BG107013A - Il-8 receptor antagonists - Google Patents

Abstract

This invention relates to novel compounds of formula and compositions thereof, useful in the treatment of diseases states mediated by chemokine, Interleukin-8 (IL-8). 13 claims

Description

Technical field

The invention relates to novel sulfonamide substituted diphenyl ureas, methods for their preparation and use for the preparation of pharmaceutical compositions for the treatment of diseases mediated by IL-8, GROa, GROp, GROy, NAP-2 and ENA-78.

BACKGROUND OF THE INVENTION

Many different names have been used for interleukin-8 (IL8) such as non-neutrophil attractant / activating protein-1 (NAP-1), monocytically derivatized non-neutrophil chemotactic factor (MDNCF), non-neutrophil activating factor (NAF), and T-cell lymphocytic hematocytic factor. Interleukin-8 is a chemoattractant for neurophiles, basophils and a subset of T cells. It is produced by a variety of nucleated cells, including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-1α, IL-Ιβ, or LPS, and nucleophiles themselves when exposed to LPS or chemotactic factors such as FMLP. M. Baggiolini et al., J. Clin. Invest. 84, 1045 (1989); J. Schroder et al., J. Immunol. 139, 3474 (1987) and J. Immunol. 144, 2223 (1990); Strieter, et al., Science 243, 1467 (1989) and J. Biol. Chem., 264,10621 (1989); Cassatella et al., J. Immunol. 148, 3216 (1992).

GROa, GROp, GROy and NAP-2 also belong to the chemokine family. Like IL-8, these chemokines are also referred to as different

MMSM

names. For example, GROa, β, and γ are referred to as MGSAa, β, and γ, respectively (Melanoma Growth Stimulating Activity), see Richmond et al., J. Cell Physiology 129, 375 (1986) and Chang et al., J. Immunol. 148, 451 (1992). All these chemokines of the α family that possess the ELR motif, directly preceding the CXC motif, bind to the IL-8 B receptor (CXCR2).

IL-8, GROa, ΰΒΟβ, GROy, NAP-2 and ENA-78 stimulate many functions in vitro. All of them have been shown to possess neutrophil chemoattractant properties, whereas IL-8 and GROa have demonstrated T-lymphocyte and basophilic chemotactic activity. In addition, IL-8 can induce histamine release from basophils from normal and atopic individuals. In addition, GROa and IL-8 can induce lysosomal enzymatic release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mas-1 (CD11b / CD18) on neutrophils without de novo protein synthesis. This may contribute to increased adhesion of neutrophils to vascular endothelial cells. Many known diseases are characterized by massive neutrophil infiltration. As IL-8, GROa, GROp, GROy and NAP-2 promote the accumulation and activation of neutrophils, these chemokines are involved in a wide range of acute and chronic inflammatory disorders, including psoriasis and rheumatoid arthritis, Baggiolini et al., FEBS Lett. 307, 97 (1992); Miller et al., Crit. Rev. Immunol. 12.17 (1992); Oppenheim et al., Annu. Rev. Immunol. 9, 617 (1991); Seitz et al., J. Clin. Invest., 87, 463 (1991); Miller et al., Am. Rev. Respir. Dis., 146, 427 (1992); Doonely et al., Lancet 341,643 (1993). In addition, ELR chemokines (those having the amino acid ELR motif immediately before the CXC motif) have also been implicated in angiostasis, Strieter et al., Science 258, 1798 (1992).

In vitro, IL-8, GROa, GROp, GROy, and NAP-2 induce neutrophil-induced alteration, chemotaxis, granular release, and respiratory burst by binding to and activating the seven-transembrane receptors, the G-protein coupled family, and in particular by binding to IL -8 receptors, especially the ΙΙ_-1β receptor (CXCR2), Thomas et al., J. Biol. Chem, 266,14839 (1991); and Holmes et al., Science 253, 1278 (1991). The development of non-peptide small molecule antagonists for members of this receptor family has precedent. For an overview, see R. Freidinger in: Progress in Drug Research, Vol. 40, p. 33 - 98. Birkhauser Verlag, Basel 1993. From here, the IL-8 receptor is a promising target for the development of new anti-inflammatory agents.

Two high-affinity human IL-8 receptors (77% homology) were characterized: IL-8Ra, which binds only high affinity IL-8, and IL-8RB, which has high affinity for IL-8 as well as GROa, GROp. GROy and NAP-2. See Holmes et al., Supra; Murphy et al., Science 253, 1280 (1991); Lee et al., J. Biol. Chem., 267, 16283 (1992); LaRosa et al., J. Biol. Chem., 267, 25402 (1992) and Gayle et al., J. Biol. Chem., 268, 7283 (1993).

There remains a need for treatment compounds in the art that are capable of binding to the IL-8α or β receptor. Therefore, conditions associated with increased production of IL-8 (which is responsible for chemotaxis and reduction of neutrophil T cells at the site of inflammation) will be favorably affected by compounds that are inhibitors of IL-8 receptor binding.

SUMMARY OF THE INVENTION i

The invention provides compounds of formula (I) for treating a chemokine mediated disease, wherein the chemokine is one that binds to the IL-8α or β receptor. In particular, the chemokine is IL-8.

The invention also relates to the use of compounds of general formula (I) for the preparation of pharmaceutical compositions for inhibiting the binding of IL-8 to its receptor in a mammal in need thereof.

The invention also provides novel compounds of formula (I) and pharmaceutical compositions comprising a compound of formula (I) and a pharmaceutical carrier or diluent.

Compounds of formula (I) that are applicable in the present invention are represented by the structure:

wherein:

R _b is independently selected from the group consisting of hydrogen, NR ₆ R ₇ ,

OH, OR _a , C 1-5 alkyl, aryl, arylC _1-4 alkyl, arylC ₂ . ₄ alkenyl; cycloalkyl, cycloalkyl C _1-5 alkyl, heteroaryl, heteroarylC 1-6 alkyl, heteroarylC ₂ . ₄ alkenyl, heterocyclyl, heterocyclyl _b4 alkyl, heterocyclylC _{2. 4} alkenyl, these groups may optionally be substituted on one to three times independently from the group consisting of halogen, nitro, halogenzamestenS ^ e alkyl, C _V4 alkyl, amino, mono or di- C _V4 alkyl substituted amine, OR _a, C ( O) R _a , NR _a C (O) OR _a , OC (O) NR ₆ R ₇ , hydroxy, NR ₉ C (O) R _a , S (O) _m .R _a , C (O) NR 6 R ₇₁ C (O) OH, C (O) OR _a , S (O) ₂ NR ₆ R ₇ , NHS (O) ₂ R _a or the two R _b substituents together form a 3-10 membered ring, optionally substituted and containing in addition to carbon , pspzhimMami

independently from 1 to 3 groups selected from NR _a , O, S, SO and SO ₂ and wherein the substituent may be unsaturated;

R _a represents an alkyl, aryl, aryl ^ g alkyl, heteroaryl, heteroarylC ^ g alkyl, heterocyclyl, COOR _13, heterocyclyl C _b4 alkyl,

all of these groups may be optionally substituted;

m is a number from 1 to 3;

rn 'is 0 or a number from 1 to 2;

η is a number from 1 to 3;

q is 0 or a number from 1 to 10;

t is 0 or a number from 1 to 2;

s is a number from 1 to 3;

Rf is independently hydrogen, halogen, nitro, cyano, R _V1O alkyl, halosubstituted R _b1 o alkyl, R _{2. 10} alkenyl, R ^ q а alkoxy, halogen substituted R- |. ₁₀ alkoxy, azide, S (O) _t R ₄ , (CR ₈ R ₈ ) _q S (O) _t R ₄ , hydroxy, hydroxy substituted C 1-6 alkyl, aryl, aryl C 1-4 alkyl, aryl R ₂ . ₁₀ alkenyl, aryloxy, arylC _1-4 alkyloxy, heteroaryl, heteroarylalkyl, heteroaryl R ₂ . ₁₀ alkenyl, heteroarilR _b4 alkyloxy, heterocyclyl, heterocyclyl _{L 4} alkyl, heterocyclyl _D4 alkyloxy, heterocyclyl _{2. 10} alkenyl, (CR ₈ R ₈ ) q NR ₄ R ₅ , (CR ₈ R ₈ ) _q C (O) NR ₄ R ₅ , R ₂ . ₁₀ anKeHHnC (O) NR ₄ R ₅ , (CR ₈ R ₈ ) qC (O) NR ₄ R ₁₀ , S (O) ₃ R ₈ , (CR ₈ R ₈ ) _q C (O) Ri ₁ , R ₂ . ₁₀ alkenylC (O) P ₁₁ , R ₂ . ₁₀ alkenylC (O) OB _1b (CRgR ^ CiOJORn, (CRgR ^ OCiCORn, (CRgR ^ qNR.CpjRn, (CR ₈ R ₈ ) _q C (NR ₄ ) NR ₄ R ₅ , (CRsR ^ qNR.CiNRgJR ^, ( CR ₈ R ₈ ) _q NHS (O) ₂ R ₁₃ and (CR ₈ R ₈ ) _q S (O) ₂ NR ₄ R ₅ or two R 4 groups together form a 0- (4-O- or 5 or 6 membered sat. or an unsaturated ring and the alkyl, aryl, aralkyl, heteroaryl, heterocyclyl groups may be optionally substituted;

R ₄ and R ₅ are independently hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylC ^ alkyl, optionally substituted heteroaryl, optionally substituted heteroarilC _V4 alkyl, heterocyclyl, heterocyclyl alkyl or R ₄ and R ₅ together of nitrogen same, «l

b to which they are attached form a 5 or 7 membered ring which optionally contains an additional heteroatom selected from oxygen, nitrogen or sulfur;

R ₆ and R ₇ are independently hydrogen, C _1-4 alkyl, heteroaryl, aryl, alkylaryl and alkylC _1-4 heteroalkyl or R ₆ and R ₇ together with the nitrogen to which they are attached form a 5 or 7 membered ring, optionally containing an additional heteroatom which is selected from oxygen, nitrogen or _; sulfur and the ring may be substituted;

Y is CR ₁₄ R _15, NR _14i 0, CO, and S (O) t; R ₈ is hydrogen or C _1-4 alkyl;

: φ R ₉ is C 1-6 alkyl;

R ₁₀ is C 1-6 alkyl C (O) ₂ R ₈ , t

R 11 is hydrogen, optionally substituted C 1-6 alkyl, optionally substituted aryl, optionally substituted arylC 1-5 alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC 1-5 alkyl, optionally substituted heterocyclyl, and optionally substituted heterocyclyl;

R ₁₂ is hydrogen, C _b4 alkyl, aryl, arylC ^ alkyl, heteroaryl, heteroarylC _{1. 4} alkyl, heterocyclyl and heterocyclylC 1-4 alkyl;

R ₁₃ is Smapkil, aryl, aryl C _m alkyl, heteroaryl, heteroarilC _V4 alkyl, heterocyclyl and heterocyclylC ^ alkyl;

u R ₁₄ and R ₁₅ are independently hydrogen, optionally substituted C _1h alkyl,

OR _a and NR ₄ R ₅ ; or R ₁₄ and R ₁₅ together with the atom (s) to which they are attached may form a 4 to 7 membered ring, which may optionally contain an additional heteroatom, which heteroatom is selected from the group oxygen, nitrogen and sulfur, wherein the ring may optionally be be substituted;

or a pharmaceutically acceptable salt thereof.

The compounds of formula (I) may also be used in veterinary practice in mammals other than humans in need of inhibition of IL-8 or other chemokines that bind to IL-8 α ί

ί ι

or β receptors. Chemokine mediated diseases that can be treated, treated or prophylactically, in animals include disease states such as those described in the Compound Use section.

Suitably, R _b is, independently, hydrogen, NR ₆ R _7, OH, OR _a, C _b4 alkyl, aryl, arylC _{1. 4} alkyl, arylC ₂ . ₄ alkenyl; heteroaryl, heteroarylC _b4 alkyl, heteroarylC _{2. 4} alkenyl, heterocyclyl, heterocyclylC _{1. 4} alkyl or heterocyclylC ₂ . ₄ alkenyl, all of these groups may optionally be substituted one to three times independently of halogen, nitro, halogen substituted Cl ₁ . ₄ alkyl, _b4 alkyl, amino, mono or di-C _b4 alkyl substituted amine, cycloalkyl, cycloalkyl C _{V4 alkyl, OR a, C (O)} R a, NR a C (O) OR a, OC (O) NR ₆ R ₇ , aryloxy, aryl C _{V 4} oxy, hydroxy, C 1-4 alkoxy, NR ₉ C (O) R _a , S (O) _m .R _a , C (O) NR ₆ R ₇ , C (O) OH, C (O) OR _a , S (O) ₂ NR ₆ R ₇ , NHS (O) ₂ R _a . Alternatively, the two R _b substituents may co-form a 3-10 membered ring optionally substituted and in addition to carbon independently from 1 to 3 groups, NR ₉ , O, S, SO and SO _2, and which may be optionally substituted.

Suitably, R _a can intended a alkyl, aryl, aryl C _m alkyl, heteroaryl, heteroarylC _b4 alkyl, heterocyclyl or heterocyclyl-alkyl, each of these radicals can be substituted.

Suitably, Rj is, independently, hydrogen, halogen, nitro, cyano, halosubstituted R _M0 alkyl such as CF _3, R _{1. 10} alkyl such as methyl, ethyl, isopropyl or n-propyl, R ₂ . ₁₀ alkenyl, R _H0 alkoxy such as methoxy or ethoxy; halogen substituted R ₁ . ₁₀ alkoxy such as trifluoromethoxy, azide, (CR ₈ R ₈ ) _q S (O) _t R ₄ where t is 0, 1 or 2, hydroxy, hydroxy C _1-4 alkyl such as methanol or ethanol, aryl as phenyl or naphthyl, aryl C 1-4 alkyl such as benzyl, aryloxy such as phenoxy, aryl C 1-6 alkyloxy such as benzyloxy, heteroaryl, heteroarylalkyl, heteroarylP 1-6 alkyloxy, aryl R ₂ . ₁₀ alkenyl, heteroaryl R ₂ . ₁₀ alkenyl, heterocyclylC ₂ . ₁₀ alkenyl, (CR ₈ R ₈ ) _q NR ₄ R ₅ , R ₂ . ₁₀ anKeHnnC (O) NR ₄ R ₅ , (CR ₈ R ₈ ) _q C (O) NR ₄ R ₅ , (CR ₈ Re) _q C (O) NR ₄ R ₁₀ , S (O) ₃ H, S ( O) ₃ Re, (0 ^ 0)%, R ₂ . ₁₀ alkenyl

CO,! And oalkenyl) f, (CR ^ cm), (CR ^ C ^ OR) ,, (CB ₈ B ₈ ) _q OC (O) B _1b (CB ₈ B ₈ ) _q NB ₄ C (O) R ₁₁ , (CR ₈ R ₈ ) _q C (NR ₄ ) NR ₄ R ₅ , (CR ₈ R ₈ ) _q NR ₄ C (NR ₅ ) R ₁₁ , (CR ₈ R ₈ ) _q NHS (O) ₂ R _13> (CR ₈ R ₈ ) _q S (O) ₂ NR ₄ R ₅ . All radicals containing aryl, heteroaryl and heterocyclyl may be optionally substituted as defined below.

The term "drill, heteroaryl, and heterocyclyl" moieties used herein refers to the ring and alkyl, or, if included, to alkenyl rings such as aryl, arylalkyl and aryl alkenyl rings. The terms "parts" and "rings" may be used interchangeably in the description.

Suitably, R ₄ and R ₅ are independently hydrogen, optionally substituted C ₁ . ₄ alkyl, optionally substituted aryl, optionally substituted aryl C _b4 alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C _m alkyl, heterocyclyl, heterocyclyl alkyl or R @ ₄ and R _& along with the nitrogen to which they are attached form a 5 to 7 membered ring optionally containing an additional heteroatom selected from oxygen, nitrogen or sulfur;

Suitably R ₆ and R ₇ are independently hydrogen, C _1-4 alkyl, heteroaryl, alkylC 1-6 heteroalkyl or R ₆ and R ₇ together with the nitrogen to which they are attached form a 5 or 7 membered ring, optionally containing an additional heteroatom, which is selected between oxygen, nitrogen or sulfur and the ring may be optionally substituted;

Suitably, R ₈ is independently hydrogen or C _1-4 alkyl;

Suitably, R ₉ is hydrogen or C _1-4 alkyl;

Suitably q is 0 or an integer from 1 to 10;

Suitably R ₁₀ is C 1-6 alkyl C (O) ₂ R ₈ as CH ₂ C (O) ₂ H or

CH ₂ C (O) ₂ CH ₃ .

Suitably R _n is hydrogen, C _1h alkyl, aryl, aryl C _m alkyl, heteroaryl, heteroarylC ^ alkyl, heterocyclyl or heterocyclyl _m alkyl;

ί «<*

Suitably H ₁₂ is hydrogen, C 1-6 alkyl, aryl or arylalkyl.

Suitably, R ₁₃ is C _1-4 alkyl, aryl, arylalkyl, heteroaryl, heteroarylC 1-6 alkyl, heterocyclyl and heterocyclylC 1-6 alkyl wherein all moieties containing aryl, heteroaryl and heterocyclyl may be optionally substituted.

R ₁₄ and R ₁₅ are suitably hydrogen, optionally substituted C _b4 alkyl, OR _a, NR ₄ R _5, or R ₁₄ and R ₁₅ together with the atom (s) to which they are attached may form a 4 to 7 membered ring which may optionally contain an additional heteroatom, which heteroatom is selected from the group oxygen, nitrogen and sulfur, wherein the ring may be optionally substituted;

Suitably Y is CR ₁₄ R ₁₅ , NR ₁₄ , 0. * «H»

Suitably, R _a represents alkyl, aryl _b4 alkyl, heteroaryl, E heteroarylC ^ alkyl, heterocyclyl or heterocyclyl-alkyl, katof · «all these radicals may have been replaced. *

As used herein .eventualno substituted ", unless otherwise specified, shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine, hydroxy, hydroxy substituted C _G10 alkyl, _b10 alkoxy such as methoxy or ethoxy, 8 (0) ^ 0 C 1-6 alkyl, where m 'denotes 0, 1 or 2 such as methyl thio, methylsulfinyl or methylsulfonyl; amino, mono and disubstituted amino groups such as the NR ₈ R ₁₂ group, NHC (O) R ₁₃ , C (O) NR ₈ R ₁₂ , C (O) OH, S (O) ₂ NR ₈ R ₁₂ , NHS (O) ₂ R ₁₃ , C 1-6 alkyl, such as methyl, ethyl, propyl, isopropyl or t-butyl, halogen-substituted C 1-6 alkyl such as CF ₃ , optionally substituted aryl such as phenyl, or optionally substituted arylalkyl, such as benzyl or phenethyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, wherein these aryl, heteroaryl or heterocyclyl groups may be substituted once or twice with halogen; hydroxy; hydroxy substituted alkyl; C 1-6 alkoxy; S (O) _m -C _M0 alkyl, amino, mono and *

I disubstituted amino group as NR ₆ R ₇ group; C 1-4 alkyl or hapogen-substituted C 1-6 alkyl such as CF ₃ .

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, hydrobromic, sulfuric, phosphoric, methanosulfonic, ethanosulfonic, acetic, malic, tartaric, citric, lactic, oleic, lactic, oleic , benzoene, >>

salicylic, phenylacetic and almond acid. In addition, pharmaceutically acceptable salts of the compounds of formula (I) may be prepared with a pharmaceutically acceptable cation. Suitable

Pharmaceutically acceptable cations are well known in the art and include alkali, alkaline earth, ammonium and quaternary ammonium cations.

The following terms used mean:

.Hapogen refers to all halogen elements, ie. chlorine,>

fluorine, bromine and iodine.

'C _10-10 alkyl or' alkyl '- means straight or branched groups: a chain containing 1 to 10 carbon atoms, unless the chain is'>

j 1 'otherwise limited, including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl and the like .

"cycloalkyl" means a cyclic group, preferably containing 3 to 8 carbons, including, but not limited to, cyclopropyl, cyclopentyl, cyclohexyl and the like.

"alkenyl" means a straight or branched chain group: ι containing from 2 to 10 carbon atoms, unless the chain is otherwise limited, including, but not limited to, 1-propenyl, 2-propenyl, 2-methyl-1- propenyl, 1-bugenyl, 2-bugenyl and the like.

"Aryl" - phenyl and naphthyl;

"heteroaryl" (alone or in combination such as .heteroaryloxy or heteroarylalkyl) is a 5-10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O, or S such as, but not limited to, pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, tetrazole, thiazole, thiadiazole, triazole, imidazole or benzimidazole.

heterocyclyl (alone or in combination as. heterocyclyl alkyl) is a saturated or partially unsaturated 4-10 membered ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O or S as , but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, thiomorpholine or imidazolidine. Moreover, sulfur may be possibly oxidized to sulfone or sulfoxide.

.arilalkil "or .heteroarilalkil" represents _b10 alkyl as defined above, attached to an aryl, heteroaryl or heterocyclyl group, as defined above, unless otherwise indicated.

"ruffinyl" - is the oxide S (O) of the corresponding sulfide, the term "thio" refers to a sulfide, and the term "ruulfonyl" refers to a fully oxidized S (O) ₂ group.

"When two groups can together form a 5 or 6 membered saturated or unsaturated ring" means the formation of an aromatic ring system such as naphthalene or a phenyl group to which a 6 membered partially saturated or unsaturated ring such as C ₆ cycloalkenyl is attached; e. hexene or a C ₅ cycloalkenyl group such as cyclopentane.

Representative compounds of formula (I) include:

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 H -cyclohexylurea;

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 H -adamantyl) urea;

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N '- (tetrahydro-2-pyranyl) urea;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '- (3-tetrahydrofuryl) urea;

6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) ureido] benzenesulfonamide;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -1H-cyclohexylurea; 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethyl-cyclopropyl) ureido] benzenesulfonamide;

6-chloro-2-hydroxy-3- (3-piperidin-4-ylureido) benzenesulfonamide;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '- (4-methylcyclohexyl) urea;

6-chloro-2-hydroxy-3- [3- (3-methoxy-cyclohexyl) ureido] benzenesulfonamide;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N'-cyclopentylurea;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N'-cyclobutylurea;

N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N'-cyclopropylurea;

N- [6-Chloro-3- (3-cyclopentyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid tert-butyl ester;

- [4-chloro-2-hydroxy-3- (piperazine-1-sulfonyl) -phenyl] -3-cyclopentyl-urea;

N- [6-Chloro-3- (3-cyclobutyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid tert-butyl ester;

3- [3 - ((1b, 2S) -2-benzyloxy-cyclohexyl) -ureido] -6-chloro-2-hydroxy-benzenesulfonamide and

6-chloro-3- (3-cyclobugyl-ureido) -2-hydroxy-N '- dimethylbenzenesulfonamide.

Methods of preparation

The compounds of formula (I) can be prepared by use

of synthetic methods, some of which are illustrated in the schemes below. The syntheses given in these schemes are applicable to the preparation of compounds of formula (I) having different R, R _b and Y groups that interact using any substituents that are suitably protected to carry out the reactions described herein. In these cases, after deprotection, the described compounds are obtained. Once urea nuclei have been created, other compounds of these formulas can be prepared by using standard functional group conversion techniques well known to those skilled in the art.

Scheme 1

S0 ₉ NR'R

SO ₂ NR'R

0 '

SOoNR'R

and:

O '

f —►

a) I) NCS, AcOH, H ₂ O, II) NR'R 'H, pyr

b) H ₂ SO ₄ , HNO ₃ ; c) NaOAc, 18-crown-6

d) H ₂ SO ₄₁ MeOH e) Pd / C, H ₂ ; f) RCNO, DMF;

The desired 4-chloro N- (3-sulfonamido-2-hydroxy phenyl) -N 'cycloalkyl urea can be synthesized from commercially available 2,5-dichlorothiophenol using the method developed above in Scheme 1. The thiol can be oxidized to the corresponding sulfonyl halide using a halogenating agent such as NCS, NBS, Cl ₂ or Br ₂ in the presence of a protic solvent such as water, acetic acid or alcohol or a combination thereof. Yield may be increased if a buffering agent such as sodium or potassium acetate is incorporated into the reaction mixture and the reaction is carried out with an amine in the presence of a base such as pyridine, triethylamine, potassium carbonate or sodium hydride to form analogous sulfonamide 2 - Scheme 1.

The dichlorosulfonamide of Scheme 2 can be nitrated using strong nitrating conditions such as nitric acid in sulfuric acid to form aromatic nitro compound 3 of Scheme 1. The chlorine atom at ortho-position with respect to the nitro group can be selectively

hydrolyzed using an acetate salt such as sodium acetate in the presence of a crown ether such as 18-crown-6 to form acetate 4 of Scheme 1, the acetate group can be hydrolyzed under acidic conditions in an alcoholic solvent such as methanol or ethanol with catalytic amount of acid to form phenol-5. The nitro group can be reduced under well-known conditions such as hydrogen and palladium on

carbon, tin chloride in methanol, zinc in acetic acid or thiol to form the desired urea. Alternatively, the desired isocyanates may be obtained by condensation of an amine with triphosgene in the presence of a base (such as potassium carbonate) or by reaction of a carboxylic acid with diphenyl phosphoryl azide in the presence of a base (such as triethylamine).

Scheme 2

^{: 0}

0 '

a) NaH, R'X b) NaH RX

If sulfonamide-1 from Scheme 2 (3-Scheme 1) is overfunctionalized

R '= R' = H, then it can be functionalized by alkylation as needed. The sulfonamide is deprotonated using a base such as sodium hydride and then alkylated using an alkyl halide such as benzyl bromide or methyl iodide to form compound 2 of Scheme 2. The sulfonamide can then be alkylated a second time using sodium hydride and to form compound 3 of Scheme 2. It can then be converted to the desired urea using the method developed in Scheme 1.

Scheme 3

SO ₂ NR'R

O '

a) l) NCS, AcOH, H ₂ O II) NaOH, MeOH

b) H ₂ SO _4> HNO ₃ c) NaOH, MeOH d) PCI _5i POCI ₃

e) NHR'R, Et ₃ N

An alternative way for compound 5 - Scheme 3 (Compound 3 of Scheme 1) is given above in Scheme 3, where commercially available 2,6-dichlorothiol can be oxidized to sulfonyl halide using a halogenating agent such as NCS, NBS. chlorine or bromine in the presence of a protic solvent such as alcohol, acetic acid or water. The sulfonyl halide can be hydrolyzed using metal hydroxide such as sodium or potassium hydroxide to form the corresponding sulfonic acid salt. The sulfonic acid salt can then be nitrated under nitrating conditions as nitric acid in a solvent strong acid such as sulfuric acid to give nitrophenylsulfonic acid, compound 3 of scheme 3. This sulfonic acid can be converted to sulfonamide 5 - scheme 3 by uses a three - step process involving the formation of a metal salt using a base such as sodium hydroxide, sodium hydride or. sodium carbonate to form compound 4 of Scheme 3. The sulfonic acid salt is then converted to the corresponding sulfonamide using the desired amine HNR'R 'in triethylamine and at temperatures ranging from -78 ° C to 60 ° C to the corresponding sulfonamide 5 from Scheme 3 (Compound 3 from Scheme 1) was obtained. The sulfonamide 5 of Scheme 3 can be further processed by the methods given in Scheme 1. This method is not limited to 2,6-dichlorothiol, it can also be used for 2,6-dibromothiol and 2,6- diiodothiol. The halides of these compounds can be converted to the corresponding cyano, amino, thio, or alkoxy compounds by nucleophilic substitution reactions using nucleophiles such as alkylthiolates, alkoxides, amines and cyanides. The halides can be further functionalized by palladium coupling and carbonylation reactions well known in the art to form the corresponding amide, carbonyl, alkenyl, alkyl, phenyl and heterocyclyl substituted products as required by formula (I).

Examples of carrying out the invention

The invention will be described by the following examples, which are by way of illustration only, without limiting the scope of the invention. All temperatures are in degrees Celsius, all solvents are of the highest purity available, and all reactions are conducted under anhydrous conditions in argon, unless stated otherwise.

In the examples, the mass spectra were performed on a VG Zab mass spectrometer using fast atomic bombardment, unless otherwise stated. ^{The 1} H NMR (hereinafter, NMR ') spectrum was read at 250 MHz using a Bruker AM 250 or At 400 spectrometer.

Multiplicities mean: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, and br means broad signal. Sat. means a saturated solution, eq means an amount of one molar equivalent or reagent relative to the parent reagent.

General synthesis of 2-hydroxy-3-amino-6-chlorobenzenesulfonamide

a) 2,6-dichlorobenzenesulfonyl chloride

To a mixture of 200 ml of acetic acid, water and dibromomethane (3/1/4, v / v / v), 2,6-dichlorobenzentiol (10.0 g, 55.8 mmol) were added N-chlorosuccinimide (37.28 g, 279 mmol) and potassium acetate (2.29 g, 27.9 mmol). The resulting mixture was stirred at 0 ° C and then warmed to room temperature overnight. The mixture was then diluted with 200 ml of dichloromethane and washed with water (100 ml x 3). The organic layer was dried over sodium sulfate and concentrated to give the desired product (11g, 80%).

¹ NMR (CDCl ₃ ) δ 7.57 (d, 2H), 7.47 (t, 1H).

b) 2,6-dichlorobenzenesulfonamide

A solution of 2,6-dichlorobenzenesulfonyl chloride (10.50 g, 42.77 mmol) in 100 ml of pyridine was added dropwise to 100 ml of pyridine while passing anhydrous ammonia gas through the solution for 4 hours at 0 ° C. The mixture was acidified to pH> 1 with 6 N

to aqueous HCl, then extracted with ethyl acetate. The combined organic layer was dried over sodium sulfate and concentrated to give the desired product (8.69 g, 90%) EI-MS (m / z) 225.0, 227.1 (M ').

c) 2,6-dichloro-3-nitrobenzenesulfonamide

To a solution of 2,6-dichlorobenzenesulfonamide (7.8 g, 34.5 mmol) in 30 ml of concentrated sulfuric acid at 0 ° C was added dropwise nitric acid (1.74 ml, 41.4 mmol). The mixture was stirred at 0 ° C for 2 hours, then 200 ml of water was added to give a precipitate. The mixture was filtered. The white precipitate was separated, washed with water and dried under 0 vacuum to give the desired product (7.17 g, 76%).

NMR (DMSO-dj): δ 8.25 (s, 2H), 8.20 (d, 1 Η), 7.92 (d, 1Η).

d) 2-acetoxy-3-nitro-6-chlorobenzenesulfonamide

A solution of 2,6-dichloro-3-nitrobenzenesulfonamide (2.04 g, 7.5 mmol), potassium acetate (2.21 g, 22.5 mmol) and 18-crown-6 (5.95 g, 22.5 mmol) in 50 ml of dimethyl sulfoxide was heated at 45 ° In 7 days. The mixture was acidified with 1 N aqueous HCl and extracted with ethyl acetate. The organic layer was concentrated to give the crude product. Silica gel column chromatography eluting with ethyl acetate / hexane / acetic acid (50/49/1, v / v / v) afforded the desired product (1.67 g, 76%). EI-MS (m / z) • 293.1,295.1 (M ').

e) 2-hydroxy-3-nitro-6-chlorobenzenesulfonamide

A solution of 2-acetoxy-3-nitro-6-chlorobenzenesulfonamide (1.72 g, 5.83 mmol) chlorotrimethylsilane (2 ml) and fuming sulfuric acid (0.5 ml) in methanol was refluxed for 20 hours. The solvent was removed. The residue was diluted with ethyl acetate and washed with water. The organic layer was dried over sodium sulfate and concentrated to give the desired product (1.0 g, 68%) EI-MS (m / z) 251.1,253.2 (Μ ').

f) 2-hydroxy-3-amino-6-chlorobenzenesulfonamide

To a solution of 2-hydroxy-3-nitro-6-chlorobenzenesulfonamide (1.1 g, 4.36 mmol) in ethyl acetate was added 10% Pd / C (500 mt). Through the mixture

hydrogen was purged and then stirred under a hydrogen atmosphere for 4 hours at room temperature. The mixture was filtered through celite and the celite was washed with methanol. The solvent was evaporated to give the desired product (91%). EI-MS (m / z) 221.1, 223.1 (M ').

Example 1

A standard method for condensing aniline with isocyanate.

Preparation of N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 H -cyclohexyl urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (150 mg, 0.67 mmol) and cyclohexyl isocyanate (95 μl, 0.74 mmol) in 1 mL of dimethylformamide was stirred at room temperature for 20 hours. The mixture was diluted with ethyl acetate and washed with water to give the crude product. Purification by silica gel column chromatography, eluting with ethyl acetate / hexane (50/50, v / v) followed by recrystallization from acetone and hexane to give the desired product (121 mt, 52%) LC-MS (m / e) z) 348.0 (M ⁺ ).

Example 2

Preparation of N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 H (1-adamantyl) urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (120 mg, 0.54 mmol) and 1-adamantyl isocyanate (115 mg, 0.64 mmol) in 1 mL of dimethylformamide was stirred at room temperature for 20 hours. The mixture was diluted with ethyl acetate and washed with water to give the crude product. Purify by silica gel column chromatography eluting with ethyl acetate / hexane (50/50, v / v) to give the desired product (122 mg, 56%). LC-MS (m / z) 400.0 (M ⁺ ).

Example 3

Preparation of N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 H (tetrahydro-2-pyranyl) urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (170 mg, 0.76 mmol) and tetrahydro-2-pyranyl isocyanate (117 mg, 0.92 mmol) in 1.5 ml of N, N-dimethylformamide was stirred at room temperature for 20 hours. The mixture was diluted with ethyl acetate and washed with water to give the crude product. Purification by silica gel column chromatography eluting with ethyl acetate / hexane (60/40, v / v) followed by separation with Gilson HPLC gave the desired product (28 mg, 10%). LC-MS (m / z)

350.2 (M ⁺ ).

Example 4

A standard method for the synthesis of ureas by coupling carboxylic acids with aniline. Synthesis of N- [4-χπορο-2hydroxy-3-sulfamylphenyl] -N- (3-tetrahydrofuranyl) urea

To a solution of tetrahydrofuran-3-carboxylic acid (0.093 ml, 1.0 mmol) in dimethylformamide (0.5 ml) was added DPPA (0.25 ml, 1.2 mmol) and TEA (0.25 ml, 1.8 mmol) and the reaction mixture was heated at 70 After 18 hours, 3-amino-6-chloro-2-hydroxybenzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated to 70 °

C. After 18 hours water was added to the mixture and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 18 mg (5%) of N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N- (3-tetrahydrofuranyl) urea. LC-MS (m / z) 336 (M ⁺ ).

Example 5

Synthesis of 6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) -ureido] benzenesulfonamide

To a solution of 2-methyl-cyclopropanecarboxylic acid (0.097 ml, 1.0 mmol) in dimethylformamide (0.5 ml) was added DPPA (0.25 ml, 1.2 mmol) and TEA (0.25 ml, 1.8 mmol) and the reaction mixture was heated at

90 ° C. After 18 hours, 3-amino-6-chloro-2-hydroxybenzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated to 90 °

C. After 18 hours water was added to the mixture and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 29 mg (11%) of 6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) ureido] -benzenesulfonamide. LC-MS (m / z) 320 (M ⁺ ).

Example 6

Synthesis of 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethylcyclopropyl) -ureido] -benzenesulfonamide

To a solution of 2,2,3,3-tetramethyl-cyclopropanecarboxylic acid (0.142 mL, 1.0 mmol) in dimethylformamide (0.5 mL) was added DPPA (0.25 mL, 1.2 mmol) and TEA (0.25 mL, 1.8 mmol) and the reaction mixture. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated to 80 ° C. After 18 hours water was added to the mixture and extracted with ethyl acetate. . The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 96 mg (11%) of 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethylcyclopropyl) -ureido] -benzenesulfonamide. LC-MS (m / z) 347 (M ⁺ ).

Example 7

Synthesis of 6-chloro-2-hydroxy-3- (3-piperidin-4-yl-ureido) benzenesulfonamide

To a solution of piperidine-1,4-dicarboxylic acid mono-tert -bugyl ester (0.23 mL, 1.0 mmol) in dimethylformamide (0.5 mL) was added DPPA (0.215 mL, 1.0 mmol) and TEA (0.139 mL, 1.0 mmol) and The reaction mixture was heated at 80 ° C. After 18 hours, 3amino-6-chloro-2-hydroxy-benzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated at 80 ° C. After 18 hours water was added and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 12 mg of 6-chloro-2-hydroxy-3 (3-piperidin-4-yl-ureido) -benzenesulfonamide. LC-MS (m / z) 349 (M ⁺ ).

Example 8

Synthesis of N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '- (4-methylcyclohexyl) urea

To a solution of 4-methyl-cyclohexanecarboxylic acid (0.141 ml) in dimethylformamide (0.5 ml) was added DPPA (0.215 ml) and TEA (0.139 ml) and the reaction mixture was heated at 80 ° C. After 18 hours, 3-amino was added. -6-chloro-2-hydroxy-benzenesulfonamide (0.150 g) and the reaction mixture was heated at 80 ° C. After 18 hours water was added to the mixture and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 15 mg of 1 N- [4-chloro-2-hydroxy-3 sulfamylphenyl] -N '- (4-methyl-cyclohexyl) urea. LC-MS (m / z) 347 (M ⁺ ).

Example 9

Synthesis of 6-chloro-2-hydroxy-3- [3- (3-methoxy-cyclohexyl) ureido] -benzenesulfonamide

To a solution of 3-methoxy-cyclohexanecarboxylic acid (0.143 ml) in dimethylformamide (0.5 ml) was added DPPA (0.25 ml) and TEA (0.25 ml) and the reaction mixture was heated at 70 ° C. After 18 hours, 3-amino was added. -6-chloro-2-hydroxy-benzenesulfonamide (0.222 g) and the reaction mixture was heated at 70 ° C. After 18 hours water was added to the mixture and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The crude residue was purified by HPLC to give 3 mg of 6-chloro-2-hydroxy-3- [3 (3-methoxy-cyclohexyl) -ureido] -benzenesulfonamide. LC-MS (m / z) 378 (M ⁺ ).

Example 10

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1'-cyclopentyl urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (200 mg, ** 0.90 mmol) and cyclopentyl isocyanate (100 mg, 0.90 mmol) in 1.5 ml Ν dimethylformamide was stirred at room temperature for 20 hours. Purification by silica gel column chromatography eluting with ethyl acetate / hexane (20/80 v / v) and subsequent recrystallization from diethyl ether and hexane gave the desired product (130 mg, 43%). LCMS (m / z) 334.2 (M ⁺ ).

Example 11

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1'-cyclobutyl

urea

Under argon, a mixture of cyclobutanecarboxylic acid (250 mg, 2.50 mmol), diphenylphosphoryl azide (756 mg, 2.74 mmol) and triethylamine (0.38 ml, 2.74 mmol) in 3 ml N, N-dimethylformamide was heated at 80 ° C for 2 hours. The mixture was cooled to room temperature and 3-amino-6-chloro-2-hydroxybenzenesulfonamide (556 mg, 2.50 mmol) was added. The resulting mixture was stirred at room temperature for 20 hours. Purification by Gilson HPLC eluting with acetonitrile / water (10/90, v / v for 10 minutes) gave the desired product (137 mg, 17%). LC-MS (m / z) 320.0 (M ⁺ ).

Example 12

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclopropanyl urea

Under argon, a mixture of cyclopropanecarboxylic acid (500 mg, 5.81 mmol), diphenylphosphoryl azide (1.58 g, 5.81 mmol) and triethylamine (0.81 mL, 5.81 mmol) in 3 mL N, N-dimethylformamide was heated at 80 ° C for 3 hours. The mixture was cooled to room temperature and 3-amino-6-chloro-2-hydroxybenzenesulfonamide (200 mg, 0.89 mmol) was added. The resulting mixture was stirred at room temperature for 20 hours. Purification by Gilson HPLC eluting with acetonitrile / water (10/90, v / v to 90/10 v / v for 10 minutes) afforded the desired product (74 mg, 27%). LC-MS (m / z) 306.2 (M ⁺ ).

Example 13

N- (3- (M'-Boc-piperazine) -aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclopentyl urea

A solution of 3-N- (Boc-piperazine) amino-6-chloro-2-hydroxy benzenesulfonamide (200 mg, 0.51 mmol) and cyclopentyl isocyanate (57 μl, 0.51 mmol) in 2 ml of N, N-dimethylformamide was stirred at room temperature. temperature for 20 hours. Purification by silica gel column chromatography eluting with ethyl acetate / hexane (30 / 70p v / v) followed by recrystallization from dichloromethane and hexane gave the desired product (180 mg, 70%). LC-MS (m / z) 503.2 (M ⁺ ).

Example 14

1 \ 1- (3-M'-piperazine) -aminosulfonyl-4-chloro-2-hydroxyphenyl) -I-cyclopentyl urea hydrochloride

A solution of N- (3- (1 N-Boc-piperazine) aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclopentyl urea (140 mt, 0.27 mmol) in 2 ml of 4N HCl in 1,4-dioxane was stirred at room temperature for 1 hour. The mixture was concentrated. Recrystallization from acetonitrile gave the desired product (105 mg, 88%). LC-MS (m / z) 403.2 (M ⁺ ).

Example 15

N- (3- (M'-Boc-piperazine) -aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclobugyl urea

Under argon, a mixture of cyclobuganecarboxylic acid (500 mg, 4.99 mmol), diphenylphosphoryl azide (1.18 g, 5.49 mmol) and triethylamine (0.76 ml, 5.49 mmol) in 3 ml N, N-dimethylformamide was heated at 80 ° C for 2 hours. The mixture was cooled to room temperature and 3-N- (Bocpiperazine) amino-6-chloro-2-hydroxybenzenesulfonamide (1.95 g, 4.97 mmol) was added. The resulting mixture was stirred at room temperature for 20 hours. Purification by silica gel column chromatography eluting with ethyl acetate / hexane (30/70, v / v to 50/50 v / v) afforded the desired product (500 mg, 21%) LC-MS (m / z) 489.2 (M ⁺ ).

Example 16

Synthesis of N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N '- [(1c,

28) - (-) - 2-benzyloxycyclohexyl] urea (1c, 28) - (-) 2-benzyloxycyclohexyl isocyanate

Under argon, a solution of (18,28) - (-) 2-benzyloxycyclohexylamine (100 mg, 0.49 mmol) in 5 ml of dichloromethane was added to a solution of di-tert-butyl dicarbonate (148 mg, 0.68 mmol) and 3-dimethylaminopyridine ( 87 mg, 0.49 mmol) in 5 ml of dichloromethane. The mixture was stirred for 15 min at room temperature, then concentrated to give the crude product. FTIR: 2237.92 cm ' ¹ .

N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 N - [(18,28) - (-) 2-benzyloxycyclohexyl] urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (109 mg, 0.49 mmol) and crude (18,28) - (-) 2-benzyloxycyclohexyl isocyanate in 2 ml of N, N-dimethylformamide was stirred at room temperature for 20 hours . Purification by Gilson HPLC eluting with acetonitrile / water (10/90, v / v to 90/10 v / v for 10 minutes) afforded the desired product (60 mg, 27% of the two steps). LC-MS (m / z) 454.0 (M ⁺ ).

Example 17

Synthesis of N- (3- (N'-dimethyl) - (aminosulfonyl-4-chloro-2-hydroxyphenyl) - N'-cyclobutyl urea hydrochloride (3- (N-dimethyl) -aminosulfonyl-4-chloro-2-hydroxyphenyl) -No Boc-H “-cyclobutyl urea

Under argon, a mixture of cyclobutylamine (14 μl, 0.16 mmol), trimethylaluminum (80 μl, 0.16 mmol) in 3 mL dichloromethane was stirred at room temperature for 10 minutes. A solution of 7- (N-dimethyl) aminosulfonyl-3-Boc-5-chloro-2-benzoxazolinone (60 mg, 0.16 mmol) in 3 ml of dichloromethane was added. The resulting mixture was stirred at room temperature for 20 hours. Purification by silica gel column chromatography eluting with ethyl acetate / hexane (20/80 v / v) afforded the desired product (36 mg, 51%). LC-MS (m / z) 448.2 (M ⁺ ).

N- (3- (N'-dimethyl) - (aminosulfonyl-4-chloro-2hydroxyphenyl) - LH-cyclobugyl urea hydrochloride

To a solution of N- (3- (1'-dimethyl) -aminosulfonyl-4-chloro-2 'hydroxyphenyl) -H-Boc-N' -cyclobutyl urea (36 mg, 0.08 mmol) in 2 ml of 4N HCl in 1, 4-dioxane was stirred at room temperature for 1 hour. The mixture was concentrated. Recrystallization from acetone and hexane gave the desired product (20 mg, 65%). Elemental analysis in% gives:

calculated: C 44.89; H, 5.22; N, 12.08;

found: C 44.85; H, 5.00; N11.91.

Use

The compounds of formula (I) or a pharmaceutically acceptable salt thereof may be used in the manufacture of a medicament for the prevention and treatment of a disease in a human or other mammal that is exacerbated or induced by excessive or unregulated IL-8 cytokine production by such mammals cells, including, but not limited to, monocytes and / or macrophages, or

other chemokines that bind to the IL-8α or β receptor, also referred to as type I or type-1 receptor.

Accordingly, the invention provides the use of the compounds of the invention for the preparation of a pharmaceutical composition for the treatment of a chemokine mediated disease, wherein the chemokine is such that it binds to the IL-8α or β receptor, wherein an effective amount of a compound of formula ( I) or a pharmaceutically acceptable salt thereof. In particular, the chemokines are IL-8, GROa, GROp, GROy, NAP-2 and ENA-78.

The compounds of formula (I) are administered in an amount sufficient to inhibit cytokinin function, in particular IL-8,

GROa, GROp, GROy, NAP-2 and ENA-78, so that they are biologically regulated down to normal levels and thus improve the disease state. Abnormal levels of IL-8, GROa, GROp, GROy, NAP-2 and ΕΝΑ-

for example, in the context of the present invention, represent: (i) levels of free IL-8 greater than or equal to 1 picogram per ml; (II) any cells associated with IL-8, GROa, GROp, GROy, NAP-2 and ENA-78 above normal physiological levels, or (III) the presence of IL-8, GROa, GROp, GROy, NAP-2 and ENA- 78 above baseline levels in cells or tissues in which IL-8, GROa, GROp, GROy, NAP-2 and ENA-78 are respectively produced.

The compounds of formula (I) have generally been shown to have an elongated t _1/2 and improved oral bioavailability compared to the compounds described in WO 96/25157 and WO 97/29743, the contents of which are incorporated herein by reference.

There are many conditions in which excessive or unregulated production of IL-8 is involved in the worsening or causing of the disease. Chemokine mediated diseases include psoriasis, atopic dermatitis, osteoarthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative, ulcerative, ulcerative, , gram-negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft rejection, Alzheimer's disease, allograft rejection, malaria, restenosis, angiogen anger, atherosclerosis, osteoporosis, gingivitis, and unwanted hematopoietic stem cell release and illness caused by respiratory viruses, herpes viruses and hepatitis, tingling, tingling, tingling, tingling arthritis, herpes, encephalitis, vasculitis of the central nervous system, traumatic brain damage, tumors of the central nervous system, subarachnoid hemorrhage, trauma after surgery, interstitial Mon. eutonitis, hypersensitivity, crystalline arthritis, acute and chronic pancreatitis, chronic alcoholic hepatitis, necrotizing enterocolitis, chronic sinusitis, acne, gastric ulcers, acupuncture airways obliterating bronchitis organizing pneumonia, bronchiectasis, obliterating bronchiolitis, chronic bronchitis, cor pulmonae, dyspnoea, emphysema, hypercapnia, hyperinflation, hypoxemia, hyperoxia-induced scarlet, hypoxia, surgical reduction of the volume of the lung, pulmonary fibrosis, pulmonary hypertension, right ventricular hypertrophy, sarcoidosis, Crohn's small airways fan perfusion mismatching, wheeze, colds and lupus.

These diseases are initially characterized by massive neutrophil infiltration, T-cell infiltration or neovascular growth and are associated with increased production of IL-8, GROa, GROp, GROy, NAP-2 or ENA-78, which is responsible for the chemotaxis of neutrophils of the site of inflammation or the targeted growth of endothelial moles. Unlike other inflammatory cytokines (IL-1, TNF and IL-6), IL-8, GROa, GROp, GROy, NAP-2 or ENA-78 have the unique ability to promote non-neutrophil chemotaxis, an enzymatic release including elastase release, without be limited to it, as well as superoxide production and activation. α-Chemokines, and in particular GROa, GROp, GROy, NAP-2, or ENA-78, working through IL-8 type I or II receptors, can promote neovascularization of tumors through targeted growth of endothelial cortices. Therefore, inhibition of IL-8 induced chemotaxis or activation will result in a decrease in neutrophil infiltration.

Evidence lately suggests the role of chemokines in the treatment of HIV infection, Littleman et al., Nature 381, pp. 661 (1996) and Koup et al., Nature 381, pp.667 (1996).

Recently, there is also evidence for the treatment of atherosclerosis with the use of IL-8 inhibitors. The first communication, Boisvert et al., J. Clin. Invest, 1998, 101: 353-363 shows by bone marrow transplantation that the absence of IL-8 receptors on stem cells (and therefore on monocytes / macrophages) leads to a decrease in the development of atherosclerotic plaques in LDL receptor deficient mice. Further sources supporting this claim are: Apostolopoulos et al., Arterioscler. Thromb. Yours, Biol. 1996, 16: 1007 1012; Liu, et al., Arterioscler. Thromb. Yours, Biol. 1997.17: 317 - 323; Rus, et al., Atherosclerosis, 1996, 127: 263-271; Wang et al., J. Biol. Chem., 1996, 271: 8837-8842; Yue, et al., Eur. J. Pharmacol., 1993, 240: 81-84; Koch et al., Am. J. Pathol., 1993,142: 1423 - 1431; Lee et al., Immunol. Lett. 1996, 53, 109-113; Terkeltaub et al., Arterioscler. Thromb., 1994, 14: 47 - 53.

The invention also provides receptor antagonist compounds of formula (I) as treatment agents, in the acute phase, and for the protection of those individuals who are considered to be susceptible to CNS lesions.

CNS injuries as defined herein include open and penetrating head injuries such as surgery or closed head trauma, as with head area injuries. This definition also includes ischemic stroke, especially in the brain.

Ischemic stroke can be defined as focal neurological disorder resulting from insufficient blood supply to a specific brain region, usually as a result of embolism, thrombus, or local atheromatic closure of a blood vessel. The role of inflammatory cytokines in this field and the present has been revealed

the invention provides a means of effectively treating these disorders. Relatively few treatments have so far been available for acute damage like this.

TNF-α is a cytokine with pro-inflammatory effects, including endothelial leukocyte adhesion molecular expression. Leukocytes infiltrate ischemic brain lesions, and hence compounds that inhibit or reduce TNF levels will be useful in treating ischemic brain damage. See Lui et al., Stroke, Vol. 25., No. 7, pp. 1481-88 (1994), the description of which is incorporated herein by reference.

Models of closed head injury and treatment with mixed 5-LO / CO agents have been discussed by Shohami et al., J of Vaisc & Clinical Physiology and Pharmacology, Vol.3, No. 2, pp. 99 - 107 (1992), incorporated herein by reference. Treatment resulting in a decrease in edema formation has been found to improve functionally the condition of the animals treated.

The compounds of formula (I) are administered in sufficient amount to inhibit IL-8 binding to IL-8 alpha or beta receptors, as evidenced by a decrease in neutrophil chemotaxis and activation. The finding that compounds of formula (I) are inhibitors of IL-8 binding is based on the effects of the compounds of formula (I) in the in vitro receptor binding assays described herein. The compounds of formula (I) have been shown to be type II inhibitors of IL-8 receptors.

As used herein, the term "mediator of IL-8 diseases or disease states" refers to some or all of the disease states in which IL-8, GROa, GROp, GROy, NAP-2 or ENA-78 play a role either by producing IL- 8, GROa, GROp, GROy, NAP-2 or ENA-78 or as IL-8, GROa, GROp, GROy, NAP-2 or ENA-78 induce the release of another monokine such as IL-1, IL-6 or TNF without being limited to them. A disease state in which, for example, IL-1 is a major component and whose production or action is increased or increased

j secreted as a response to IL-8 would be considered a disease condition mediated by IL-8.

As used herein, the term, β mediator of IL-8 diseases or disease states refers to all disease states in which a chemokine that binds to IL-8 alpha or beta receptors plays a role, either through the production of IL-8 itself. or IL-8 induces, but is not limited to, the release of another monokine such as IL-1, IL-6 or TNF. A disease state in which, for example, IL-1 is a major component and whose production or action is increased or secreted in response to IL-8 would be considered a disease condition mediated by IL-8.

As used herein, the term ditokine "refers to any secreted polypeptide that affects cell function and is a molecule that modulates interactions between cells in an immune, inflammatory or hematopoietic response. The cytokine includes, but is not limited to, monokines and lymphokines, regardless of what cells produce them. For example, monokine is thought to be generally obtained as the production and secretion of a mononuclear cell such as a macrophage and / or monocyte. However, many other cells produce monokines, such as natural killer cells, fibroblasts, basophils, neugrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epidermal keratinocytes and B-lymphocytes. Lymphokines are generally 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-alpha (TNF-α), and tumor necrosis factor-beta (TNF-β).

The term "chemokine" used herein refers to any secreted polypeptide that affects cell function and is a molecule that modulates interactions between cells in an immune, inflammatory or hematopoietic response, similar to the term "cytokine" above.

Chemokine is primarily secreted through vows transmembranes and causes chemotaxis and activation of the specific ^^ n ^ Biały blood cells and leukocytes, neutrophils, monocytes, macrophage _h l t cells, B-cells, endothelial cells and smooth muscle cells. * 'riMOP ^{and chemokines} include, but are not limited to, IL-8, GR <\ _4> ο | 1Οβ, NAP-2, ENA-78, IP-10, ΜΙΡ-1α, ΜΙΡ-β, PF4 and MCP 1,2 and 3.

In order to use a compound of fsomula (I) or a pharmaceutically acceptable salt thereof in treatment, it is normal for it to be formulated in a pharmaceutical composition in accordance with standard pharmaceutical practice. The invention also relates to a pharmaceutical composition comprising 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, may be administered by any of the commonly used routes such as, for example, orally, externally, parenterally or by inhalation. The compounds of formula (I) can be administered in conventional dosage forms by combining a compound of formula (I) with standard phoamacetamide carriers, as is customary. Compounds of oiomula (I) may also be administered in conventional doses in combination with a h-tsel, second active compound. Common practice involves grinding. granulating and compressing or dissolving the ingredients, according to the need to obtain the desired formulation. It is understood that the form and nature of the pharmaceutical carrier or diluent is determined by the amount of active ingredient with which it is to be combined, by the mode of administration and other well known factors. The carrier (s) must be compatible with the other ingredients of the formulation- ^? not harmful to his receiver.

The pharmaceutical carrier used, for example, must be either solid or liquid. Examples of solid carriers are lactose, white soil, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Examples of liquid carriers are syrup, peanut butter, olive oil, water and more. The carrier or diluent may also include a retardant well known in the art, such as glyceryl monostearate or glyceryl distearate, alone or together with wax.

A wide variety of pharmaceutical forms can be used. Thus, if a solid carrier is used, the formulation may be tableted, placed in a hard gelatin capsule in powdered or pelleted form or in the form of pastilles or lozenges. The amount of solid carrier may vary greatly, but is preferably from about 25 mg to about 1 g. When using a liquid carrier, the preparation will be in the form of a syrup, emulsion, soft gelatin capsules, sterile injection fluid in, for example, an ampoule or a non-aqueous liquid suspension.

The compounds of formula (I) may be administered externally, i. unsystematically. This involves administering a compound of formula (I) externally to the epidermis or oral cavity, or installing a compound in the ear, eye or nose so that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal, and intramuscular administration.

Formulations suitable for external use include liquid or semi-liquid preparations suitable for penetration into the skin at the site of inflammation such as liniments, lotions, creams or pastes and drops suitable for use in the eyes, ears or nose. The active ingredient may, when applied externally, be from 0.001 to 10% w / w, for example from 1% to 2% by weight of the formulation. However, it may be present up to 10% w / w, preferably less than 5%, better than 0.1 to 1% w / w in the formulation.

According to the present invention, lotions are a formulation for application to the skin or eyes. An eye lotion may include a sterile aqueous solution, optionally containing a bactericidal agent, and may be prepared by methods similar to those used in the preparation of drops. Lotions and liniments for application to the skin may also contain a means of accelerating drying on the skin and of cooling the skin such as alcohol or acetone and / or a moisturizer such as glycerol or any oil such as castor or peanut.

Creams, ointments or pastes according to the invention are semi-solid formulations of the active ingredient for external use. They can be made by mixing the active ingredient in finely divided or powdered form, alone or in solution or suspension in aqueous or non-aqueous liquid, using an appropriate technique, oil or non-oil based. The base may contain hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax or metal soap; lanolin or its derivatives or fatty acids such as stearic or oleic acid together with an alcohol such as propylene glycol or macrogel. the formulation may contain any suitable surfactant such as anionic, cationic or non-ionic, for example sorbitan ester or polyoxyethylene derivatives thereof. Suspension agents such as natural resins, cellulose derivatives or inorganic substances such as silicon derivatives and other ingredients such as lanolin may also be included.

The droplets according to the invention may include sterile aqueous or oily solutions or suspensions and may be obtained by dissolving the active ingredient in a suitable aqueous solution of

 a bactericide and / or fungicide 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 maintained at 98-100 ° C for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to a 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 oily solutions are glycerol, dilute alcohol and propylene glycol.

The compounds of formula (I) may be administered parenterally, i. by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. 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) can be administered by inhibition, i.e. by intranasal and oral inhalation. Suitable dosage forms for such administration as aerosol formulations or metered doses for inhalation may be prepared by conventional techniques.

For all applications of the compounds of formula (I), the daily oral dosage regimen will preferably be from about 0.01 to about 80 mg / kg total body weight. The daily parenteral dose is about 0.01 to about 80 mg / kg of total body weight. The daily dosage regimen for external administration is from about 0.1 mg / kg to about 150 mg, administration one to four, preferably two to three times daily. The daily inhalation dose will preferably be from about 0.01 mg / kg to about 1 mg / kg per day. It will also be appreciated that one of ordinary skill in the art will &apos; determine the optimum amount and intervals of administration of the individual doses of the compound of formula (I) or a pharmaceutically acceptable salt thereof, having regard to the nature and severity of the condition being treated, the site of administration as well as the particular patient being treated and this optimum can be determined using conventional techniques. Also, one skilled in the art will be able to determine the optimal course of treatment, i. the number of doses of the compound of formula (I) or a pharmaceutically acceptable salt thereof per day, the number of days, etc.

The invention will be described by the following biological examples, which are merely illustrative and are not intended to be organized in any way.

Biological examples

The IL-8 and GROa chemokine inhibitory effects of the compounds of the invention were determined by the following in vitro assay:

Receptor Binding Test:

[ ¹²⁵ l] IL-8 (human recombinant) was prepared from Amersham Corp., Arlington Heights, IL, with a specific activity of 2000 Ci / mmol. GROa is obtained from NEN-New England Nuclear. All other chemicals are of analytical purity. High levels of recombinant human IL-8 type alpha or beta receptors are individually expressed in Chinese hamster ovary cells, as previously described (Holmes, et al., Science, 1991, 253,1278). The Chinese hamster ovary membrane was homogenized according to the procedure described previously (Haour, et al., J. Biol. Chem., 249, pp. 2195 - 2205 (1974)). Homogenisation buffer alone was changed to 10 mM Tris-HCl, 1 mM MgSO ₄ , 0.5 mM EDTA (ethylenediaminetetraacetic acid), 1 mM PMSF (α-toluenesulfonyl fluoride), 0.5 mg / l levopeptin, pH 7.5. The membrane protein concentration was determined using Pierce Co. micro-test device using bovine serum albumin as standard.

ΐ

All tests were performed in a 96 well microplate. Each reaction medium contained ¹²⁵ l of IL-8 (0.25 nM) or ¹²⁵ l of GROa and 0.5 µt / ml of IL-8Ra or 1.0 µg / ml of IL-8Rp membranes in 20 mM bis-trispropane and Ok | three HCl buffers, pH 8.0, containing 1.2 mM MgSO ₄ , 0.1 mM EDTA, γ mM NaCl and 0.03% CHAPS. In addition, added represents ^ z _0T0 interest drug or a compound which has previously solution ^ _{14 O in} DMSO so as to contain a final concentration of between 0.01 nM iqq uM. The test is initiated by the addition of ¹²⁵ l of IL-8. After 1 _EXAMPLE room temperature the plate was harvested using Tomte ^ gg. well harvester on a glass fiber filter layer blocked with ¢ - ·% polyethyleneimine / 0.5% BSA and washed three times with 25 mM NaCl, 1 µM m TrisCl, 1 mM MgSO ₄ , 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. After γο _{Β3, the} filter was washed and counted on a Betaplate liquid scintillation day counter. Recombinant IL-8Rα or type I receptor is referred to herein as an illicit receptor, and recombinant IL-8Rβ or type II receptor is referred to herein as an authorized receptor.

Representative compounds of formula (I), Examples 1 to Ί06, showed positive inhibitory activity in this assay at 1 C ₅₀ levels <30 μM.

Chemotactic tests:

The in vitro inhibitory properties of these compounds were determined in the neutrophil chemotaxis assay as described in Current Protocols in Immunology, Vol. I, Suppl. 1, Unit 6.12.3., The entirety of which is incorporated herein by reference. Neutrophils are isolated from human blood as described in Current Protocols in Immunology, Vco I. Suppl. 1, Unit 7.23.1, the description of which in its entirety is incorporated by reference. The chemoattractants IL-8, GROa, GROp, GROy, and NAP-2 were placed in the bottom chamber of a 48 multi-well chamber (Ν ^^ Probe, Cabin John, MD) at concentrations between 0.1 and 100 nM. The two stones * £ 2 * are separated by a 5 μm polycarbonate filter. When drunk + ^ ^t ύ ·

compounds of the invention, they were mixed with the cells (0.001-1000 nM) just before the addition of cells to the upper chamber. The incubation was allowed to proceed between about 45 and 90 minutes at about 37 ° C in a humidified incubator with 5% CO ₂ . At the end of the incubation period, the polycarbonate membrane was removed and the top was washed, then the membrane was stained using a Diff Quick staining method (Baxter Products, McGaw Park. IL, USA). The chemotaxis cells to the chemokine were counted visually using a microscope. Generally, four fields are counted for each sample; these numbers are averaged to obtain the average number of cells that have migrated. Each sample is tested in three replicates and each compound is repeated at least four times. Some cells (positive control cells) are not added to the compound; these cells represent the maximum chemotactic response of the cells. When a negative control (unstimulated) is desired, no chemokine is added to the bottom chamber. The difference between the positive control and the negative control is the chemotactic activity of the cells.

Elastase release test:

The compounds of the invention are tested for their ability to protect the elastase release from human neugrophils. Neutrophils are isolated from human blood as described in Current Protocols in Immunology, Vol. I, Suppl. 1, Unit 7.23.1. PMNs 0.88 x 10 ⁶ cells suspended in Ringer's solution (Na 118, KCI 4.56, NaHCO ₃ 25, KH ₂ PO ₄ 1.03, glucose 11.1, HEPES 5 mM, pH 7.4) were placed in each well in a 96 well plate in volume of 50 μl. To this plate was added the test compound (0.001 -1000 nM) in a volume of 50 µl, Cytochalasin B in a volume of 50 µl (20 µg / ml) and Ringer buffer in a volume of 50 µl. These cells were allowed to warm (37 ° C, 5% CO ₂ , 95% RH) for 5 minutes before IL-8, GROa, GROp, GROy or NAP-2 was finally added.

concentration of 0.01 - 1000 nm. The reaction was allowed to proceed 45 minutes before 96 well plates were centrifuged (800 x g for 5 minutes) and 100 μl of supernatant removed. This supernatant was added to a second 96 well plate followed by an artificial elastase substrate (MeOSuc-Ala-Ala-Pro-Val-AMC, Nova Biochem, La Jolla, CA) to a final concentration of 6 μg / ml dissolved in phosphate buffered saline. Immediately place the plate in a fluorescent 96 well plate reader (Cytofluor 2350, Millipore, Bedford, MA) and read the data at 3 minute intervals according to the method of Nakajiama et al., J. Biol. Chem., 254, 4027 (1979). The amount of elastase released by PM Ns was calculated by measuring the degradation rate of MeOSuc-Ala-Ala-Pro-Val-AMC.

TNF-α in a brain injury trial

The present test verifies the expression of tumor necrosis factor mRNA in specific brain regions after experimentally induced lateral cerebrospinal injury resulting from traumatic concussion in rats (TBI). Adult Sprague - Dawley rats (n = 42) were anesthetized with sodium phenobarbital (60 mg / kg, ip) and subjected to lateral fluid cerebral damage following a moderate severity concussion (2.4 atm) centered above the left temporaparietapen cortex (cortex) ) or "sham" effect (anesthesia and surgery without damage n = 18). Animals are killed by decapitation 1, 6, or 24 hours after injury, the brains are removed and tissue samples are drawn from the left (damaged) parietal cortex (LC), the corresponding area in the contralateral right cortex (RC), from the cortex adjacent to the damaged parietal cortex (LA), the corresponding adjacent area in the right cortex (RA), from the left hippocampus (LH) and from the right hippocampus (RH). Total RNA was isolated and Northern blot hybridization was performed and quantified with respect to TNF-α positive control RNA (macrophage

- 100%). There was a marked increase in TNF-α mRNA expression in LH (104 ± 17% of positive control, p <0.05 vs. sham), LC (105 ± 21%, p <0.05) and LA (69 ± 8%, p <0.01) in the injured hemisphere 1 hour after injury. Increased expression of TNF-α mRNA was also observed in LH (46 ± 8%, p <0.05), LC (30 ± 3%, p <0.01) and LA (32 ± 3%, p <0.01) on the 6th hour, which is transmitted to the 24th hour after the injury. In the contralateral hemisphere, TNF-α mRNA expression increased in RH (46 ± 2%, p <0.01), RC (4 ± 3%), and RA (22 ± 8%) at 1 h and RH (28 ± 11%), RC (7 ± 5%) and RA (26 ± 6%, p <0.05) at the 6th hour but not at the 24th hour after injury. In sham (untreated surgery) or untreated animals, no logical changes in TNF-α mRNA expression were observed in any of the 6 brain panels in any of the hemispheres at any time. These results indicate that, after a lateral fluid-brain injury resulting from a traumatic concussion, the temporary expression of TNF-α mRNA changes in specific brain regions, including those where there is no hemisphere injury. Since 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 an important role in both acute and regenerative response to central nervous system (CNS) injury.

CNS-damaging model for IL-β mRNA

This trial characterized the regional expression of interleukin-1 β (IL-Ιβ) mRNA in specific brain regions after experimental lateral cerebrospinal injury resulting from traumatic concussion in rats (TBI). Adult Sprague - Dawley

. * {rats (η = 42) were anesthetized with sodium phenobarbital (60 mg / kg, ip) and subjected to lateral fluid-brain damage resulting from moderate-severity concussion (2.4 atm) centered over the left temporal parietal cortex (n = 18). ) or .sham exposure (anesthesia and surgery without damage n = 18). Animals are killed 1, 6, and 24 hours after injury, the brains are removed and tissue samples are drawn from the left (damaged) parietal cortex (LC), the corresponding area in the contrauterine right cortex (RC), from the cortex adjacent to the damaged parietal cortex ( LA), the corresponding adjacent area in the right cortex (RA), from the left hippocampus (LH) and from the right hippocampus (RH). Total RNA was isolated and Northern blot hybridization was performed and the amount of IL-Ιβ mRNA present in brain tissue was determined as a percentage of the relative radioactivity of IL-Ιβ positive macrophage RNA loaded on the same gel. One hour after injury, there was a marked increase in the expression of IL-RNAβ mRNA in LC (20.0 ± 0.7% of positive control, n = 6, p <0.05 versus sham animal), LH (24.5 ± 0.9%, p <0.05). and LA (21.5+ 3.1%, p <0.05) in the injured hemisphere, which remain elevated up to 6 hours after damage in the LC (4.0 ± 0.4%, n = 6, p <0.05) and LH (5.0 + 1.3%, p < 0.05). In sham or untreated animals, no expression of IL-Ιβ mRNA was observed in any of the corresponding brain spaces. These results suggest that following TBI, the temporary expression of IL-Ιβ mRNA is regionally stimulated in specific brain regions.

These regional changes in cytokines such as IL-Ιβ play a role after trauma.

All publications, including, but not limited to, patents and patent disclosures cited in the description, are incorporated herein by reference.

Ί

- -r

The foregoing description fully discloses the invention, including preferred embodiments thereof. Modifications and improvements to what is described herein are within the scope of the following claims. Without further processing, it is believed that a person skilled in the art, using the d description, can make the most of it. Therefore, the examples included herein are for illustrative purposes only and are not intended to limit the scope of the invention in any way.

Claims (13)

Claims in which: R _b is independently selected from the group consisting of hydrogen, NR ₆ R ₇ , OH, OR _a , Ci. ₅ alkyl, aryl, arylC _1-4 alkyl, arylC ₂ . ₄ alkenyl; cycloalkyl, cycloalkyl C _1-5 alkyl, heteroaryl, heteroarylC 1-6 alkyl, heteroarylC ₂ . ₄ alkenyl, heterocyclyl, heterocyclyl _m alkyl, heterocyclylC _{2. 4} alkenyl, all of these groups may optionally be substituted one to three times independently of the group consisting of halogen, nitro, halogen-substituted C 1-6 alkyl, C _1-4 alkyl, amino, mono or di-C _1-4 alkyl substituted amine, OR _a , C (O ) R _a , NR _a C (O) OR _a , OC (O) NR ₆ R ₇ , hydroxy, NR ₉ C (O) R _a , S (O) _m .R _a , C (O) NR ₆ R ₇₁ C (O) OH, C (O) OR _a , S (O) ₂ NR ₆ R ₇ , NHS (O) ₂ R _a, or the two R _b substituents together form a 3-10 membered ring, optionally substituted and containing in addition to carbon independently from 1 to 3 groups selected from NR _a , O, S, SO and SO ₂ and wherein the substituent may be unsaturated; R _a represents an alkyl, aryl, arylC _b4 alkyl, heteroaryl, heteroarylC ^ alkyl, heterocyclyl, COOR ₁₃ and heterocyclylC _m alkyl, all these groups can be optionally substituted; m is a number from 1 to 3; ηϊ is 0 or a number from 1 to 2; η is a number from 1 to 3; q is 0 or a number from 1 to 10; , ί t is 0 or a number from 1 to 2; s is a number from 1 to 3; Rf independently represents hydrogen, halogen, nitro, cyano, R _10-10 alkyl, halogen substituted R _6-10 alkyl, R ₂ . ₁₀ alkenyl, R _V10 alkoxy, halogen substituted R _V10 alkoxy, azide, S (O) _t R ₄ , (CR ₈ R ₈ ) _q S (O) _t R ₄ , hydroxy, hydroxy substituted C _1-4 alkyl, aryl, aryl C _m alkyl, aryl R ₂ . ₁₀ alkenyl, aryloxy, arylC 1-4 alkyloxy, heteroaryl, heteroarylalkyl, heteroaryl R ₂ . ₁₀ alkenyl, heteroarilR _V4 alkyloxy, heterocyclyl, heterocyclyl-alkyl, heterocyclyl-alkyloxy, heterocyclyl _{2. 10} alkenyl, (CR ₈ R ₈ ) _q NR ₄ R ₅ , (CR ₈ R ₈ ) _q C (O) NR ₄ R ₅ , R ₂ _iQanKeHMnC (O) NR ₄ R 5, (CR ₈ R ₈ ) _q C ( O) NR ₄ R ₁₀ , S (O) ₃ R ₈ , (CRgR ^ qCiOJR ^, R _{2. 10} alkenyl C (O) R _{11 (} R ₂ _io alkenylC (O) OR _{11 (} (CRgR ^ CiOJORn, (CRgRg ^ OC ^ Rn, (CR ₈ R ₈ ) _q NR ₄ C (O) R ₁₁ , (CR ₈ R ₈ ) _q C (NR ₄ ) NR ₄ R ₅ , (CRgR ^ NR.CiNRs) ^, (CR ₈ R ₈ ) _q NHS (O) ₂ R ₁₃ and (CR ₈ R ₈ ) _q S (O) ₂ NR ₄ R ₅ or two R} groups together form an O-CH 2 O- or 5 or 6 membered saturated or unsaturated ring and alkyl, aryl, aralkyl, heteroaryl, heterocyclyl groups may be optionally substituted; R ₄ and R ₅ are independently hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylC _b4 alkyl, optionally substituted heteroaryl, optionally substituted heteroarilC _V4 alkyl, heterocyclyl, heterocyclyl _b4 alkyl or R ₄ and R ₅ together with the nitrogen to which are attached form a 5 or 7 membered ring, optionally containing an additional heteroatom selected from oxygen, nitrogen or sulfur; R ₆ and R ₇ are independently hydrogen, C 1-6 alkyl, heteroaryl, aryl, alkylaryl and alkylC 1-6 heteroalkyl or R ₆ and R ₇ together with the nitrogen to which they are attached form a 5 or 7 membered ring which optionally contains an additional heteroatom which is selected from oxygen, nitrogen or sulfur and the ring may be optionally substituted; YeCR ₁₄ R ₁₅ , NR ₁₄ , 0, COnS (O) _t - *Yes R ₈ is hydrogen or C 1-6 alkyl; R ₉ is C _1-4 alkyl; R ₁₀ is C 1-6 alkyl C (O) ₂ R ₈ , Rn is hydrogen, optionally substituted C _b4 alkyl, optionally substituted aryl, optionally substituted arylC ^ alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC _1h alkyl, optionally substituted heterocyclyl, and optionally substituted heterocyclyl-alkyl; R ₁₂ is hydrogen, C _1-4 alkyl, aryl, arylC _1-4 alkyl, heteroaryl, heteroaryl Cf. ₄ alkyl, heterocyclyl and heterocyclylC 1-4 alkyl; R ₁₃ is C _b4 alkyl, aryl, arylC ^ alkyl, heteroaryl, heteroarylC _1h alkyl, heterocyclyl and heterocyclylC _{1. 4} alkyl; R ₁₄ and R ₁₅ are independently hydrogen, optionally substituted C 1-6 alkyl, OR _a and NR ₄ R ₅ ; or R ₁₄ and R ₁₅ together with the atom (s) to which they are attached may form a 4 to 7 membered ring which may optionally: contain an additional heteroatom, which heteroatom is selected from the group oxygen, nitrogen and sulfur, wherein the ring may optionally substituted; or a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein Rf is substituted at the 4-position by an electron-withdrawing moiety. The compound of claim 2, wherein Rf is a halogen, cyano or nitro group. The compound of claim 3, wherein Rf is halogen. The compound of claim 4, wherein Rf is independently fluorine, chlorine or bromine. The compound of claim 1, wherein R _b is hydrogen, C _1-4 alkyl or C ₁ . ₄ alkyl substituted with C (O) OH or C (O) OR _a . A compound according to claim 1, which is selected from the group: N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 M'cyclohexylurea; N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N'-1-adamantyl) urea; N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 N '- (tetrahydro-2-pyranyl) urea; N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '- (3-tetrahydrofuryl) urea; 6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) ureido] benzenesulfonamide; N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -1'-cyclohexylurea; 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethyl-cyclopropyl) ureido] benzenesulfonamide; 6-chloro-2-hydroxy-3- (3-piperidin-4-ylureido) benzenesulfonamide; 1- {1- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '- (4-methylcyclohexyl) urea; 6-chloro-2-hydroxy-3- [3- (3-methoxy-cyclohexyl) ureido] benzenesulfonamide; N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -1'-cyclopentylurea; 1 \ 1- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N'-cyclobutylurea; N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N'-cyclopropylurea; N- [6-Chloro-3- (3-cyclopentyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid butyl ester; 1- [4-chloro-2-hydroxy-3- (piperazine-1-sulfonyl) -phenyl] -3-cyclopentyl-urea; '' *** N- [6-Chloro-3- (3-cyclobutyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid tert-butyl ester; 3- [3 - ((1S, 2S) -2-benzyloxy-cyclohexyl) -ureido] -6-chloro-2-hydroxy-benzenesulfonamide and 6-chloro-3- (3-cyclobutyl-ureido) -2-hydroxy-1,1'-N-dimethylbenzenesulfonamide. A compound according to claim 7 which is N- (3aminosulfonyl-4-chloro-2-hydroxyphenyl) -1 N'-cyclohexylurea. The compound of claim 7 which is in the form of its sodium salt. A compound according to claim 7 which is in the form of its potassium salt. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 and a pharmaceutically acceptable carrier or diluent. Use of a compound according to claim 1 for the preparation of a pharmaceutical composition for the treatment of chemokine mediated diseases in which the chemokine is bound to the IL-a or b receptor in a mammal. Use according to claim 12, wherein the chemokine mediated disease is from the group consisting of psoriasis, atopic dermatitis, osteoarthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, respiratory distress syndrome, inflammatory disease , ulcerative colitis, stroke, septic shock, multiple sclerosis, endotoxic shock, gram-negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft rejection, b Alzheimer's oil, allograft rejection, malaria, restenosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis and unwanted hematopoietic stem cell release and diseases caused by respiratory viruses, herpes viruses and hepatitis dysfunction, trauma, tension, tension, contusion, psoriatic arthritis, herpes, encephalitis, vasculitis of the central nervous system, traumatic brain damage, tumors of the central nervous system, subarachnois Alen hemorrhage, trauma after surgery, interstitial pneumonitis, hypersensitivity, crystalline arthritis, acute and chronic pancreatitis, acute alcoholic hepatitis, necrotizing enterocolitis, chronic sinusitis, uveitis, polymiosititis, polymiositis, , glossitis, airway obstruction, airway hyperreactivity, obliterating bronchitis organizing pneumonia, bronchiectasis, obliterating bronchiolitis, chronic bronchitis, cor pulmonae, dyspnoea, emphysema, hy erkapneya, hyperinflation, hypoxemia, hyperoxia-induced inflammations, hypoxia, surgical reduction of the volume of the lung, pulmonary fibrosis, pulmonary hypertension, right ventricular hipetrofiya, sarcoidosis, Crohn's small airways fan perfusion mismatching, wheeze, colds and lupus.