SK13282002A3 - IL-8 receptor antagonists - Google Patents

Abstract

This invention relates to novel compounds of Formula (I) to (VII), and compositions thereof, useful in the treatment of disease states mediated by the chemokine, Interleukin-8 (IL-8).

Description

Technical field

I

The present invention relates to novel sulfonamide substituted diphenylurea compounds, to pharmaceutical compositions comprising said compounds, to processes for their preparation and to their use in the treatment of diseases mediated by IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 and ENA-78.

BACKGROUND OF THE INVENTION

Many different names have been used to refer to interleukin 8 (IL-8) such as neutrophil attractant / activating protein-1 (NAP-1), monocyte-derived neutrophil chemotaxis factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte The chemotaxis factor, Interleukin-8, is a chemoattractant for neutrophils, basophils and a subset of T cells. It is produced by most nuclear cells, including macrophages, fibroblasts, endothelial and epithelial cells exposed to TBF, IL-1α, IL-1β or LPS, and neutrophils themselves when exposed to LPS or chemotaxis 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. et al., Science 243, 1467 (1989) and J. Biol. Chem., 264, 10621 (1989); Cassatella et al., J. Immunol. 148, 3216 (1992)).

GRP-α, GRO-β, GRO-γ, and NAP-2 also belong to the chemokine family. Like IL-8, these chemokines are referred to by different names. For example, GRO-α, -β, -γ are referred to as MGSAa, β. and γ (Melanoma Growth Stimulating Activity (MGSA)), see Richmond et al., J. Cell. Physiology 129, 375 (1986) and Chang et al., J. Immunol. 148: 451 (1992). All chemokines of the group and which contain the RLR motif directly preceding the CXC motif bind to the IL-8 B receptor (CXCR2).

IL-8, GRP-α, GRO-β, GRO-γ, NAP-2 and ENA-78 stimulate many functions in vitro. All of them have been shown to have chemoattractant properties towards neutrophils,

Whereas IL-8 and GRPa have T-cell and basophil chemotaxis activity. In addition, IL-8 may induce histamine release from basophils in normal and atopic individuals. GROα and IL-8 can further induce lysosomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase surface expression of Mac-1 (CD11b / CD18) on neutrophils without de novo protein synthesis. This may contribute to increased neutrophil adhesion to vascular endothelial cells. Many known diseases are characterized by mass infiltration of neutrophils. Since IL-8, GRPα, GROβ, GROγ and NAP-2 stimulate neutrophil accumulation and activation, they are involved in many acute and chronic inflammatory disorders including psoriasis and rheumatoid arthritis (Baggiolini et al., FEBS Lett. 307, 97 (1992); Miller et al. 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. Donnely et al., Lancet, 341, 643 (1993) In addition, ELR chemokines (those that contain the amino acid ELR cluster just prior to the CXC cluster) are also involved. in angiostasis (Strieter et al., Science 258, 1798 (1992)).

IL-8, GRP-α, GRO-β, GRO-γ and NAP-2 in vitro induce neutrophil shape change, chemotaxis, granule release, respiratory burst through binding to seventh-transmembrane receptors and their activation by a G-protein-binding moiety, in particular binding to IL-8 receptors and most preferably to the IL-δβ receptor (CXCR2) (Thomas et al., J. Biol. Chem. 266,14839 (1991); Holmes et al., Science 253, 1278 (1991)). An example for members of this receptor family is the development of non-peptide low molecular weight antagonists. For a review, see: Friedinger R., Progress in Drug Research, Vol. 40, pages 33-98, Birkhauser Verlag, Basel 1993.

The IL-8 receptor is therefore a promising target for the development of novel anti-inflammatory agents.

Two high affinity human IL-8 receptors (77% homology) are described: the IL-8Rα receptor, which binds only IL-8 with high affinity, and the second Ιί-8Ρβ, which has a high affinity for IL-8 but also for IL-8. GRP-α, GRO-β, GRO-γ, and NAP-2. (See Holmes et al., Supra; Murphy et al., Science 253, 1280 (1991); Lee et al., J. Biol.

-3 Chem., 267, 16283 (1992); La Rosa et al., J. Biol. Chem. 267, 25402 (1992); and Gayle et al., J. Biol. Chem. 268, 7283 (1993)).

In this field of treatment, there remains interest in compounds that are capable of binding to the IL-8 and / or β receptor. Therefore, compounds associated with IL-8 receptor binding inhibitors would be beneficial for conditions associated with increased IL-8 production (which is responsible for chemotaxis of neutrophil and T cell subgroups to the site of inflammation).

SUMMARY OF THE INVENTION

The invention relates to a method of treating a chemokine-mediated disease, wherein said chemokine is a chemokine that binds to the IL-8 α or β receptor and said method comprises administering an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof. In particular, said chemokine is IL-8.

The invention also relates to a method of inhibiting the binding of IL-8 to its receptors in a mammal in need thereof; the method comprising administering to the mammal in need of such treatment an effective amount of a compound of Formula I.

The invention relates to sulfonamide substituted diphenylurea compounds of formula I and to pharmaceutical compositions comprising a compound of formula I and a pharmaceutical carrier or diluent.

The present invention provides sulfonamide substituted diphenylurea compounds of formula I

where

R ^b is independently selected from the group consisting of hydrogen, NR ⁶ R ⁷ , OH, OR ^a , C 1 -C 5 alkyl, aryl, aryl-C 1 -C ₄ -alkyl, aryl-C 2 -C ₄ -alkenyl, cycloalkyl, cycloalkyl- CPCS-alkyl,

-4heteroaryl, heteroaryl-C ₁ -C ₄ -alkyl, heteroaryl-C ₂ -C ₄ -alkenyl, hetero ring, hetero ring C ₁ -C ₄ -alkyl and heter ring C ₂ -C ₄ -alkenyl; each of said groups may optionally be one to three times independently substituted with a substituent selected from the group consisting of halogen, nitro, halogen substituted C 1 -C ₄ -alkyl, C ₁ -C ₄ -alkyl, amino, mono- or di-C 1 -C ₄ -alkyl substituted amine, OR ^a , C (O) R ^a , NR ^and 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 ⁷ , 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 taken together form 3 to 10 a membered ring, optionally substituted and containing, in addition to carbon, independently, 1 to 3 groups selected from the group consisting of NR ^a , O, S, SO or SO 2; and wherein the substituent optionally may be unsaturated;

R ^a is selected from the group consisting of alkyl, aryl, aryl-C 1 -C 4 -alkyl, heteroaryl, heteroaryl-C 1 -C 4 -alkyl, hetero ring, COOR ^13, and hetero ring C 1 -C 4 -alkyl; each of said groups may be optionally substituted;

m is an integer of 1 to 3;

m 'is 0 or an integer of 1 or 2;

n is an integer from 1 to 3;

q is 0 or an integer from 1 to 10;

t is 0 or an integer of 1 or 2;

s is an integer of 1 to 3;

R ¹ is independently selected from the group consisting of hydrogen, halogen, nitro, cyano, C 1 -C 10 -alkyl, halogen substituted C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 1 -C 10 -alkoxy, halogen substituted C 1 -C 10 -alkoxy, azide, S (O) t R ^4, (CR ⁸ R ⁸⁾ q S (O) _t R ^4, hydroxy, hydroxy substituted C 4 alkyl, aryl, aryl-C4-alkyl, aryl-C2-Ci0alkenyl, aryloxy, aryl-Ci-C4- alkoxy, heteroaryl, heteroarylalkyl, heteroaryl-C 2 -C 10 alkenyl, heteroaryl-C 1 -C 4 -alkyloxy, hetero ring, hetero ring C 1 -C 4 -alkyl, hetero ring C 1 -C 4 alkyloxy, hetero ring C 2 -C 10 -alkenyl, (CR ⁸ R ⁸ ) q NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C (O) NR ⁴ R ⁵ , C 2 -C ₁₀ -alkenyl-C (O) NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C (O) NR ⁴ R ¹⁰ , S (O) 3R ⁸ , (CR ⁸ R ⁸ ) _q C (O) R ¹¹ ,

C ₂ -C 10 -alkenyl-C (O) R ¹¹ , C 2 -C 10 -alkenyl-C (O) OR ¹¹ , (CR ⁸ R ⁸ ) q C (O) OR ¹¹ , (CR ⁸ R ⁸ ) qOC (O) R ¹¹ , (CR ⁸ R ⁸ ) q NR 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 and (CR ⁸ R ⁸⁾ q S (O) ₂ NR ⁴ R ^5; or two R ¹ groups together may form O- (CH ₂ ) _S O- or a 5- to 6-membered saturated or unsaturated ring, and

The above-mentioned alkyl, aryl, arylalkyl, heteroaryl, heterocyclic groups may be optionally substituted;

R ⁴ and R ⁵ are independently from each other selected from the group consisting of hydrogen, optionally substituted C 1 -C ₄ -alkyl, optionally substituted aryl, optionally substituted arylC 1 -C ₄ -alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC 1 -C ₄ -alkyl , heterocyclic ring and heterocyclic ring-C 1 -C ₄ -alkyl; or R ⁴ and R ⁵ together with the nitrogen to which they are attached form a five to seven membered ring, which may optionally contain another heteroatom selected from O, N and S;

R ⁶ and R ⁷ are independently selected from the group consisting of hydrogen, C 1 -C ₄ -alkyl, heteroaryl, aryl, alkyl-aryl and alkyl-C 1 -C ₄ -heteroalkyl; or R ⁶ and R ⁷ together with the nitrogen to which they are attached form a 5- to 7-membered ring, which may optionally contain another heteroatom selected from O, N and S; wherein said ring may be optionally substituted;

Y is selected from the group consisting of CR ¹⁴ R ¹⁵ , NR ¹⁴ , O, CO and S (O) _t ;

R ⁸ is hydrogen or C 1 -C ₄ -alkyl;

R ⁹ is C 1 -C ₄ -alkyl;

R ¹⁰ is C 1 -C 10 -alkyl-C (O) ₂ R ⁸ ;

R ¹¹ is selected from the group consisting of hydrogen, optionally substituted C 1 -C ₄ -alkyl, optionally substituted aryl, optionally substituted aryl-C ₁ -C ₄ -alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-C 1 -C ₄ -alkyl, heterocyclic ring and optionally substituted heterocyclic-C ₁ -C ₄ -alkyl; and

R ¹² is selected from the group consisting of C 1 -C ₄ -alkyl, aryl, aryl-C ₁ -C ₄ -alkyl, heteroaryl, heteroaryl-C 1 -C ₄ -alkyl, a heterocyclic ring and a heterocyclic ring-C ₁ -C ₄ -alkyl;

R ¹³ is selected from the group consisting of C ₁ -C ₄ -alkyl, aryl, aryl-C ₁ -C ₄ -alkyl, heteroaryl, heteroaryl-C ₁ -C ₄ -alkyl, a heterocyclic ring and a heterocyclic ring-C 1 -C ₄ -alkyl;

R ¹⁴ and R ¹⁵ are independently selected from the group consisting of hydrogen, optionally substituted C ₁ -C ₄ -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 four to seven membered ring which may optionally contain another heteroatom selected from the group consisting of oxygen, nitrogen and sulfur; wherein said ring may be optionally substituted;

-6 or a pharmaceutically acceptable salt thereof.

The compounds of Formula I may also be used in conjunction with veterinary treatment of non-human mammals in cases where inhibition of IL-8 or other chemokines that bind to IL-8 α and β receptors is required. Treatment of chemokine-mediated diseases, therapeutic or prophylactic, in animals includes the disease states listed in the Methods of Treatment section.

Preferably R ^b is independently hydrogen, NR ⁶ R ⁷ , OH, OR ^a , C 1 -C 4 -alkyl, aryl, aryl-C 1 -C 4 -alkyl, aryl-C 2 -C 4 -alkenyl, heteroaryl, heteroaryl-C 1 -C 4 -alkyl, heteroarylC2-C4-alkenyl, heterocycle, heterocycle-C1-C4-alkyl or heterocycle-C2-C4-alkenyl; each of said groups may optionally be one to three times independently substituted with halogen, nitro, halogen substituted with C 1 -C 4 -alkyl, C 1 -C 4 -alkyl, amino, mono- or di-C 1 -C 4 -alkyl substituted with amine, cycloalkyl, cycloalkyl-C 1 -C 5 -alkyl, OR ^a , C (O) R ^a , NR ^a C (O) OR ^a , OC (O) NR ⁶ R ⁷ , aryloxy, aryl-C 1 -C 4 -alkyloxy, hydroxy, C 1 -C 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) 2NR ⁶ R ⁷ , NHS ( O) ₂ R ^a .

Alternatively, the two mentioned R ^b substituents may together form a 3 to 10 membered ring, optionally substituted and containing in addition to carbon, independently 1 to 3 NR ⁹ , O, S, SO or SO ₂ groups, which may be optionally substituted.

Preferably R ^a is an alkyl, aryl, aryl-C 1 -C ₄ -alkyl, heteroaryl, heteroarylC ₁ -C ₄ -alkyl, hetero ring or a heterocycle-C ₁ -C ₄ -alkyl group; each of said groups may be optionally substituted.

Preferably R ¹ is independently selected from the group consisting of hydrogen, halogen, nitro, cyano, halogen substituted C 1 -C 10 -alkyl such as CF 3, C 1 -C 10 alkyl such as methyl, ethyl, isopropyl or n-propyl, C2 -C 10 -alkenyl, C 1 -C 10 alkoxy, such as methoxy or ethoxy, halogen substituted C 1 -C 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 -C 4 -alkyl such as methanol or ethanol, aryl such as phenyl or naphthyl, aryl-C 1 -C 4 -alkyl such as benzyl, aryloxy such as phenoxy, aryl-C 1 -C 4 -alkoxy such as benzyloxy; heteroaryl, heteroarylalkyl, heteroaryl-C, -C4-alkyloxy, aryl-C2-Cw alkenyl, heteroaryl C2-C10 alkenyl, heterocyclic C2-Ci0 alkenyl, (CR ⁸ R ⁸⁾ q NR ⁴ R ^5, C 2 -C 10 alkenyl -C (O) NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C (O) NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C (O) NR ⁴ R ¹⁰ , S (O) 3 H, S (O) ₃ R ⁸ , (CR ⁸ R ⁸ ) _q C (O) R ¹¹ ,

-7C ₂ -C 10 -alkenyl-C (O) R ¹¹ , C 2 -C 10 -alkenyl-C (O) OR ¹¹ , (CR ⁸ R ⁸ ) q C (O) R ¹¹ , (CR ⁸ R ⁸ ) _q C ( O) OR ¹¹ , (CR ⁸ R ⁸ ) qOC (O) R ¹¹ , (CR ⁸ R ⁸ ) q NR 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 and (CR ⁸ R ⁸ ) q S (O) ₂ NR ⁴ R ⁵ . All groups containing aryl, heteroaryl and heterocyclic groups may be optionally substituted as follows. ¹

The term aryl, heteroaryl and heterocyclic groups refers herein to alkyl and, when included, alkenyl rings such as aryl, arylalkyl and arylalkenyl rings. The term groups and circles may be used interchangeably herein.

Preferably R ⁴ and R ⁵ are independently from each other hydrogen, optionally substituted C 1 -C 4 alkyl, optionally substituted aryl, optionally substituted aryl-C 1 -C 4 alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-C 1 -C 4 -alkyl, heterocyclic ring ^: and heterocyclic ring- C, -C4-alkyl; or R ⁴ and R ⁵ together with the nitrogen to which they are attached form a five to seven membered ring, which may optionally contain another heteroatom selected from O, N and S;

Preferably R ⁶ and R ⁷ are each independently hydrogen, C _r C ₄ -alkyl, and heteroaryl _-alkyl--C4 heteroalkyl; or R ⁶ and R ⁷ together with the nitrogen to which they are attached form a five to seven-membered ring, which may optionally contain another heteroatom selected from oxygen, nitrogen and sulfur; wherein said ring may be optionally substituted;

Preferably R ⁸ is independently hydrogen or C 1 -C ₄ -alkyl;

Preferably R ⁹ is hydrogen or C ₁ -C ₄ -alkyl;

Preferably q is 0 or an integer having a value of 1 to 10.

Preferably, R ¹⁰ is -C-alkyl-Cio ^ R ⁸ as _CH2 C (O) ₂ H or _CH2 C (O ₂₎ CH 3;

Preferably R ¹¹ is hydrogen, C ₁ -C ₄ -alkyl, aryl, aryl-C 1 -C ₄ -alkyl, heteroaryl, heteroaryl-C 1 -C ₄ -alkyl, a heterocyclic ring and a heterocyclic ring-C 1 -C ₄ -alkyl.

Preferably R ¹² is hydrogen, C 1 -C 10 -alkyl, aryl, arylalkyl.

Preferably R ¹³ is C 1 -C ₄ -alkyl, aryl, arylalkyl, heteroaryl, heteroaryl-C 1 -C ₄ -alkyl, a heterocyclic ring and a heterocyclic ring-C 1 -C ₄ -alkyl, wherein all groups containing aryl, heteroaryl and the hetero ring may be optionally substituted.

Preferably R ¹⁴ and R ¹⁵ are hydrogen, optionally substituted C 1 -C 4 -alkyl, OR ^a and NR ⁴ R ⁵ ; or R ¹⁴ and R ¹⁵ together with the atom (s) to which they are attached

-8 may form a four to seven membered ring, which may optionally contain an additional heteroatom selected from the group consisting of oxygen, nitrogen and sulfur; wherein said ring may be optionally substituted.

Preferably Y is CR ¹⁴ R ¹⁵ , NR ¹⁴ O.

Preferably R ^a is alkyl, aryl-C 1 -C 4 -alkyl, heteroaryl, heteroaryl-C 1 -C ₄ -alkyl, a heterocycle or a heterocycle-C 1 -C ₄ -alkyl, each of which may be substituted.

Unless otherwise indicated, the term optionally substituted herein means groups such as halogen, for example, fluorine, chlorine, bromine or iodine, hydroxy; hydroxy substituted C _r Cio-alkyl, C _r Cio-alkoxy, such as methoxy or ethoxy, S (0) m'Ci-Cio-alkyl, wherein m 'is 0, 1, or 2, such as methyl thio, methylsulfinyl or methyl sulfonyl ; amino, mono- and di-substituted amino as in NR ⁸ R ¹² , NHC (O) R ¹³ , C (O) NR ⁸ R ¹² , C (O) OH, S (O) 2NR ⁸ R ¹² , NHS (O) 2 R ¹³ , C 1 -C 10 -alkyl such as methyl, ethyl, propyl, isopropyl or tert-butyl, halogen-substituted C 1 -C 10 -alkyl, for example CF 3, optionally substituted aryl such as phenyl, or optionally substituted arylalkyl such as benzyl or phenethyl, an optionally substituted heterocyclic ring, an optionally substituted heterocyclic alkyl, an optionally substituted heteroaryl, an optionally substituted heteroarylalkyl, wherein the aryl, heteroaryl and heterocyclic groups may be halogen substituted one to two times; hydroxy; hydroxy-substituted alkyl, C 1 -C 10 -alkoxy; S (0) Ci-Cio-alkyl; amino, mono- and di-substituted alkylamino as in NR ⁶ R ⁷ ; C 1 -C 10 -alkyl or halogen substituted C 1 -C 10 -alkyl, for example CF ₃ .

Suitable pharmaceutically acceptable salts are known to those skilled in the art and include basic salts of inorganic and organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, acetic acid, malic acid, tartaric acid, citric acid. , lactic acid, oxalic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid, phenylacetic acid and hydroxyphenylacetic acid. In addition, pharmaceutically acceptable salts of the compounds of Formula I may be formed with a pharmaceutically acceptable cation. Suitable pharmaceutically acceptable cations are those in U.S. Pat

The art is well known in the art and includes alkali metal cations, caustic earth metal cations, ammonium and quaternary ammonium cations.

The text uses terms that indicate:

- halogen - refers to all halogens, i.e. chlorine, fluorine, bromine and iodine; .

- C 1 -C ₁₀ -alkyl or alkyl - denote unbranched or branched chain groups having 1 to 10 carbon atoms, unless the chain length is otherwise limited, including but not limited to methyl, ethyl, n-propyl, iso -propyl, n butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl and the like;

cycloalkyl is used herein as a ring group, preferably having 3 to 8 carbon atoms, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl and the like;

- alkenyl is in all cases used herein as a straight or branched chain group having 2 to 12 carbon atoms, unless the chain length is otherwise limited, including but not limited to, ethenyl, 1-propenyl, 2-methyl-1-propenyl , 1-butenyl, 2-butenyl and the like;

- aryl - represents phenyl, naphthyl;

heteroaryl (alone or in combination such as heteroaryloxy or heteroarylalkyl) refers to a 5 to 10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, such as (but not limited to) not only) pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, tetrazole, thiazole, thiadiazole, triazole, imidazole or benzimidazole;

- heterocyclic ”or heterocycle (alone or in combination, such as heterocyclic alkyl) means a saturated or partially unsaturated four to ten membered ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur ; for example (but not limited to) pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, thiomorpholine or imidazolidine. Further, the sulfur may optionally be oxidized to a sulfone or sulfoxide;

the terms arylalkyl or heteroarylalkyl or heterocyclic alkyl are used herein to mean C 1 -C 10 -alkyl as defined above, attached to the aryl group,

A 10-heteroaryl or heterocyclic alkyl group as defined above, unless otherwise indicated individually;

sulfinyl-S (O) oxide of the respective sulfide, thio refers to sulfide, and sulfonyl refers to the totally oxidized S (O) 2 group _;

a phrase wherein two R ¹ groups together may form a five- or six-membered saturated or unsaturated ring is used herein to mean an aromatic ring system such as naphthalene, or is a phenyl group attached to a partially saturated or unsaturated ring such as C6- cycloalkenyl, for example by cyclohexene or C 5 -cycloalkenyl such as cyclopentene.

Representative compounds of formula I are, for example:

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

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

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

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

(5) 6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) -ureido] -benzenesulfonamide;

(6) N - [4-chloro-2-hydroxy-3-sulfamylphenyl] - N -cyclohexylurea;

(7) 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethylcyclopropyl) ureido] benzenesulfonamide;

(8) 6-chloro-2-hydroxy-3- (3-piperidin-4-yl-ureido) -benzenesulfonamide;

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

(10) 6-chloro-2-hydroxy-3- [3- (3-methoxy-cyclohexyl) -ureido] -benzenesulfonamide;

(11) N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -AT-cyclopentylurea;

(12) N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N-cyclobutylurea;

(13) N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N-cyclopropylurea;

(14) 4- [6-chloro-3- (3-cyclopentyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid tert-butyl ester;

(15) 1- [4-chloro-2-hydroxy-3- (piperazine-1-sulfonyl) -phenyl] -3-cyclopentyl-urea;

(16) 4- [6-chloro-3- (3-cyclobutyl-ureido) -2-hydroxy-benzenesulfonyl] -piperazine-1-carboxylic acid tert-butyl ester;

(17) 3- [3 - ((1S, 2S) -2-benzyloxycyclohexylureido] -6-chloro-2-hydroxybenzenesulfonamide; and (18) 6-chloro-3- (3-cyclobutyl-ureido) -2-hydroxy N /, / \ T-dimethyl-benzenesulfonamide.

-11 Preparation Methods

Compounds of formula I can be obtained using synthetic procedures, some of which are illustrated in the schemes below. The illustrated scheme is useful for preparation of compounds of formula I having different groups R, ^Rb and Y, which are formed by reaction of the optional substituents which are suitably protected to achieve compatibility with the reactions outlined herein. Subsequent deprotection then generally provides the compounds of the present invention. Once the urea nucleus has been formed, other compounds of Formula I may be prepared by conventional functional group synthesis techniques; these techniques are known in the art.

Scheme 1

a) i) NCS, AcOH, H ₂ O; ii) NR'RH, pyr; b) H ₂ SO ₄ , HNO ₃ ; c) NaOAc, 18-crown-6-ether; d) H ₂ SO ₄ , MeOH; e) Pd / C, H ₂ ; f) RCNO, DMF.

The desired 4-chloro- N - (3-sulfonamido-2-hydroxyphenyl) - N -cycloalkylurea can be prepared from commercially available 2,6-dichlorothiophenol by the procedure outlined in Scheme 1. The thiol can be oxidized to the corresponding sulfonyl halide using a halogenating agent. agents such as NCS, NBS, Cl ₂ or Br ₂ in the presence of a protic solvent such as water, acetic acid, or an alcohol, or combinations thereof. The yield can be increased if a quencher such as sodium or potassium acetate is used in the reaction mixture and the reaction is conducted at or below room temperature. The corresponding sulfonyl halide can then be condensed with an amine in the presence of a base such as pyridine, triethylamine, potassium carbonate or sodium hydride to provide the analogous sulfonamide 2 in Scheme 1. Said sulfonamide 2 in Scheme 1 can be nitrated under strong nitration conditions such as nitric acid in sulfuric acid. to form the aromatic nitro compound 3 in Scheme 1. The chlorine ortho-position to the nitro group can be selectively hydrolyzed using acetate such as sodium acetate in the presence of a crown ether such as 18 crown-6-ether to give acetate 4 in Scheme 1. Said acetate group can be hydrolyzed under acidic conditions in an alcoholic solvent such as methanol or ethanol with a catalytic amount of acid to give phenol 5 in Scheme 1. The nitro group can be reduced under conditions well known in the art, for example, hydrogen in the presence of palladium on carbon, stannous chloride in methanol, zinc vp Aniline can then be coupled with a commercially available isocyanate or thioisocyanate to give the desired urea or thiourea. Alternatively, the desired isocyanates may be prepared by condensing an amine with triphosgene in the presence of a base (such as potassium carbonate) or by reacting a carboxylic acid with diphenylphosphoryl azide in the presence of a base (such as triethylamine).

Scheme 2

13a) NaH, R'X; b) NaH, RX

If the sulfonamide 1 in Scheme 2 (3 in Scheme 1) is free of the functional groups R 1 = R = H, then the alkylation may be used to form the derivative required herein. Said sulfonamide is deprotonated with a base such as sodium hydride and then alkylated using an alkyl halide such as benzyl bromide or methyl iodide to form 2 in Scheme 2. The sulfonamide can then be alkylated a second time using sodium hydride and another alkyl halide to form 3 in Scheme 2. convert to the desired urea as described in Scheme 1.

Scheme 3

a) i) NCS, AcOH, H ₂ O; ii) NaOH, MeOH; b) H ₂ SO ₄ , HNO ₃ ; c) NaOH, MeOH; d) PCI ₅ , POCl ₃ ; e) NHR'R, Et ₃ N.

An alternative method of preparing compound 5 in Scheme 3 (3 in Scheme 1) is shown in Scheme 3 wherein the commercially available 2,6-dichlorothiol is oxidized to a 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. Said sulfonyl halide may be hydrolyzed using a metal hydroxide such as sodium hydroxide or potassium hydroxide to give the corresponding sulfonic acid salt. The sulfonic acid salt can then be nitrated using nitric acid in a solvent (a strong acid such as sulfuric acid) to form nitrophenylsulfonic acid.

The sulfonic acid 3 in Scheme 3 can be converted to the sulfonamide 5 in Scheme 3 using a three-step process that involves the formation of a metal salt with a base such as sodium hydroxide, sodium hydride, or sodium carbonate to form 4 in Scheme 3. the sulfonic acid salt is then converted to sulfonyl chloride using PCl5 with POCl 3 as solvent. The sulfonyl chloride can then be converted to the corresponding sulfonamide 5 in Scheme 3 (3 in Scheme 1) using the appropriate amine HNR'R in triethylamine at a temperature ranging from -78 to 60 ° C. The sulfonamide 5 in Scheme 3 can be further processed by the methods described in Scheme 1. This method is not limited to 2,6-dichlorothiol; 2,6-difluorothiol, 2,6-dibromothiol and 2,6-diiodothiol can also be used. The halogens in these compounds can be converted to the corresponding cyano, amino, thiol or alkoxy compounds by a nucleophilic exchange reaction using nucleophiles such as alkylthiolates, alkoxides, amines, and cyanides. Said halogens may be further replaced by palladium coupling and carbonylation, as is well known in the art, to provide the corresponding amido, carbonyl, alkenyl, alkyl, phenyl, and heterocyclic substituted products of formula (I).

Methods of treatment

The compounds of formula I, or pharmaceutically acceptable salts thereof, may be used for the preparation of medicaments for the prophylactic or therapeutic treatment of a human or other mammal which is in any disease state caused or exacerbated by excessive or unregulated production of IL-8 cytokines by mammalian cells such as not only) monocytes and / or macrophages, or other chemokines that bind to IL-8 and / or β receptors, also referred to as I. or II receptors. type.

Accordingly, the invention relates to a method of treating a chemokine-mediated disease, wherein the chemokine is one that binds to IL-8 and / or the β receptor; said method comprising administering an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof. In particular, said chemokines are IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 and ENA-78.

The compounds of formula I are administered in an amount sufficient to inhibit cytokine function, particularly IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 function.

- 15 or ENA-78 to return to normal levels of their physiological functions or, in some cases, to subnormal levels, by bio-regulation to improve the disease state. Abnormal levels of IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78, in the context of the present invention, mean: (a) levels of free IL-8 greater than or equal to 1 picogram in 1 ml; (b) any cells associated with IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78 above normal physiological levels; or (c) the presence of IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78 above basal levels in cells or tissues in which IL-8, GRO-α, GRO- β, GTO-γ, NAP-2, or ENA-78.

Compounds of formula I have generally been shown to have improved oral bioavailability and longer tv ₂ compared to those disclosed in WO 96/25 157 and WO 97/29 743; said patents are incorporated herein by reference.

Many disease states are known in which excessive or unregulated IL-8 production affects exacerbation and / or disease development. Chemokine-mediated diseases include: psoriasis, atopic dermatitis, osteoarthritis, rheumatoid arthrosis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septum, scar, stroke , toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, host versus graft reaction, Alzheimer's disease, allograft rejection, malaria, restenosis, angiogenesis, atherosclerosis, osteoporosis and non-gastrointestinal disease, gingivitis, gingivitis herpes viruses and hepatitis viruses, meningitis, cystic fibrosis, premature labor, cough, itching, multi-organ dysfunction, trauma, tension, repulsion, bruising, psoriatic arthritis, herpes, encephalitis itide, CNS vasculitis, traumatic brain injury, CNS tumors, subarachnoid haemorrhage, postoperative trauma, interstitial pneumonia, hypersensitivity, crystal-induced arthritis, acute and chronic pancreatitis, acute alcohol hepatitis, necrosis, necrotis, necrotis, necrotis, necrotis, necrotis, necrotis

-16vasculitis, acne, gastric and duodenal ulcers, celiac disease, oesophagitis, glositis, airway obstruction, respiratory hyperreaction, bronchiolitis, inflammatory pulmonary inflammation, bronchiectasis, bronchiolitis, bronchiolitis, hypernitus, bronchiolchitis, respiratory hyperchiolchitis, emphysema, respiratory palsy, emphysema, heart disease , hyperoxia-induced inflammation, hypoxia, lung surgery, pulmonary fibrosis, pulmonary hypertension, ventricular hypertropy, sarcoidosis, small airway disease, ventilation-perfusion disorders, wheezing, rhinitis and lupus.

These diseases are primarily characterized by massive neutrophil infiltration, T-cell infiltration or neovascular growth and are associated with increased production of IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78, which is responsible for neutrophil chemotaxis to the site of inflammation or directed endothelial cell growth. Unlike other inflammatory cytokines (IL-1, TNF and IL-6), the cytokines IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78 have a particular property of promoting neutrophil chemotaxis, release enzyme including, but not limited to, elastase release as well as production and activation of superoxide. The chemokines α, but especially the chemokines GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78 may, despite their effect on IL-8 receptors type I or II, promote neovascularization of tumors by promoting directed growth of endothelial cells. Thus, inhibition of IL-8 induced chemotaxis or activation may directly lead to a reduction in neutrophil infiltration.

Recent evidence confirms the involvement and role of chemokines in the treatment of HIV infections; Littleman et al., Nature 381, 661 (1996) and Koup et al., Nature 381, 667 (1996).

Recently, reference has also been made to the use of IL-8 inhibitors in the treatment of atherosclerosis. The first reference (Boisvert et al., J. Clin. Invest. 101: 353-363 (1998)) shows that in the bone marrow transplantation of a mouse deficient in LDL receptors, the absence of IL-8 receptors on stem cells (and therefore on monocytes). (macrophages) results in decreased development of atherosclerotic plaques. Other supporting links are: Apostopoulus et al., Arterioscler. Thromb. Vasc. Biol. 16, 1007-1012 (1996); Liu et al., Arterioscler. Thromb. Vasc. Biol. 17: 317-323 (1997); Rus et al., Atherosclerosis 127: 263-271 (1996); Wang et al., J. Biol. Chem. 271: 6637-8842 (1996); Yue et al., Eur. J. Pharmacol. 240, 81-84 (1993); Koch et al., Am. J. Pathol. 142: 1423-1431 (1993); Lee et al.

-17Immunol. Lett. 53, 109-113 (1996); and Terkeltaud et al., Arterioscler. Thromb. 14, 47-53 (1994)).

The invention further relates to a method of treatment, in acute conditions as well as in prevention, of subjects deemed susceptible to CNS damage; said method is based on the use of compounds of formula I which are chemokine receptor antagonists.

CNS lesions are intended to include open or penetrating head trauma, such as in head surgery, or closed head lesions, such as head trauma. Ischemic defeats are also within the scope of CNS damage, particularly in the brain.

Ischemic stroke can be understood as a focal neurological disorder that results in insufficient blood flow to a certain part of the brain, usually as a result of embolism, thrombus or local atheromatous occlusion of blood vessels. The role of inflammatory chemokines in this field has become increasingly important, and the present invention provides a means for the potential treatment of said disorders. A relatively inexpensive treatment is now available for the treatment of acute injury per se.

TNF-α is a pro-inflammatory cytokine, including expression of endothelial leukocyte adhesion molecules. Leukocytes infiltrate ischemic brain lesions and therefore compounds that inhibit or decrease TNF levels may be useful in the treatment of ischemic brain injury. See: Liu et al., Stroke 25 (7), 1481-1488 (1994); this disclosure is incorporated herein by reference.

Closed head injury models and treatment with mixed 5-LO / CO agents are discussed in Shohami et al., J. of Vaisc. Clinical Physiology and Pharmacology 3 (2): 99-107 (1992); this disclosure is incorporated herein by reference. In animals, it has been shown that treatment that reduces edema improves treatment outcome.

The compounds of formula I are administered in an amount sufficient to inhibit IL-8, to inhibit binding to IL-8 α or β receptors, as evidenced by decreased neutrophil chemotaxis and activation. The discovery that the compounds of formula i are inhibitors of IL-8 binding is based on the effects of said compounds of formula I as determined in the in vitro receptor binding studies reported herein. The compounds of formula I have been shown to be inhibitors of IL-8 type II receptors.

The term as used herein as an IL-8 mediated disease or condition refers to any of the disease states in which IL-8, GRO-α, GRO-β, GTO-γ, NAP-2, or ENA-78 either by producing IL8, GRO-α, GRO-β, GTO-γ, NAP-2 or ENA-78 alone, or when IL-8, GROα, GRO-β, GTO-γ, NAP-2 or ENA- 78 causes the release of monokines other than but not limited to IL-1, IL-6 and TNF. For example, a disease state in which IL-1 is a major component and whose production or effect is exacerbated or secreted in response to IL-8 is therefore considered to be an IL-8 mediated disease state.

The term chemokine-mediated disease or condition refers to any disease state in which a chemokine, such as IL-8, GRO-α, GRO-β, GTO-γ, NAP-2 is involved in binding to IL-8 and / or the β receptor. or ENA-78. Said diseases include conditions in which IL-8 is involved or by the production of IL-8 alone, or IL-8 causes the release of a chemokine other than (but not limited to) IL-1, IL-6 or TNF. A disease state in which, for example, it is a major component of IL-1 and whose production or effect is exacerbated or secreted in response to IL-8 is therefore considered a disease state mediated by IL-8.

The term cytokine herein refers to any polypeptide that affects cell functions and is a molecule that modulates interactions between cells in the immune, inflammatory, or hematopoietic response. Cytokine includes, but is not limited to, monokines and lymphokines, regardless of what cells produce them. For example, it is generally recognized that monokine is produced and secreted by a mononuclear cell such as a macrophage and / or monocyte. Monokines also produce many other cells such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epidermal keratinocytes and B-lymphocytes. It is generally reported that lymphokines are 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 refers herein to any polypeptide that affects cell functions and is a molecule that modulates interactions between cells in the cell.

- an immune, inflammatory, or hematopoietic response, similar to the term cytokine mentioned above. Chemokine is primarily secreted through cell transmembranes and causes chemotaxis and activation of specific white blood cells and leukocytes, neutrophils, monocytes, macrophages, T cells, B cells, endothelial cells and smooth muscle cells. Examples of chemokines include, but are not limited to, IL-8, GRO-α, GRO-β, GTO-γ, NAP-2, ENA-78, IP-10, MIP1α, β-β, PF4, and MCP1. , 2 and 3.

The compound of formula I or a pharmaceutically acceptable salt thereof is normally formulated into a pharmaceutical composition for use in therapy in accordance with standard pharmaceutical practice. The invention therefore 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) or pharmaceutically acceptable salts thereof and the pharmaceutical compositions containing said compounds or salts thereof may conveniently be administered by any of the methods commonly used for drug administration, for example, orally, topically, parenterally or by inhalation. Said compounds of formula (I) may be administered in conventional dosage forms prepared by conventional means by combining a compound of formula (I) with standard pharmaceutical carriers. The compounds of formula I may also be administered in conventional dosages in combination with a known, second therapeutically active compound. Methods of preparation include mixing, granulating and compressing, or dissolving suitable ingredients into the desired composition. It will be appreciated that 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 coupled, the route of administration, and other variables known in the art. The carrier (s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier used may be, for example, a solid or a liquid. Examples of solid carriers are lactose, white clay, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Examples of liquid carriers are syrup, peanut oil, olive oil, water and the like. The carrier or diluent may include a delayed or controlled agent

-20 release of the active substance; said substances are known in the art as, for example, gylceryl monostearate, distearate, alone or with a wax.

A wide variety of pharmaceutical formulations can be used. When a solid carrier is used, the preparation may be tabletted, placed in powder or pellet form in hard gelatin capsules, or in the form of powders or lozenges. The amount of solid carrier will be in a wide range, but is preferably from about 25 mg to about 1 gram. When a liquid carrier is used, the preparation will be in the form of a syrup, an emulsion, a soft gelatin capsule, a sterile injectable solution in an ampoule, or a non-aqueous liquid suspension.

The compounds of formula (I) may be administered topically, i.e. by non-systemic administration. This includes the use of a compound of formula I externally on the skin or buccal cavities and by dripping a compound of formula I into the ear, eye or nose, wherein said compound does not significantly pass into the bloodstream. On the other hand, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

Pharmaceutical compositions suitable for topical administration include semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as ointments, lotions, paste creams; and drops for administration to the eye, ear and nose. The active ingredient may be present in the topical preparation in an amount of from 0.001% to 10% by weight, for example from 1% to 2% by weight of the total weight of the pharmaceutical composition. The pharmaceutical composition may also contain 10% by weight, but preferably contains less than 5% by weight, more preferably from 0.1% to 1% by weight, based on the total weight of the pharmaceutical composition.

The waters of the present invention include those waters suitable for use in the skin or eye. The ophthalmic waters may consist of a sterile aqueous solution, optionally containing a bactericidal substance, and may be prepared by methods similar to drops. Waters or ointments for use on the skin may include a drying-accelerating component and causing the skin to cool, for example alcohol or acetone, and / or a moisturizer, for example glycerol or an oil such as castor oil or peanut oil.

The creams, ointments or pastes of the present invention are semi-solid formulations of the active ingredient for external use. They may be prepared by mixing the active ingredient in finely divided form or powder or as a solution or suspension in an aqueous or non-aqueous liquid, using a suitable device with an ointment or non-oily base. The base may consist of hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, metal soap; slime; a natural oil such as almond oil, corn oil, peanut oil, castor oil or olive oil; lanolin or derivatives thereof, or a fatty acid such as stearic acid or oleic acid together with an alcohol such as propylene glycol or macrogel. The pharmaceutical composition may suitably contain any suitable surfactant such as an anionic, cationic or nonionic surfactant, for example a sorbitan ester, or a polyoxyethylene derivative thereof. The pharmaceutical composition may further comprise substances which promote the formation and stability of dispersions and suspensions, such as natural gums, cellulose derivatives, or inorganic materials such as silicas and other additives, for example lanolin.

The drops of the invention may consist of sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal substance and / or a fungicidal substance and / or other suitable preservative and preferably comprise a surfactant. The resulting solution can then be clarified by filtration, transferred to a suitable container, which is sealed and sterilized by autoclaving or holding at 98-100 ° C for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to the container under aseptic conditions. Examples of droplet bactericidal and fungicidal agents include phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%). Suitable solvents for preparing solutions in oil include glycerol, dilute alcohol and propylene glycol.

The compounds of formula I may be administered parenterally, i.e., intravenously, intramuscularly, subcutaneously, intranasally, intrarectally, intravaginally, or intraperitoneally. Generally preferred forms of parenteral administration are subcutaneous and intramuscular administration. Suitable dosage forms for such administration may be prepared by conventional techniques. Compounds of formula I

It can also be administered by inhalation, which is intranasal and oral inhalation. Suitable dosage forms for such administrations, such as aerosol formulations or metered inhalation doses, may be prepared by known and conventional techniques.

In all described uses of the compounds of Formula I, the daily oral dosage regimen will preferably be from about 0.01 mg to about 80 mg per kilogram of body weight. The daily parenteral dosage regimen will be from about 0.001 mg to about 80 mg per kilogram of body weight. The daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four times, preferably two or three times daily. The daily inhalation dosage regimen will preferably be from about 0.01 mg to about 1 mg per kilogram of body weight per day. One of ordinary skill in the art will recognize that the optimum amount and schedule of administration of single doses of a compound of Formula I or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the patient's condition, form, mode and site of administration and such optimum can be determined by conventional techniques. One of ordinary skill in the art will also appreciate that the optimum treatment, i.e., the number of doses of compound of Formula I or a pharmaceutically acceptable salt thereof administered per day, can be determined by those of ordinary skill in the art by routine testing of the regimen.

The invention is further described by means of the following examples and biological examples, which are intended to illustrate the invention in more detail and are not intended to limit the scope of the invention in any way.

DETAILED DESCRIPTION OF THE INVENTION

All temperature data are reported in ° C, all solvents are of the highest purity available, and all reactions are carried out under anhydrous conditions under argon shielding unless otherwise noted.

In the Examples, all temperatures are in degrees Celsius. Mass spectra were obtained with a fast atom bombardment VG Zab mass spectrometer, unless otherwise indicated. ¹ H NMR (in further NMR) spectra were recorded on a 250 MHz Bruker AM 250 or AM 400 spectrometer. Multiple signal labeling: s = singlet, d = doublet, t = triplet, q = quartet, m = multiple

-23 signal and br = wide signal. Sat means saturated solution, eq. means the molar equivalent ratio of the reactant relative to the main reactant.

General procedure for the synthesis of 2-hydroxy-3-amino-6-chlorobenzenesulfonamide

a) 2,6-Dichlorobenzenesulfonyl chloride

To a mixture of 200 mL (in another mL) of acetic acid, water, and dichloromethane (3/1/4, v / v) was added 2,6-dichlorobenzenethiol (10.0 grams (in another g), 55.8 millimoles (in another mmol). N-chlorosuccinimide (37.28 g, 279 mmol) and potassium acetate (2.29 g, 27.9 mmol). The resulting mixture was stirred at 0 ° C, then warmed to room temperature and stirred overnight. The mixture was then diluted with 200 mL of dichloromethane, washed with water (3 x 100 mL). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated to give the desired product (11 g, 80%). ¹ H 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 at 0 ° C to 100 mL of pyridine while anhydrous ammonia gas was passed through the solution simultaneously (4 hours, 0 ° C). . The mixture was acidified to pH> 1 with a solution of HCl (c = _H, C 6 mol.dm ^'3), extracted with ethyl acetate. The combined organic layers were then dried (anhydrous 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 two hours, then 200 mL of water was added to form a precipitate. The mixture was filtered. The collected white solid was washed with water and dried in vacuo to give the desired product (7.17 g, 76%). ¹ H NMR (DMSO-d ₆ ): δ 8.25 (s, 2H), 8.20 (d, 1H), 7.92 (d, 1H).

24d) 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 ether (5.95 g, 22, 5 mmol) in 50 ml of dimethylsulfoxide was heated at 45 ° C for 7 days. The mixture was acidified with dilute hydrochloric acid (i = 1 mol.dm ³ ) and extracted with ethyl acetate. The organic layer was concentrated to give the crude product. The crude product was purified by column chromatography on silica gel using ethyl acetate / hexane / acetic acid (50/49/1, v / v) as eluent; the desired product was obtained (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 h. condenser. Then the solvent was evaporated. The residue was diluted with ethyl acetate and washed with water. The organic layer was then dried (anhydrous sodium sulfate) and concentrated to give the desired product (1.0 g, 68%). EI MS (m / z) 251.1, 253.2 (M +).

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 Pd / C (10%, 500 mg). The mixture was bubbled with hydrogen and then stirred at room temperature under a hydrogen gas pressure at room temperature for 4 hours. The mixture was then filtered through celite and the celite was washed with methanol. The solvent was evaporated to give the desired product (91% of theory). EI MS (m / z) 221.1, 223.1 (M +).

Example 1

Standard procedure for condensation of aniline with isocyanate

Preparation of N- (3-Aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -cyclohexylurea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (150 mg, 0.67 mmol) and cyclohexyl isocyanate (95 μΙ, 0.74 mmol) in 1 mL of N, N-dimethylformamide was stirred at room temperature for 20 h. . The mixture was diluted with ethyl acetate and washed with water to give the crude product. The crude product was purified by column chromatography on silica gel using ethyl acetate / hexane (50/50, v / v) as eluent. Subsequent recrystallization from acetone / hexane gave the desired product (121 mg, 52%). LC-MS (m / z) 348.0 (M < ^{+ >} ).

Example 2

Preparation of N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N- (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 N, N-dimethylformamide was stirred at RT for 20 h rooms. The mixture was then diluted with ethyl acetate and washed with water to give the crude product. The crude product was purified by column chromatography on silica gel using ethyl acetate / hexane (50/50, v / v) as eluent. Thus, the desired product was obtained (122 mg, 56%). LC-MS (m / z) 400.0 (M < ^{+ >} ).

Example 3

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

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (17 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 then diluted with ethyl acetate and washed with water to give the crude product. The crude product was purified by column chromatography on silica gel using ethyl acetate / hexane (60/40, v / v) as eluent. This was followed by separation by Gilson HPLC to give the desired product (28 mg, 10%). LC-MS (m / z) 350.2 (M < ^{+ >} ).

-26Example 4

Standard procedure for the synthesis of ureas by coupling of carboxylic acids with aniline

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

To a solution of tetrahydrofuran-3-carboxylic acid (0.093 mL, 1.0 mmol) in DMF (0.5 mL) was added DPPA (0.25 mL, 1.2 mmol) and TEA (0.25 mL, 1 mL). , 8 mmol) and the reaction mixture was heated to 70 ° C. After 18 hours, 3-amino-6-chloro-2-hydroxybenzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated at 70 ° C for an additional 18 hours. It was then cooled and diluted with water, extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude residue was purified by HPLC to give 18 mg (5% of th.) Of N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N '(3-tetrahydrofuryl) 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 DMF (0.5 mL) was added DPPA (0.25 mL, 1.2 mmol) and TEA (0.25 mL, 1.8 mL). mmol) and the reaction mixture was heated at 90 ° C for 18 hours. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated at 90 ° C for an additional 18 hours. It was then cooled and diluted with water, extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude residue was purified by HPLC to give 29 mg (11% of theory) of 6-chloro-2-hydroxy-3- [3- (2-methylcyclopropyl) -ureido] -benzenesulfonamide. LC-MS (m / z) 320 (M < ^{+ >} ).

Example 6

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

To a solution of 2,2,3,3-tetramethyl-cyclopropanecarboxylic acid (0.142 mL, 1.0 mmol) in DMF (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 80 ° C for 18 h. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (1.0 mmol) was added and the reaction was

was heated at 80 ° C for an additional 18 hours. It was then cooled and diluted with water, extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude residue was purified by HPLC to give 96 mg of 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethyl-cyclopropyl) 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 tert-butyl ester (0.23 g, 1.0 mmol) in DMF (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 for 18 h. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (1.0 mmol) was added and the reaction mixture was heated at 80 ° C for an additional 18 hours. It was then cooled and diluted with water, extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. 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 /\/-.[4-chloro-2-hydroxy-3-sulfamylphenyll]-/\/’-(4-methylcyclohexyl)urea

To a solution of 4-methyl-cyclohexanecarboxylic acid (0.141 mL) in DMF (0.5 mL) was added DPPA (0.215 mL) and TEA (0.139 mL) and the reaction mixture was heated to 80 ° C. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (0.150) was added.

g) and the reaction mixture was heated at 80 ° C for an additional 18 hours. It was then cooled and diluted with water, extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Crude residue

It was purified by HPLC to give 15 mg of N- [4-chloro-2-hydroxy-3-sulfamylphenyl] -N- (4-methylcyclohexyl) 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 DMF (0.5 mL) was added DPPA (0.25 mL) and TEA (0.25 mL) and the reaction mixture was heated to 70 ° C. After 18 hours, 3-amino-6-chloro-2-hydroxy-benzenesulfonamide (0.222 g) was added and the reaction mixture was heated at 70 ° C for an additional 18 hours. The reaction mixture was then cooled and diluted with water and extracted with ethyl acetate. The organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure. 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

A / - (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N / '- cyclopentylurea

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 of N, N-dimethylformamide was stirred at RT for 20 h. rooms. Purification by column chromatography on silica gel using ethyl acetate / hexane (20/80, v / v) as eluent followed by recrystallization from diethyl ether / hexane gave the desired product (130 mg, 43%). LC-MS (m / z) 334.2 (M < ^{+ >} ).

Example 11 N- (3-Aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -cyclobutylurea

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 of N, N-dimethylformamide was heated under argon for 2 hours at 80 ° C. The reaction mixture was then cooled to room temperature and 3-amino-6-chloro-292-hydroxybenzenesulfonamide (556 mg, 2.50 mmol) was added. The reaction mixture was stirred at room temperature for 20 hours. Purification by Gilson HPLC using acetonitrile / water as eluent (10/90 to 90/10, v / v, over 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-cyclopropanylurea

A mixture of cyclopropanecarboxylic acid (500 mg, 5.81 mmol) of diphenylphosphoryl azide (1.58 g, 5.81 mmol) and triethylamine (0.81 mL, 5.81 mmol) in 3 mL of N, N-dimethylformamide under a protective atmosphere of argon was heated at 80 ° C for 3 hours. The reaction mixture was then cooled to room temperature and 3-amino-6-chloro-2-hydroxybenzenesulfonamide (200 mg, 0.89 mmol) was added. The reaction mixture was stirred at room temperature for 20 hours. Purification by Gilson HPLC using acetonitrile / water as eluent (10/90 to 90/10, v / v, over 10 minutes) gave the desired product (74 mg, 27%). LC-MS (m / z) 306.2 (M < ^{+ >} ).

Example 13 N- (3- (N-Boc-piperazine) -aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -cyclopentylurea

A solution of 3- N - (Boc-piperazine) amino-6-chloro-2-hydroxybenzenesulfonamide (200 mg, 0.51 mmol) and cyclopentyl isocyanate (57 μΙ, 0.5 mmol) in 2 mL of Ν-dimethylformamide was stirred for 20 hours at room temperature. Chromatographic purification on a silica gel column using ethyl acetate / hexane (30/70, v / v) as eluent followed by recrystallization from dichloromethane / hexane gave the desired product (180 mg, 70%).

LC-MS (m / z) 503.2 (M < ^{+ >} ).

-30Example 14

N- (3- (N-Boc-piperazine) aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclopentylurea hydrochloride

A solution of N - (3- N -Boc-piperazine) -aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -cyclopentylurea (140 mg, 0.27 mmol) in 2 mL of dilute hydrochloric acid (I want = 4 mol.dm ^'3 in 1,4-dioxane) was treated at room temperature. The mixture was then concentrated. Recrystallization from acetonitrile gave the desired product (105 mg, 88%). LC-MS (m / z) 403.2 (M < ^{+ >} ).

Example 15

A / - (3- (N / '- Boc-piperazine) aminosulfonyl-4-chloro-2-hydroxyphenyl) -N / -cyklobutylmočovina

A mixture of cyclobutanecarboxylic acid (500 mg, 4.99 mmol), diphenylphosphoryl azide (1.18 mL, 5.49 mmol) and triethylamine (0.76 mL, 5.49 mmol) in 3 mL of N, N-dimethylformamide was heated under argon under a protective atmosphere. 2 hours at 80 ° C. The reaction mixture was then cooled to room temperature and 3-N- (Bocpiperazine) amino-6-chloro-2-hydroxybenzenesulfonamide (1.95 g, 4.97 mmol) was added. The reaction mixture was stirred at room temperature for 20 hours. Chromatographic purification on a silica gel column using ethyl acetate / hexane (30/70, v / v) as the eluent gave 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 - [(1S, 2S) - (-) - 2-benzyloxycyclohexylurea (1S, 2S) - (-) - 2-benzyloxycyclohexylisocyanate

To a solution of di-tert-butyl dicarbonate (148 mg, 0.68 mmol) and 4-dimethylaminopyridine (87 mg, 0.49 mmol) in 5 mL of dichloromethane was added a solution of (1S, 2S) (-) - 2-benzyloxycyclohexylamine (100 mg, 0.49 mmol) in 5 mL of dichloromethane. mixture

The mixture was stirred at room temperature for 15 minutes, then concentrated to give the crude product. FT-IR: 2,237.92 cm ^-1 .

/ N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) - / V - [(1 S, 2 S) - (-) - 2-benzyloxy-cyclohexyl] -urea

A solution of 3-amino-6-chloro-2-hydroxybenzenesulfonamide (109 mg, 0.49 mmol) and crude (1S, 2S) - (-) - 2-benzyloxycyclohexylisocyanate in 2 mL of N, N-dimethylformamide were stirred for 20 hours at room temperature. Purification by Gilson HPLC using acetonitrile / water as eluent (10/90 to 90/10, v / v, over 10 minutes) gave the desired product (60 mg, 27% of theory over two steps). LC-MS (m / z) 454.0 (M < ^{+ >} ).

Example 17

Synthesis of N - (3- (N ^, -dimethyl) -aminosulfonyl-4-chloro-2-hydroxyphenyl) -Acyclobutylurea hydrochloride N - (3- (N-dimethyl) -aminosulfonyl-4-chloro-2-hydroxyphenyl) ) - / V-Boc / V-cyklobutylmočovina

A mixture of cyclobutylamine (14 μΙ, 0.16 mmol), trimethylaluminium (80 μΙ, 0.16 mmol) in 3 mL of dichloromethane was stirred at room temperature for 10 minutes under argon. 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 reaction mixture was stirred at room temperature for 20 hours. Chromatographic purification on a silica gel column using ethyl acetate / hexane (20/80, v / v) as eluent gave the desired product (36 mg, 51%). LC-MS (m / z) 448.2 (M < ^{+ >} ).

N - (3- (N-dimethyl) -aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -cyclobutylurea hydrochloride

A solution of N - (3- (N-dimethyl) aminosulfonyl-4-chloro-2-hydroxyphenyl) - N -Boc- N -cyclobutylurea (36 mg, 0.08 mmol) in 2 mL of dilute hydrochloric acid (c _H 2 (4 mol.dm ³ in dioxane) was stirred at room temperature for 1 hour. The mixture is then

-32skoncetrovala. Recrystallization from acetone / hexane gave the desired product (20 mg, 65%).

Elemental analysis:

theory: C 44.89, H 5.22, N 12.08;

experimental: 44.85% C, 5.00% H, 11.91% N.

Biological examples

Said IL-8 and GRO-α chemokine inhibitory effects of the compounds of this invention were determined by the following in vitro experiments:

Receptor Binding

[ ¹²⁵ L] IL-8 (human, recombinant) having a specific activity of 2000 Ci-mol ^-1 was obtained from Amersham Corp., Arlington Heights, IL. GRO-α was obtained from NEN (New England Nuclear). All other chemicals used were of analytical purity. High levels of recombinant human IL-8 type α and β receptors were individually expressed in ovarian cells by the Chinese hamster as described by Holmes (Holmes et al., Science 253, 1278 (1991)). Chinese hamster ovary membranes were homogenized according to a known protocol (Haour et al., J. Biol. Chem. 249: 2195-2205 (1974)) except that the homogenization buffer was replaced with 10 mM Tris-HCl, 1 mM MgSO4, 0.5 mM EDTA (ethylenediaminetetraacetic acid), 1 mM PMSF (α-toluenesulfonyl fluoride), 0.5 mg per liter leupeptin, pH 7.5. The membrane protein concentration was determined using the Pierce Co. kit. Micro-assay Kit using bovine serum albumin as standard. All analyzes were performed in a 96-well microplate format. Each reaction mixture contained ¹²⁵ l of IL-8 (0.25 nM) or ¹²⁵ l GRO-α and 0.5 pg.mľ ¹ of IL-8Ra or 1.0 pg.mľ ¹ of IL-8R membranes in 20 nriM bistrispropáne and 0.4 mM Tris HCl buffer, pH 8.0, containing 1.2 mM MgSO 4, 0.1 mM EDTA, 25 mM Na and 0.03% CHAPS. In addition, drug or test compound was added and previously dissolved in DMSO to achieve a final concentration between 0.01 nM and 100 μΜ. Analysis starts with the addition of ¹²⁵ L-IL-8. After an hour at room temperature, the plate was harvested using the apparatus

-33Tomtec 96-well Harvester for glass fiber filter cloth, blocked with 1% polyethyleneimine / 0.5% BSA, which was washed three times with 25 mM NaCl, 10 mM TrisHCl, 1 mM MgSO ₄ , 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. The filter was then dried and evaluated with a Betaplate Liquid Scintillation Counter. Said recombinant IL-8Rα or type I receptor is also referred to herein as intolerant receptor and recombinant IL-8Rβ or type II receptor is referred to as a tolerant receptor.

Representatives of the compounds of Formula I, Examples 1 to 18, had positive inhibitory potency at IC ₅₀ levels <30 μΜ in this assay.

Chemotaxic analysis

In vitro inhibitory properties of these compounds were determined by neutrophil chemotaxis analysis as described in: Current Protocol in Immunology, Vol. I, Suppl. I, Unit 6.12.2; this literature is incorporated herein by reference in its entirety. Neutrophils were isolated from human blood as described in Current Protocols in Immunology, Vol. I, Suppl. I, Unit 7.23.1 (the entire disclosure of which is incorporated herein by reference). IL-8, GRO-α, GRO-β, GRO-γ, and NAP-2 chemoattractants were loaded into the lower chamber of a 48-well well (Neuro Probe, Cabin John, MD, USA) at concentrations between 0.1 and 100 nM . The two chambers are separated by a polycarbonate filter (5 µm). When the compound of the invention is analyzed, it is mixed with the cells (0.001 to 1000 nM) just before the cells are added to the upper chamber. Incubation takes place between 45 and 90 minutes at 37 ° C in a humidified 5% CO 2 incubator. At the end of the incubation period, the polycarbonate membrane is removed and the upper side rinsed. The membrane is then stained according to the Diff Quick staining protocol (Baxter Products, McGaw Park, IL, USA). Cells that exhibited chemotaxis to chemokines were visually counted under a microscope. In general, four fields from each sample were counted, the results averaged to obtain the average number of cells that migrated. Each sample was analyzed in triplicate and each compound repeated at least four times. No compound was added to some cells (positive control cells), these cells represented the maximal chemotaxis response of the cells. In the case of a negative control (unstimulated) they were not added to the lower chamber

-34chemokíny. The difference between the positive control and the negative control indicates the chemotaxis activity of the cells.

Elastase release analysis

The compounds of this invention were tested for their ability to prevent the release of elastase from human neutrophils. Neutrophils are isolated from human blood as described in Current Protocols in Immunology, Vol. I, Suppl. 7.23.1. PMNs 0.88.10 ⁶ cells were dispersed in Ringer's solution (NaCl 118, KCl 4.56, NaHCO ₃ 25, KH ₂ PO ₄ 1.03, glucose 11.1, HEPES 5 mM, pH 7.4) and transferred to each well. wells (50 μΙ each) of a 96 well plate. Test compounds (0.001 to 1000 nM) were added to the plates in a volume of 50 μΙ, 50 μΙ Cytochalsin B (20 μg.mľ ¹ ) and Ringer's buffer (50 μΙ). Cells were allowed to warm to 37 ° C (5% CO 2, 95% relative humidity) over 5 minutes before addition of IL-8, GRO-α, GRO-β, GRO-γ, or NAP-2 to final concentrations of 0.01 to 1,000 nM. The reaction was allowed to proceed for 45 minutes and then the 96 well plate was centrifuged (800 xg, 5 minutes) and 100 μΙ of supernatant was collected. The supernatant was transferred 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 pg.mľ ^1, dissolved in phosphate-buffered saline solution. The plate was immediately placed in a fluorescence reader of 96 well plates (Cytofluor 2350, Millipore, Bedford, MA, USA) and data was read at 3 minute intervals as described by Nakajima et al., J. Biol. Chem. 254. 4027 (1979). The amount of elastase released from PMNs is calculated by measuring the rate of degradation of MeOSuc-Ala-Ala-Pro-Val-AMC.

Determination of TNF-α in brain injury by injury

This analysis serves to investigate the expression of tumor necrosis factor mRNA in specific regions of the brain; said analysis is performed experimentally by lateral percussion induced traumatic brain injury (TBI) of the rat. Adult Sprague-Dawley rats (n = 42) were used. The animals were anesthetized with sodium pentobarbital (60 mg.kg ^-1 ip) and were subjected to lateral percutaneous brain damage with medium fluid

-35 resistance (0.24 MPa) centered over the left temporaparietal cortex (n = 18) or undergoing sham treatment after anesthesia (n = 18). Animals were sacrificed by decapitation at 1, 6 and 24 hours after injury, brain was taken and specimens were prepared from left (damaged) parietal cortex (LC) tissue, corresponding to the contralateral right cortex (RC) area, from the surrounding parietal damaged area. cortex (LA), the respective adjacent region in the right cortex (RA), the left hippocampus (LH), and the right hippocampus (RH). Total RNA was isolated, Northern blot hybridization was performed and quantified relative to TNF-α positive RNA control (macrophage = 100%). It observed a significant increase in TNF-α RNA expression in LH (104 ± 17% positive control, p <0.05 vs sham), LC (105 ± 21%, p <0.05) and LA (69 ± 8%). , p <0.01) in the traumatized hemisphere an hour after injury. TNF-α RNA overexpression was also observed in LH (48 ± 8, p <0.05), LC (30 ± 3%, p <0.01) and LA (32 ± 3%, p <0.01) for 6 hours, which persisted after 24 hours. In the contralateral hemisphere, TNF-α mRNA expression was increased in RH (46 ± 2%, p <0.01), RC (4 ± 3%) and RA (22 ± 8%) one hour after injury and in RH (28 ± 11%) ), RC (7 ± 5%) and RA (26 ± 6%, p <0.05) at 6 hours but not 24 hours after injury. No changes in TNF-α mRNA expression were observed in any of the 6 brain regions in both the hemispheres and at all times in the sham group and in the untreated group. These results suggest that, after parasagital brain injury, the time dependence of TNF-α mRNA expression varies in specific areas of the brain, including those in the intact hemisphere. Since TNF-α is able to induce nerve growth factor (NGF) and stimulate the release of other cytokines from activated astrocytes, these post-traumatic changes in TNF-α gene expression are important in both the acute and regenerative responses to CNS trauma.

CNS damage - model for IL-1 β mRNA

This assay characterizes regional expression of interleukin 1β (IL-1β) mRNA in specific areas of the brain following experimental lateral fluid percussive traumatic brain injury (TBI) of the rat. Adults were used

-36 Sprague-Dawley rats (n = 42). The animals were anesthetized with sodium pentobarbital (60 mg.kg ^-1 ip) and subjected to lateral, perusual brain injury of moderate severity (0.24 MPa) centered on the left temporaparietal cortex (n = 18), or undergo surgery without damage after anesthesia ( sham treatment) (n = 18). Animals were sacrificed by decapitation at 1, 6 and 24 hours after injury, brain was taken and specimens were prepared from left (damaged) parietal cortex (LC) tissue, corresponding to the contralateral right cortex (RC) area, from the surrounding parietal damaged area. cortex (LA), the respective adjacent region in the right cortex (RA), the left hippocampus (LH), and the right hippocampus (RH). Total RNA was isolated, Northern blot hybridization was performed, and the amount of IL-β beta brain tissue was expressed as a percentage of the relative radioactivity from the IL-1β positive RNA macrophage that was run on the same gel. One hour after brain injury, a significant increase in IL-1 β mRNA expression in LC (20.0 ± 0.7% relative to positive control, n = 6, p <0.05 compared to sham animal), LH (24.5 ± 0.9%, p <0.05) and LA (21.5 ± 3.1%, p <0.05) in the damaged hemisphere and LH (5.0 ± 1.3%, p <0.05). No expression was observed in any of the brain regions in the sham and intact animals group. The results show that after TBI the time dependence of IL-β mRNA expression is regionally stimulated in specific regions of the brain. These regional cytokine changes such as IL-β are important in the post-traumatic regimen.

All publications cited herein, including but not limited to patents and patent applications, are incorporated herein by reference, unless the individual publications have been specifically incorporated herein before.

The above disclosure fully discloses a patent including preferred embodiments thereof. Modifications and improvements to the individual embodiments are within the scope of the following claims. It is believed that one skilled in the art can, without further elaboration using the foregoing description, utilize the present invention to its fullest extent. Therefore, the Examples are to be understood in particular as illustrative of the invention and in no way limit the scope thereof. Embodiments of the invention which are claimed as property or patent are set forth in the claims below.

7?

Sulfonamide substituted diphenylurea compounds of formula I

Claims (12)

PATENT Claims where R ^b is independently selected from the group consisting of hydrogen, NR ⁶ R ⁷ , OH, OR ^a , C 1 -C 5 alkyl, aryl, aryl-C 1 -C 4 alkyl, aryl-C 2 -C 4 -alkenyl, cycloalkyl, cycloalkyl-C 1 -C 5 -alkyl, heteroaryl, heteroaryl-C 1 -C 4 -alkyl, heteroaryl-C 2 -C 4 -alkenyl, hetero ring, hetero ring C 1 -C 4 -alkyl and hetero ring C 2 -C 4 -alkenyl; each of said groups may optionally be one to three times independently substituted with a substituent selected from the group consisting of halogen, nitro, halogen substituted C 1 -C 4 -alkyl, C 1 -C 4 -alkyl, amino, mono- or di-C 1 -C 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 ⁷ , C (O) OH, C (O) OR ^a , S (O) 2 N ⁶ R ⁷ , NHS (O) 2 R ^a , or the two R ^b substituents taken together form a 3 to 10 membered ring, optionally substituted and containing, in addition to carbon, independently, 1 to 3 groups selected from the group consisting of NR ^a , O, S, SO or SO ₂ ; and wherein the substituent optionally may be unsaturated; R ^a is selected from the group consisting of alkyl, aryl, aryl-C 1 -C 4 -alkyl, heteroaryl, heteroaryl-C 1 -C 4 -alkyl, hetero ring, COOR ^13, and hetero ring-C ₁ -C ₄ -alkyl; each of said groups may be optionally substituted; m is an integer of 1 to 3; m 'is 0 or an integer of 1 or 2; n is an integer from 1 to 3; q is 0 or an integer from 1 to 10; -38t is 0 or an integer of 1 or 2; s is an integer of 1 to 3; R ¹ is independently selected from the group consisting of hydrogen, halogen, nitro, cyano, C 1 -C 6 -alkyl, halogen substituted C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 1 -C 10 -alkoxy, halogen substituted C 1 -C 10 -alkoxy, azide, S (O) t R ⁴ , (CR ⁸ R ⁸ ) q S (O) _t R ⁴ , hydroxy, hydroxy substituted C 1 -C 4 -alkyl, aryl, aryl-C 1 -C 4 -alkyl, aryl-C 2 -C 10 alkenyl, aryloxy, aryl-C 1 -C 4 -alkoxy, heteroaryl, heteroarylalkyl, heteroaryl-C 2 -C 10 alkenyl, heteroaryl-C 1 -C 4 -alkyloxy, hetero ring, hetero ring C 1 -C 4 -alkyl, hetero ring C 1 -C 4 -alkyloxy, hetero ring C 2 -C 10 -alkenyl, (CR ⁸ R ⁸ ) q NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C (O) NR ⁴ R ⁵ , C 2 -C ₁₀ -alkenyl-C (O) NR ⁴ R ⁵ , (CR ⁸ R ⁸ ) q C ( O) NR ⁴ R ¹⁰ , S (O) 3 R ⁸ , (CR ⁸ R ⁸ ) _q C (O) R ¹¹ , C ₂ -C ₁₀ -alkenyl-C (O) R ¹¹ , C ₂ -C ₁₀ -alkenyl-C (O) OR ¹¹ , (CR ⁸ R ⁸ ) q C (O) OR ¹¹ , (CR ⁸ R ⁸ ) qOC (O ) R ¹¹ , (CR ⁸ R ⁸ ) q NR 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 and (CR ⁸ R ⁸⁾ q S (O) ₂ NR ⁴ R ^5; or two R ¹ groups together may form O- (CH ₂ ) _{S 0 -} or a 5- to 6-membered saturated or unsaturated ring, and wherein said alkyl, aryl, arylalkyl, heteroaryl, heterocyclic groups may be optionally substituted; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, optionally substituted -C ₄ -alkyl, optionally substituted aryl, optionally substituted arylCrC ₄ -alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl _r C ₄ -alkyl, a heterocyclic ring, and heterocycle-C _r C ₄ -alkyl; or R ⁴ and R ⁵ together with the nitrogen to which they are attached form a five to seven membered ring, which may optionally contain another heteroatom selected from O, N and S; R ⁶ and R ⁷ are independently selected from the group consisting of hydrogen, C ₁ -C ₄ -alkyl, heteroaryl, aryl, alkyl-aryl and alkyl-C 1 -C ₄ -heteroalkyl; or R ⁶ and R ⁷ together with the nitrogen to which they are attached form a 5- to 7-membered ring, which may optionally contain another heteroatom selected from O, N and S; wherein said ring may be optionally substituted; Y is selected from the group consisting of CR ¹⁴ R ¹⁵ , NR ¹⁴ , O, CO and S (O) _t ; R ⁸ is hydrogen or C ₁ -C ₄ -alkyl; R ⁹ is C ₁ -C ₄ -alkyl; R ¹⁰ is C 1 -C ₁₀ -alkyl-C (O) ₂ R ⁸ ; -39R ¹¹ is selected from the group consisting of hydrogen, optionally substituted C 4 -alkyl, optionally substituted aryl, optionally substituted _{aryl-Ci-C4-alkyl,} optionally substituted heteroaryl, optionally substituted heteroaryl-Ci-4 alkyl, heterocyclic a ring and optionally substituted heterocyclic-C ₁ -C ₄ -alkyl; and R ¹² is selected from the group consisting of C ₁ -C ₄ -alkyl, aryl, aryl-C ₁ -C ₄ -alkyl, heteroaryl, heteroaryl-C 1 -C ₄ -alkyl, a heterocyclic ring and a heterocyclic ring-C 1 -C ₄ -alkyl; R ¹³ is selected from the group consisting of C ₁ -C ₄ -alkyl, aryl, aryl-C 1 -C ₄ -alkyl, heteroaryl, heteroaryl-C ₁ -C ₄ -alkyl, heterocyclic ring and heterocyclic ring-C 1 -C ₄ alkyl; R ¹⁴ and R ¹⁵ are independently selected from the group consisting of hydrogen, optionally substituted C 1 -C ₄ -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 four to seven membered ring which may optionally contain another heteroatom selected from the group consisting of oxygen, nitrogen and sulfur; wherein said ring may be optionally substituted; or a pharmaceutically acceptable salt thereof. The sulfonamide substituted diphenylurea compounds of claim 1, wherein R ¹ is substituted at the 4-position with an electron withdrawing group. The sulfonamide substituted diphenylurea compounds according to claim 2, wherein R ¹ is halogen, cyano or nitro. The sulfonamide substituted diphenylurea compounds of claim 3, wherein R ¹ is halogen. The sulfonamide substituted diphenylurea compounds of claim 4, wherein R ¹ is independently fluoro, chloro or bromo. The sulfonamide substituted diphenylurea compounds of claim 1, wherein R ^b is hydrogen, C 1 -C ₄ -alkyl or C ₁ -C ₄ -alkyl substituted with C (O) OH or C (O) OR ^a . -407. The sulfonamide substituted diphenylurea compounds according to claim 1, selected from the group consisting of: N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N '- cyclohexylurea; A / - (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) - / \ / - (1-adamantyl) urea; A / - (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) - / \ / - (tetrahydro-2-pyranyl) urea; / N- [4-chloro-2-hydroxy-3-sulfamylphenyl] - / \ / - (3-tetrahydrofuryl) urea; 6-chloro-2-hydroxy-3- [3- (2-methyl-cyclopropyl) -ureido] -benzenesulfonamide; A / - [4-chloro-2-hydroxy-3-sulfamylphenyl] - / V-cyclohexylurea; 6-chloro-2-hydroxy-3- [3- (2,2,3,3-tetramethyl-cyclopropyl) -ureido] -benzenesulfonamide; 6-chloro-2-hydroxy-3- (3-piperidin-4-yl-ureido) -benzenesulfonamide; And / - [4-chloro-2-Hydroxy-3-sulfamylphenyl] -N / - (4-methyl-cyclohexyl) urea; 6-chloro-2-hydroxy-3- [3- (3-methoxy-cyclohexyl) -ureido] -benzenesulfonamide; A / - [4-chloro-2-hydroxy-3-sulfamylphenyl] - / \ / '- cyklopentylmočovinu; / N- [4-chloro-2-hydroxy-3-sulfamylphenyl] - / V-cyklobutylmočovinu; A / - [4-chloro-2-hydroxy-3-sulfamylphenyl] - / Y-cyclopropyl-urea; 4- [6-Chloro-3- (3-cyclopentylureido) -2-hydroxybenzenesulfonyl] piperazine-1-carboxylic acid tert-butyl ester; 1- [4-chloro-2-hydroxy-3- (piperazine-1-sulfonyl) -phenyl] -3-cyclopentyl-urea; 4- [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-A /, A / '- dimethyl-benzenesulfonamide. The sulfonamide-substituted diphenylurea compound of claim 7 which is N- (3-aminosulfonyl-4-chloro-2-hydroxyphenyl) -N-cyclohexylurea. The sulfonamide substituted diphenylurea compound of claim 7, which is the compound in the form of its sodium salt. The sulfonamide substituted diphenylurea compound according to claim 7, which is the compound in the form of its potassium salt. -41 A pharmaceutical composition comprising a sulfonamide substituted diphenylurea compound according to any one of claims 1 to 10 and a pharmaceutically acceptable carrier or diluent. Use of the sulfonamide substituted diphenylurea compounds according to any one of claims 1 to 10 for the manufacture of a medicament for the treatment of a chemokine-mediated disease, wherein the chemokine binds to an IL-8 and / or β receptor of a mammal. The use of claim 12, wherein the chemokine mediated disease is selected from the group consisting of: psoriasis, atopic dermatitis, osteoarthritis, rheumatoid arthrosis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, ulcerative colitis, ulcerative colitis, , septic shock, multiple sclerosis, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, host versus graft reaction, Alzheimer's disease, rejection of allograft, angina, restorative, malaria, restorative, malaria unwanted hematopoietic stem cell release and diseases caused by respiratory tract viruses, herpes viruses and hepatitis viruses, meningitis, cystic fibrosis, premature labor, cough, itching, multi-organ dysfunction, trauma, tensions, stomach those bruises, psoriatic arthritis, herpes, encephalitis, CNS vasculitis, traumatic brain damage, CNS tumors, subarachnoid haemorrhage, post-operative trauma, interstitial lung inflammation, hypersensitivity, crystal-induced enteritis, chronic acute arthritis, acute arthritis, acute arthritis, sinusitis, uveitis, polymyositis, vasculitis, acne, gastric and duodenal ulcers, celiac disease, esophagitis, glositis, airway obstruction, respiratory hyperreaction, bronchiolitis, pulmonary inflammation, bronchiolusitis, bronchiochiitis, bronchiochiitis, bronchiochiitis, bronchiochiitis, bronchiochiitis hypercapnia, hyperinflation, hypoxemia, hyperoxia-induced inflammation, hypoxia, lung surgery, pulmonary -42 fibrosis, pulmonary hypertension, ventricular hypertropy, sarcoidosis, small airway disease, ventilation-perfusion disorders, wheezing, rhinitis and lupus.