product was crystallized from Et ₂ O / hexaH to give 106 mg (41%) of white crystals. M.P. 121-122 ° C.

Example 3.

5- (2-hydroxy-4-pyrimidinyl) -4- (4fluorophenyl) -1- (4-piperidinyl) imidazole trifluoroacetate

a) 5- (2-methylsulfonyl-4-pyrimidinyl) 4- (4-fluorophenyl) -1 - ((1-tert-butoxycarbonyl) -4-piperidinyl] imidazole.

The product of Example 2 (c) (9.07 g, 19.3 mmol) dissolved in THF was cooled to -10 ° C and the oxonium salt (28.5 g,

46.4 mmol) in water (250 ml). The resulting mixture was stirred at 23 ° C for 24 h, mixed with ice (100 ml) and CH ₂ Cl ₂ and washed with aqueous sodium chloride (100 ml), dried (Na ₂ SO ₄ ). and concentrated in vacuo to give 8.27 g (85%), MS ES + m / z = 502 (MH ⁺ ).

b) 5- (2-hydroxy-4-pyrimidinyl) -4- (4fluorophenyl) -1 - ((1-tert-butoxycarbonyl) -4piperidinyl] imidazole

The product of the previous example (141 mg, 0.28 mmol) was dissolved in THF (5 ml) and to this solution was added a 50% aqueous solution of NaOH (150 µL, 1.8 mmol). The solution was stirred for 3 days and a precipitate formed. The solid was filtered off, washed with THF, and dried in vacuo to give the title compound. MS ES + m / z = 440 (MH ^{&lt; + &gt;} ).

c) 5- (2-hydroxy-4-pyrimidinyl) -4- (4fluorophenyl) -1- (4-piperidinyl) imidazole

The product of the previous example and TFA (3 ml) was combined and stirred for 30 min, concentrated and the residue triturated with Et ₂ O and filtered, the white solid washed with Et, O, dried in vacuo to give 114 mg (90% mono-TFA salt of the product of Example 3 (a), mp 80-110 ° C (decomposition).

Example 4.

5- (2-methylthio-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

The product of Example 2 (c) is reacted with the product of Example 1 (i) to give the title compound, m.p. 182183 ° C.

Example 5. 5- (2-Methylthio-4-pyrimidinyl) -4- (4-fluorophenyl) -1 - ((1-methyl) -4-piperidinyl] imidazole

a) 2-methylthiopyrimidine-4-carboxaldehyde-1-methyl-4-aminopiperidinimine

The product of Example 2 (a) was reacted with 1-methyl 4-aminopiperidine according to the procedure of Example 2 (b) to give the title compound.

b) 5- (2-methylthio-4-pyrimidinyl) -4- (4fluorophenyl) -1 - ((1-methyl) -4-piperidinyl] imidazole

The product of the preceding example is reacted with the product of example 1 (f) according to the procedure of example 1 (i) to give the title compound. M.P. 181-182 ° C.

Example 6

5- (2-ethoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

a) 5- (2-Ethoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1 - ((1-tert-butoxycarbonyl) -4-piperidinyl] imidazole

The title compound was prepared by the method of Example 2 (e), except that anhydrous EtOH was used instead of 2-propanol.

B) 5- (2-Ethoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole

The product of the previous example was treated with TFA according to the procedure of Example 1 (ί) to give the title compound as white crystals. M.P. 128129 ° C.

Example 7.

- (1-ethoxycarboxylpiperidin-4-yl) -3 (4-thiomethylphenyl) -5- [2- (thiomethyl) pyrimidin4-yl] imidazole

a) 4-Thiomethylphenyl-tolylsulfonomethylisocyanide

The title compound was prepared according to the procedure of Example 1 (e) and 1 (f) using 4-thiomethylbenzaldehyde instead of 4-fluorobenzaldehyde.

b) 2-Thiomethylpyrimidine-4-carboxaldehyde- [1-ethoxycarbonyl-4-aminopiperidine] imine

The title compound was prepared using the procedure of Example 2 (b), but instead of the 1-tert-butoxycarbonyl4-aminopiperidine, the commercial product 1-ethoxycarbonyl-4-aminopiperidine was used.

c) 1- (1-ethylcarboxylpiperidin-4-yl) -3 (4-thiomethylphenyl) -5- [2- (thiomethyl) pyrimidin4-yl] imidazole

According to the procedure of Example 1 (h), 4-thiomethylphenyl-tolylsulfonomethylisocyanide (9.0 g, 29.2 mmol) and 2-thiomethylpyrimidine-4-carboxaldehyde- [1-ethoxycarbonyl-4-aminopiperidine] imine (7.0 g, were reacted). 22.1 mmol). After completion of the reaction, most of the DMF was evaporated under high vacuum, the remaining solution was poured into water and extracted with EtOAc. The extracts were washed with water, aqueous sodium chloride, dried (Na ₂ SO ₄ ), filtered, concentrated and flash chromatographed (60% EtOAc / hexane to give the title compound (4.0 g, 38, 5% yield) ESP + (Mass Spec) m / z 471 (MH ⁺ ).

Example 8.

1- (1-ethylcarbonylpiperidin-4-yl) -4- (4methylsulfinylphenyl) -5- [2-methylsulfinylpyrimidin-4-yl] imidazole

The product of the previous example (2 g,

4.26 mmol) was dissolved in THF, cooled to -10 ° C and oxone (3.3 g, 8.53 mmol) in water (10 ml) (T <5 ° C) was added dropwise. The resulting mixture was warmed to 20 ° C over a period of 50 min, poured into a vigorously stirred mixture of 10% aqueous NaOH solution (150 ml), ice (100 ml) and EtOAc separated, dried (Na ₂ SO ₄ ). and concentrated to give a yellow solid. Recrystallization from EtOAc / hexane (1:10) gave the title compound (80 mg). ESP + (Mass Spec) m / z 502 (MH ⁺ ).

All publications including, but not limited to, patents and patent applications cited in this specification are incorporated herein by reference.

The description fully discloses the invention, including its preferred embodiments. Modifications and improvements to the described embodiments are within the scope of the following claims. Without further elaboration, it is possible to apply the invention to its full extent. The examples illustrate but do not limit the scope of the present invention.

Claims

Claims (25)

Compounds of formula wherein R 1 represents a 4-pyrimidinyl ring substituted with C ₁₄ alkoxy and optionally optionally substituted independently by C ₁₄ alkyl, halogen, hydroxy, C _1-4 alkoxy, C ₄ alkylthio, C _{| 4} alkylsulfinyl, CH ₂ OR ₁₂ , amino, mono and di-C ₁₆ alkyl substituted amino, N (R ₁₀ ) C (O) R ₍ . Or N-heterocyclyl ring having a ring having from 5 to 7 atoms and optionally contains an additional heteroatom selected from oxygen, sulfur or NR ₁₅ ; R ₄ denotes phenyl, naphth-1-yl or naphth-2-yl, or heteroaryl, which is optionally substituted by one or two substituents, each of which is independently selected and which substituent on 4-phenyl, 4-naphth-1-yl, 5-naphth-2-yl or 6-naphth-2-yl is halogen, cyano, nitro, C (Z) NR ₇ R ₁₇ , C ( Z) OR ₁₆ , iCR ₁₀ R ₂₀ ) _v COR ₁₂ , SR ₅ , SORj, OR ₁₂ substituted with halogen C _1-4 alkyl, C _1-4 alkyl, ZC (Z) R ₁₂ , NR ₁₀ C (Z) R ₁₆ or _{(CR 1O R 2O) V NR} 1O R, O and which, for other positions of substitution is halogen, cyano, C (Z) NR ₁₃ R _14, C (Z) OR _3, (CR. ₀ R ₂₀ ) _m „COR ₃ , S (O) _m R ₃ , OR ₃ substituted with halogen C _b4 alkyl, C ₁₄ alkyl, (CR ₁₀ R ₂₀ ) _m , .NR ₁₀ C (Z) R ₃ , NR ₁₀ S (O) _m , R ₈ , NR ₁₀ S (O) _ra . NR ₇ R ₁₇ , ZC (Z) R ₃ or (CR ₁₀ R ₂₀ ) _ra NR ₁₃ R ₁₄ ; n has a value from 1 to 10; m is 0, 1 or 2; m 'is 1 or 2; m 'is 0 or 1 to 5; v is 0 or 1 or 2; R ₂ is optionally substituted heterocyclyl or optionally substituted heterocyclyl C _1-10 alkyl moiety; Z is oxygen or sulfur; R _c is hydrogen, C ₁₆ alkyl, C ₃ . ₇ cycloalkyl, aryl, aryl C _1-4 alkyl, heteroaryl, heteroaryl C ₁₄ alkyl, heterocyclyl or heterocyclyl C ₁₄ alkyl; R ₃ is heterocyclyl, heterocyclyl C, _ ₁₀ allyl or R _8; R ₃ is hydrogen, C ₁₄ alkyl, C _2-4 alkenyl, C _2-4 alkynyl, or NR ₇ R ₁₇ , except where -SR ₅ is -SNR ₇ R ₁₇ and -SOR 1 is -SOH; R ₇ and R ₁₇ are independently selected from hydrogen or C ₁₄ alkyl or R ₇ and R ₁₇ together with the nitrogen to which they are attached form a heterocyclic ring of 5 to 7 atoms, which ring optionally contains an additional heteroatom - oxygen, sulfur or NR _I? ; R _s is C,. ₁₀ alkyl substituted with halogen C ₁₁₀ alkyl, C ₂₁₀ alkenyl, C ₂₁₀ alkynyl, C _3-7 cycloalkyl, C _5-7 cycloalkenyl, aryl, aryl C ₁₁₀ alkyl, heteroaryl, heteroaryl alkyl, (CR ^ R ^ OR ,,, ( CR ₁₀ R ₂₀ ) _n S (O) _m R ₁₈ , (CR ₁₀ R ₂₀ ) _n NHS (O) ₂ R, ₈ , (CR ₁₀ R ₂₀ ) _n NR ₁₃ R ₁₄ , such as aryl, arylalkyl, heteroaryl, heteroarylalkyl may be optionally substituted; R 1 is hydrogen, C (Z) R _H, or optionally substituted C ₁₁₀ alkyl, S (O) ₂ R ₁₈ , optionally substituted aryl, or optionally substituted aryl C, _ ₄ alkyl; R ₁₀ and R ₂₀ are each independently selected from hydrogen or C _1-4 alkyl; R _H is hydrogen, C ,. _1o alkyl, _{C3 7} cycloalkyl, heterocyclyl, heterocyclic otsiklil C alkyl, aryl, aryl C _{1 -C 10} alkyl, heteroaryl or heteroaryl Cj _y alkyl; R ₁₂ is hydrogen or R _16; R ₁₃ and R ₁₄ are all independently selected from hydrogen or an optionally substituted C _1-4 alkyl, optionally substituted aryl, or optionally substituted aryl C ₁₄ alkyl, or together with the nitrogen atom to which they are attached, form a heterocyclic ring of 5 to 7 atoms, which ring optionally contains an additional heteroatom selected from oxygen, sulfur or NR, ; R ₁₃ is R _{1 ()} or C (Z) -C _1-4 alkyl; R ₁₆ is C _1-4 alkyl substituted with halogen C _1-4 alkyl or C ₃₇ cycloalkyl; R _{| g} is C, _{| 0} alkyl, C _{3 7} cycloalkyl, aryl, arylalkyl, heterocyclyl, heterocyclyl C ₁₁₀ alkyl, heteroaryl or heteroaryl alkyl; or pharmaceutically acceptable salts thereof. A compound according to claim 1, wherein R is substituted by isopropoxy, ethoxy or methoxy. A compound according to claim 1, wherein R ₄ is optionally substituted phenyl. Compound according to claim 3, characterized in that the phenyl is mono or polysubstituted independently with halogen, SR ₃ , S (O) R ₃ , OR ₁₂ substituted with halogen C _1-4 alkyl or C 1-6 alkyl. 1-4 5. A compound according to claim 4 wherein the phenyl is fluorine substituted. A compound according to claim 1 wherein R ₂ is morpholinopropyl, piperidine, N-methylpiperidine, Nbenzylpiperidine or 2,2,6,6-tetramethylpiperidine. The compound of claim 1, which is 1- (4-piperidinyl) -4- (4-fluorophenyl) -5- (2-isopropoxy-4-pyrimidinyl) imidazole or a pharmaceutically acceptable salt thereof. A compound according to claim 1, which is 1- (4-piperidinyl) -4- (4-fluorophenyl) -5- (2-methoxy-4-pyrimidinyl) imidazole or a pharmaceutically acceptable salt thereof. A compound according to claim 1 comprising 5- (2-ethoxy-4-pyrimidinyl) 4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier or diluent. A method for treating mammalian inflammation, wherein the mammal in need of treatment is treated with an effective amount of a compound of formula I according to claims 1 to 9. A method of treating mammals for disease with a mediator CSBP / RK / p38 kinase, characterized in that the mammal in need is treated with an effective amount of a compound of formula I according to claims 1 to 9. 13. The method of claim 12, wherein the disease mediated CSBP / RK / p38 kinase is psoriasis arthritis, Reiter syndrome, rheumatoid arthritis, gout, gouty arthritis, traumatic arthritis, rubella arthritis, and acute synovitis, rheumatoid arthritis arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis, and other arthritic conditions, sepsis, septic shock, endotoxic shock, gram-negative sepsis, toxic shock syndrome, stroke, neurotrauma, respiratory malignancy, distress, cerebral distress syndrome, ablation, silicosis, pulmonary sarcoisosis, bone resorption disease, osteoporosis, restenosis, cardiac or renal reperfusion injury, thrombosis, glomerulonephritis, diabetes, transplant response to the recipient, allograft rejection, colitis, collagen, dermatitis, psoriasis, sunburn and conjunctivitis. A process for the preparation of a compound of formula I as defined in claim 1, wherein the compound of formula A-S (O) _p ^r 4 NC is reacted with a compound of formula Rt nr ₂ V III Η where ρ is 0 or 2, and a base strong enough to deprotonate the isonitrile residue of formula II, R _p R ₂ and R ₄ are defined in claim 1 or are precursors of the groups R 1, R ₂ , R ₄ and Ar is in optionally substituted phenyl group and then, if necessary, the precursors of the groups Rp R ₂ and R ₄ are converted to the groups R ₍ , R ₂ and R ₄ . 15. The method of claim 14 wherein p is 0. 16. The method of claim 14, wherein p is 2. A process according to claim 16, wherein the base is an amine, carbonate, hydride or alkyl- or aryl lithium reagent. A method according to claim 16, characterized in that the imine of formula III is isolated before reacting with the compound of formula II. A method according to claim 16, characterized in that the imine of formula III is prepared in situ prior to reaction with the compound of formula II. 20. The method of claim 19, wherein the imine is prepared in situ by the reaction of an aldehyde of formula R ^ HO in which R 1 has the meanings defined for formula I with a primary amine of formula R ₂ NH ₂ in which R ₂ has the meanings defined for Formula I. 21. The method of claim 20, wherein dehydration conditions are used to form imine in situ. 22. The method according to claim 16, 20 or 21, wherein the solvent is N, N-dimethylformamide (DMF), halogenated solvent, tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), alcohol, benzene, toluene, MeCN or DME . 23. A method according to claim 20, characterized in that the aldehyde R ^ HO is pyrimidine aldehyde of the formula wherein X is C _1-4 alkoxy and X, is defined as the optional substituent on the residue R _I in formula I according to claim 1, to obtain a compound of formula I or a pharmaceutically acceptable salt thereof. A method according to claim 20, wherein the R ₂ residue in the primary amine R ₂ NH ₂ is piperidine, 1-formyl-4-piperidine, 1-benzyl-4-piperidine, 1-methyl-4-piperidine, 1-ethoxycarbonyl-4-piperidine, 1-tert-butoxycarbonyl-4-piperidine, 2,2,6,6-tetramethyl-4-piperidine, morpholinoethyl, morpholinopropyl, pyrrolidinylpropyl or piperidinylpropyl. A method according to claims 14 to 24, characterized in that the compound is: 1- (4-piperidinyl) -4- (4-fluorophenyl) -5 (2-isopropoxy-4-pyrimidinyl) imidazole, 1- (4-piperidinyl) -4- (4-fluorophenyl) -5 (2-methoxy-4-pyrimidinyl) imidazole or 5- (2-Ethoxy-4-pyrimidinyl) -4- (4-fluorophenyl) -1- (4-piperidinyl) imidazole or any of their pharmaceutically acceptable salts.