WO2000072800A2 - Il-8 receptor antagonists - Google Patents

Il-8 receptor antagonists Download PDF

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Publication number
WO2000072800A2
WO2000072800A2 PCT/US2000/014655 US0014655W WO0072800A2 WO 2000072800 A2 WO2000072800 A2 WO 2000072800A2 US 0014655 W US0014655 W US 0014655W WO 0072800 A2 WO0072800 A2 WO 0072800A2
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alkyl
optionally substituted
alkenyl
aryl
heteroaryl
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PCT/US2000/014655
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French (fr)
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WO2000072800A3 (en
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Michael R. Palovich
Katherine L. Widdowson
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Smithkline Beecham Corporation
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/20—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carbonic acid, or sulfur or nitrogen analogues thereof
    • C07D295/215—Radicals derived from nitrogen analogues of carbonic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C279/00—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
    • C07C279/18—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to carbon atoms of six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C281/00—Derivatives of carbonic acid containing functional groups covered by groups C07C269/00 - C07C279/00 in which at least one nitrogen atom of these functional groups is further bound to another nitrogen atom not being part of a nitro or nitroso group
    • C07C281/16—Compounds containing any of the groups, e.g. aminoguanidine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/20—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by singly bound oxygen or sulphur atoms
    • C07D211/22—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by singly bound oxygen or sulphur atoms by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/26—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by nitrogen atoms

Definitions

  • This invention relates to a novel group of tetraalkyl guanidine compounds, processes for the preparation thereof, the use thereof in treating IL-8. GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2. and ENA-78 mediated diseases and pharmaceutical compositions for use in such therapy. BACKGROUND OF THE INVENTION
  • Interleukin-8 Interleukin-8
  • NAP-1 neutrophil attractant/activation protein- 1
  • MDNCF monocyte derived neutrophil chemotactic factor
  • NAF neutrophil activating factor
  • T-cell lymphocyte chemotactic factor T-cell lymphocyte chemotactic factor.
  • Interleukin-8 is a chemoattractant for neutrophils, basophils, and a subset of T-cells. It is produced by a majority of nucleated cells including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-loc, IL-l ⁇ or LPS, and by neutrophils themselves when exposed to LPS or chemotactic factors such as FMLP.
  • GRO ⁇ , GRO ⁇ and NAP-2 also belong to the chemokine ⁇ family. Like IL-8 these chemokines have also been referred to by different names. For instance GRO ⁇ , ⁇ , ⁇ have been referred to as MGSA ⁇ , ⁇ and ⁇ respectively (Melanoma Growth Stimulating Activity), see Richmond et al. J. Cell Physiology
  • IL-8, Gro ⁇ . GRO ⁇ , GRO ⁇ , NAP-2 and ENA-78 stimulate a number of functions in vitro. They have all been shown to have chemoattractant properties for neutrophils, while IL-8 and GRO ⁇ have demonstrated T-lymphocytes, and basophiles chemotactic activity. In addition IL-8 can induce histamine release from basophils from both normal and atopic individuals. GRO- ⁇ and IL-8 can in addition, induce lysozomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mac- 1 (CD l ib/CD 18) on neutrophils without de novo protein synthesis.
  • ELR chemokines (those containing the amino acids ELR motif just prior to the CXC motif) have also been implicated in angiostasis, Strieter et al, Science 258, 1798 (1992).
  • IL-8, Gro ⁇ , GRO ⁇ , GRO ⁇ , and NAP-2 induce neutrophil shape change, chemotaxis, granule release, and respiratory burst, by binding to and activating receptors of the seven-transmembrane, G-protein-linked family, in particular by binding to IL-8 receptors, most notably the B-receptor, 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 a review see R. Freidinger in: Progress in Drug Research. Vol. 40, pp. 33-98. Birkhauser Verlag, Basel 1993.
  • the IL-8 receptor represents a promising target for the development of novel anti-inflammatory agents.
  • IL-8R ⁇ which binds only IL-8 with high affinity
  • IL-8RB which has high affinity for IL-8 as well as for GRO- ⁇ , GRO ⁇ , GRO ⁇ and NAP-2.
  • IL-8R ⁇ which binds only IL-8 with high affinity
  • IL-8RB which has high affinity for IL-8 as well as for GRO- ⁇ , GRO ⁇ , GRO ⁇ and NAP-2.
  • This invention provides for a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an EL-8 ⁇ or ⁇ receptor and which method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the chemokine is IL-8.
  • This invention also relates to a method of inhibiting the binding of IL-8 to its receptors in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula (I).
  • R is OH, SH, NHSO 2 Rd
  • Ri is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; C i-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CRgRg)q S(O) t R4; hydroxy; hydroxy C i-4alkyl; aryl; aryl Ci-4 alkyl; aryloxy.
  • R 8 is hydrogen or Ci-4 alkyl; Rio is C MO alkyl C(O)2R 8 ;
  • Rl 1 is hydrogen, C1-.4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylCi-4alkyl, optionally substituted heterocychc, or optionally substituted heterocyclicCi-4alkyl;
  • R12 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl;
  • Rl3 and R 14 are independently hydrogen, optionally substituted Ci-4 alkyl or one of R13 and R14 may be optionally substituted aryl;
  • Rl7 is Ci_4alkyl. aryl, arylalkyl, heteroaryl, heteroarylC ⁇ _4alkyl, heterocychc, or heterocyclicC ⁇ _4alkyl, wherein the aryl, heteroaryl and heterocychc rings may all be optionally substituted;
  • Rd is NRgR , alkyl, arylCl-4alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-C ⁇ _4alkyl, heteroarylC2-4 alkenyl, heterocychc, heterocyclicC ⁇ .4 alkyl, wherein the aryl, heteoaryl and heterocychc rings may all be optionally substituted.
  • the compounds of Formula (I) may also be used in association with the veterinary treatment of mammals, other than humans, in need of inhibition of IL-8 or other chemokines which bind to the IL-8RA and RB receptors.
  • Chemokine mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section.
  • Ri is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Cj-io alkyl, such as CF3; Ci-io alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C2-10 alkenyl; Ci-io alkoxy, such as methoxy, or ethoxy; halosubstituted Ci-io alkoxy, such as trifluoromethoxy; azide; (CR Rg)q S(O) t R4, wherein t is 0, 1 or 2; hydroxy; hydroxy Ci-4alkyl, such as methanol or ethanol; aryl, such as phenyl or naphthyl; aryl Ci-4 alkyl, such as benzyl; aryloxy.
  • aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl. heterocychc, heterocyclicalkyl, and heterocyclicalkenyl moieties may all be optionally substituted as defined herein below.
  • q is 0, or an integer having a value of 1 to 10.
  • Ri forms a dioxybridge
  • s is preferably 1.
  • Rl forms an additional unsaturated ring, it is preferably 6 membered resulting in a naphthylene ring system. This naphthylene ring may be substituted independently, 1 to 3 times by the other R moieties as defined above.
  • R4 and R5 are independently hydrogen, optionally substituted C ⁇ _4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C i -4-dkyl, heterocychc, heterocyclicCi-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from O/N/S.
  • R 8 is suitably independently selected from hydrogen or C 1.4 alkyl.
  • RlO is suitably -io alkyl C(O)2R 8 , such as CH2C(O)2H or
  • Rl 1 is suitably hydrogen, C1.4 alkyl, aryl, aryl C ⁇ _4 alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocychc, or heterocychc C ⁇ _4alkyl.
  • R12 is suitably hydrogen, Cl-10 alkyl, optionally substituted aryl or optionally substituted arylalkyl.
  • Rl7 is suitably Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylCi-4alkyl, heterocychc, or heterocyclicCi-4alkyl, wherein the aryl, heteroaryl and heterocychc rings may all be optionally substituted.
  • Ri is halogen, cyano. nitro, CF3, C(O)NR4R5, alkenyl
  • R13 and R 14 are independently hydrogen, optionally substituted C ⁇ _4 alkyl which may be straight or branched as defined herein, or one of R13 and R 14 are an optionally substituted aryl; v is an integer having a value of 1 to 4.
  • R13 or R 4 are an optionally substituted alkyl
  • the alkyl moiety may be substituted one to three times independently by halogen; halosubstituted Ci_4 alkyl such as trifluromethyl; hydroxy; hydroxy C ⁇ _4alkyl, Ci-4 alkoxy; such as methoxy, or ethoxy.
  • halosubstituted Ci-io alkoxy S(O)tR4; aryl; NR4R5; NHC(O)R4; C(O)NR4R5; or C(O)OR 8 .
  • Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR 8 R 8 )q S(O)tR4; hydroxy; hydroxyCi-4alkyl: aryl; aryl Ci-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocychc, heterocychc C ⁇ _4alkyl; aryl C2-10 alkenyl; heteroaryl C2-IO alkenyl; heterocychc C2-10 alkenyl; (CR 8 Rg)q NR4R5; C2-10 alkenyl C(O)NR4R5; (CR 8 R 8 )q C(O)NR4R
  • Y When Y forms an additional unsaturated ring, it is preferably 6 membered resulting in a naphthylene ring system.
  • This naphthylene ring may be substituted 1 to 3 times by other Y moieties as defined above.
  • the aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl. heterocychc, heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.
  • Rd is a NR5R7, alkyl, aryl C1.4 alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-C 1.4 alkyl, heteroarylC2-4 alkenyl, heterocychc, heterocyclicC ⁇ .4 alkyl, or heterocychc C 2-4 alkenyl moiety, wherein the aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heterocychc, and heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.
  • Y is preferably a halogen, Ci-4 alkoxy, optionally substituted aryl, optionally substituted aryloxy or arylalkoxy, methylene dioxy, NR4R5, thio C ⁇ _ 4alkyl, thioaryl, halosubstituted alkoxy, optionally substituted C1-.4 alkyl, or hydroxy alkyl.
  • Y is more preferably mono-substituted halogen, disubstituted halogen, mono-substituted alkoxy, disubstituted alkoxy. methylenedioxy, aryl, or alkyl, more preferably these groups are mono or di-substituted in the 2'- position or
  • Y may be substituted in any of the 5 ring positions, preferably when R is OH, SH, or NHSO2Rd» is preferably mono-substituted in the 2'-position or 3 - position, with the 4'- preferably being unsubstituted. If the ring is disubstituted, when R is OH, SH, or NSO2R , substituents are preferably in the 2' or 3' position of a monocyclic ring. While both Ri and Y can both be hydrogen, it is preferred that at least one of the rings be substituted, and more preferably that both rings are substituted.
  • Exemplified compounds of Formula (I) include:
  • "optionally substituted” unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted Ci-ioalkyl; Ci-io alkoxy, such as methoxy or ethoxy; S(O)m' Ci-io alkyl, wherein m' is 0, 1 or 2, such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono & di-substituted amino, such as in the NR4R5 group; NHC(O)R4; C(O)NR4R5; C(O)OH; S(O)2NR4Rs; NHS(O)2Rl5, Ci-io alkyl, such
  • Another aspect of the present invention are the novel compounds of Formula (II), or a pharmaceutically acceptable salt thereof, as described below, which are also useful in inhibiting the binding of IL-8 to its receptors in a mammal in need thereof.
  • This invention also relates to the pharmaceutical compositions comprising a compound of Formula (II) and a pharmaceutically acceptable diluent or carrier.
  • Compounds of Formula (II) are also useful for treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8RA or RB receptor and which method comprises administering an effective amount of a compound of Formula (II) or a pharmaceutically acceptable salt thereof.
  • Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of inorganic and organic acids, such as hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane sulphonic acid, ethane sulphonic 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 mandelic acid.
  • pharmaceutically acceptable salts of compounds of Formula (I) may also be formed with a pharmaceutically acceptable cation, for instance, if a substituent group comprises a carboxy moiety.
  • Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations.
  • halo all halogens, that is chloro, fluoro, bromo and iodo.
  • C ⁇ _ ⁇ oalkyl or “alkyl” both straight and branched chain alkyls of 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, iso-butyl, tert-butyl, n- pentyl and the like.
  • cycloalkyl is used herein to mean cyclic alkyls, preferably of 3 to 8 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl, and the like.
  • alkenyl is used herein at all occurrences to mean straight or branched chain alkyls of 2-10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l- propenyl, 1-butenyl, 2-butenyl and the like.
  • heteroaryl (on its own or in any combination, such as “heteroaryloxy”, or “heteroaryl alkyl”) - 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, thiazole, thiadiazole, triazole, imidazole, or benzimidazole.
  • heterocyclicalkyl 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; such as, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, or imidazolidine.
  • arylalkyl or “heteroarylalkyl” or “heterocyclicalkyl” is used herein to mean Ci-io alkyl, as defined above, attached to an aryl. heteroaryl or heterocychc moiety, as also defined herein, unless otherwise indicated.
  • Rl moieties or two Y moieties may together form a 5 or 6 membered unsaturated ring
  • a napthylene ring system or a phenyl moiety having attached a 6 membered partially unsaturated ring such as a C cycloalkenyl, i.e hexene, or a C5 cyloalkenyl moiety, cyclopentene.
  • the compounds of Formula (I) may be obtained by applying synthetic procedures, some of which are illustrated in the Schemes below.
  • the synthesis provided for in these Schemes is applicable for the producing compounds of Formula (I) having a variety of different R, Rl, and aryl groups which are reacted, employing optional substituents, which are suitably protected, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, in those cases, then affords compounds of the nature generally disclosed.
  • further compounds of these formulas may be prepared by applying standard techniques for functional group interconversion, well known in the art. While the schemes are shown with compounds only of Formula (I) this is merely for illustration purposes only.
  • the desired aniline 6-scheme- 1 can be prepared from the commercially available benzoxazolinone 1 -scheme- 1.
  • Bromide 2-scheme- 1 can be prepared from benzoxazolinone 1 -scheme- 1 using standard bromination conditions such as bromine and sodium acetate in acetic acid.
  • Bromide 2-scheme- 1 can be converted to the cyanide 3-scheme-l using standard procedures such as copper (I) cyanide in refluxing DMF.
  • the amide 3-scheme-l can be converted to the BOC protected compound 4-scheme-l using standard conditions such as BOC anhydride and triethylamine with a catalytic amount of dimethylaminopyridine in methylene chloride or another suitable organic solvent.
  • the oxazolinone 4-scheme- 1 can be converted to the desired aniline 6-scheme- 1 by first hydrolysis to the phenol 5; scheme- 1 using standard conditions such as potassium carbonate in methanol followed by removal of the BOC protecting group using standard conditions such as trifluoroacetic acid in methylene chloride or another suitable organic solvent to give the aniline 6-scheme- 1.
  • Scheme 2
  • the desired thiourea 3-scheme-2 can be prepared as outlined in scheme 2.
  • the aniline 1 -scheme-2 can be coupled with a commercially available isothiocyanate or with an isothiocyanate made from condensing a commercially available amine with thiophosgene or a thiophosgene equivalent to give the thiourea 2-scheme-2.
  • the phenol 2-scheme-2 can be protected as the TBS ether 3-scheme-2 using standard conditions such as TBSCl and imidazole in THF or another suitabel organic solvent.
  • the amine 1 -scheme-2 can be converted to the desired isothiocyanate by first protecting the phenol with a suitable protecting group such as the TBS ether 4-scheme-2 using standard conditions such as TBSCl and an amine base such as imidazole in THF or another suitable organic solvent.
  • the thiourea 5 scheme-2 can be made by condensing the amine 4-scheme 2 with thiophosgene in the presence of base such as potassium carbonate.
  • the desired thiourea 3-scheme-2 can be prepared from the isothiocyanate 5-scheme-2 by condensing it with the desired amine in a suitable organic solvent such as ethanol or DMF.
  • the desired tetraalkylguanidine 4-scheme-3 can be prepared as outlined in scheme 3.
  • the carbodiimide 2-scheme-3 can be prepared from the thiourea L scheme-3 under suitable conditions such as using an excess amount of methane sulfonyl chloride and a suitable amine base such as triethylamine in an organic solvent preferably methylene chloride at room temperature.
  • the guanidine compound 3-scheme-3 can be obtained by utilizing standard procedures such as condensing the carbodiimide 2-scheme-3 and the desired secondary amine in a suitable solvent such as acetonitrile or THF at room temperature.
  • the desired guanidine 4-scheme-3 can be obtained from 3-scheme-3 by removal of the TBS protecting group using standard conditions such as TBAF in THF under low reaction temperatures.
  • the compounds of Formula (I) and (II), or a pharmaceutically acceptable salt thereof can be used in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or other mammal, which is exacerbated or caused by excessive or unregulated IL-8 cytokine production by such mammal's cell, such as but not limited to monocytes and or macrophages, or other chemokines which bind to the IL-8 ⁇ or ⁇ receptor, also referred to as the type I or type II receptor.
  • the present invention provides a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8 ⁇ or ⁇ receptor and which method comprises administering an effective amount of a compound of Formula (I) or (II) or a pharmaceutically acceptable salt thereof.
  • the chemokines are IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78.
  • the compounds of Formula (I) are administered in an amount sufficient to inhibit cytokine function, in particular IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78, such that they are biologically regulated down to normal levels of physiological function, or in some case to subnormal levels, so as to ameliorate the disease state.
  • Abnormal levels of IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 for instance in the context of the present invention constitute: (I) levels of free IL-8 greater than or equal to 1 picogram per mL; (ii) any cell associated IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 above normal physiological levels; or (iii) the presence EL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 above basal levels in cells or tissues in IL-8, GRO ⁇ . GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 respectively, is produced.
  • Chemokine mediated diseases include psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, stroke, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft vs. host reaction, alzheimers disease, allograft rejections, malaria, restinosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis or undesired hematopoietic stem cells release.
  • IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ or NAP-2 has the unique property of promoting neutrophil chemotaxis, enzyme release including but not limited to elastase release as well as superoxide production and activation.
  • the ⁇ -chemokines but particularly, GRO ⁇ , GRO ⁇ , GRO ⁇ or NAP-2, working through the IL-8 type I or II receptor can promote the neovascularization of tumors by promoting the directional growth of endothelial cells. Therefore, the inhibition of IL-8 induced chemotaxis or activation would lead to a direct reduction in the neutrophil infiltration. Recent evidence also implicates the role of chemokines in the treatment of HIV infections, Littleman et al., Nature 381, pp 661 (1996) and Koup et al., Nature 381, pp 667 (1996).
  • the present invention also provides for a means of treating, in an acute setting, as well as preventing, in those individuals deemed susceptible to, CNS injuries by the chemokine receptor antagonist compounds of Formula (I).
  • CNS injuries as defined herein include both open or penetrating head trauma, such as by surgery, or a closed head trauma injury, such as by an injury to the head region. Also included within this definition is ischemic stroke, particularly to the brain area.
  • Ischemic stroke may be defined as a focal neurologic disorder that results from insufficient blood supply to a particular brain area, usually as a consequence of an embolus. thrombi, or local atheromatous closure of the blood vessel.
  • embolus. thrombi, or local atheromatous closure of the blood vessel The role of inflammatory cytokines in this area has been emerging and the present invention provides a mean for the potential treatment of these injuries. Relatively little treatment, for an acute injury such as these has been available.
  • TNF- ⁇ is a cytokine with proinflammatory actions, including endothelial leukocyte adhesion molecule expression.
  • Leukocytes infiltrate into ischemic brain lesions and hence compounds which inhibit or decrease levels of TNF would be useful for treatment of ischemic brain injury. See Liu et al., Stroke, Vol. 25., No. 7, pp 1481-88 (1994) whose disclosure is incorporated herein by reference.
  • the compounds of Formula (I) are administered in an amount sufficient to inhibit binding to the IL-8 alpha or beta receptors such as evidenced by a reduction in neutrophil chemotaxis and activation.
  • the discovery that the compounds of Formula (I) are inhibitors of IL-8 binding is based upon the effects of the compounds of Formulas (I) in the in vitro receptor binding assays which are described herein.
  • the compounds of Formula (I) have been shown, in some instances, to be dual inhibitors of both recombinant type I and type II IL-8 receptors.
  • the compounds are inhibitors of only one receptor, more preferably Type II.
  • IL-8 mediated disease or disease state refers to any and all disease states in which IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 plays a role, either by production of IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 themselves, or by IL-8, GRO ⁇ , GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF.
  • a disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by IL-8.
  • chemokine mediated disease or disease state refers to any and all disease states in which a chemokine which binds to an IL-8 ⁇ or ⁇ receptor plays a role, such as but not limited IL-8, GRO ⁇ . GRO ⁇ , GRO ⁇ , NAP-2 or ENA-78. This would include a disease state in which, IL-8 plays a role, either by production of IL-8 itself, or by IL-8 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF.
  • cytokine refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response.
  • a cytokine includes, but is not limited to, monokines and lymphokines, regardless of which cells produce them.
  • a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte.
  • Lymphokines are generally referred to as being produced by lymphocyte cells.
  • cytokines include, but are not limited to, Interleukin- 1 (IL-1), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Tumor Necrosis Factor-alpha (TNF- ⁇ ) and Tumor Necrosis Factor beta (TNF- ⁇ ).
  • chemokine refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response, similar to the term “cytokine” above.
  • a 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.
  • chemokines include, but are not limited to. IL-8, GRO- ⁇ , GRO- ⁇ , GRO- ⁇ , NAP-2, ENA-78, IP- 10, MlP-l ⁇ , MlP- ⁇ , PF4, and MCP 1, 2, and 3.
  • a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy it will normally be formulated into a pharmaceutical , composition in accordance with standard pharmaceutical practice.
  • This 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.
  • Compounds of Formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation.
  • the compounds of Formula (I) may be administered in conventional dosage forms prepared by combining a compound of Formula (I) with standard pharmaceutical carriers according to conventional procedures.
  • the compounds of Formula (I) may also be administered in conventional dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation.
  • the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables.
  • the carrier(s) 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 employed may be, for example, either a solid or liquid.
  • solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like.
  • liquid carriers are syrup, peanut oil, olive oil, water and the like.
  • the carrier or diluent may include time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax.
  • time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax.
  • a wide variety of pharmaceutical forms can be employed.
  • the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge.
  • the amount of solid carrier will vary widely but preferably will be from about 25mg. to about lg.
  • the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.
  • Compounds of Formula (I) may be administered topically, that is by non- systemic administration. This includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream.
  • systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.
  • Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose.
  • the active ingredient may comprise, for topical administration, from 0.001% to 10% w/w, for instance from 1% to 2% by weight of the Formulation. It may however comprise as much as 10% w/w but preferably will comprise less than 5% w/w, more preferably from 0.1% to 1% w/w of the Formulation.
  • Lotions according to the present invention include those suitable for application to the skin or eye.
  • An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those for the preparation of drops.
  • Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.
  • Creams, ointments or pastes according to the present invention are semi-solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with the aid of suitable machinery, with a greasy or non-greasy base.
  • the base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as steric or oleic acid together with an alcohol such as propylene glycol or a macrogel.
  • the formulation may incorporate any suitable surface active agent such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof.
  • Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.
  • Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and preferably including a surface active agent.
  • the resulting solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98-100 °C. for half an hour.
  • the solution may be sterilized by filtration and transferred to the container by an aseptic technique.
  • bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%).
  • Suitable solvents for the preparation of an oily solution include glycerol. diluted alcohol and propylene glycol.
  • Compounds of formula (I) may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration.
  • the subcutaneous and intramuscular forms of parenteral administration are generally preferred.
  • Appropriate dosage forms for such administration may be prepared by conventional techniques.
  • Compounds of Formula (I) may also be administered by inhalation, that is by intranasal and oral inhalation administration.
  • Appropriate dosage forms for such administration such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.
  • the daily oral dosage regimen will preferably be from about 0.01 to about 80 mg/kg of total body weight.
  • the daily parenteral dosage regimen about 0.001 to about 80 mg/kg of total body weight.
  • the daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times daily.
  • the daily inhalation dosage regimen will preferably be from about 0.01 mg/kg to about 1 mg/kg per day.
  • the optimal quantity and spacing of individual dosages of a compound of Formula (I) or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.
  • IL-8, and Gro- ⁇ chemokine inhibitiory effects of compounds of the present invention were determined by the following in vitro assay: Receptor Binding Assays: [125l] IL-8 (human recombinant) was obtained from Amersham Corp.,
  • homogenization buffer was changed to lOmM Tris-HCL, lmM MgS04, 0.5mM EDTA (ethylene-diaminetetra- acetic acid), ImMPMSF ( ⁇ -toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH 7.5.
  • Membrane protein concentration was determined using Pierce Co. micro-assay kit using bovine serum albumin as a standard. All assays were performed in a 96-well micro plate format.
  • Each reaction mixture contained 125 ⁇ iL_g (0.25 nM) or 125 ⁇ Gro- ⁇ and 0.5 ⁇ g/mL of IL-8R ⁇ or 1.0 ⁇ g/mL of IL-8R ⁇ membranes in 20 mM Bis- Trispropane and 0.4 mM Tris HCl buffers, pH 8.0, containing 1.2 mM MgSO4, 0.1 mM EDTA, 25 mM NaCl and 0.03% CHAPS.
  • drug or compound of interest was added which had been pre-dissolved in DMSO so as to reach a final concentration of between 0.0 InM and 100 uM.
  • the assay was initiated by addition of 12-5I-IL-8. After 1 hour at room temperature the plate was harvested using a Tomtec 96-well harvester onto a glass fiber filtermat blocked with 1 % polyethylenimine/0.5% BSA and washed 3 times with 25 mM NaCl, 10 mM
  • the recombinant IL-8 R ⁇ , or Type I, receptor is also referred to herein as the non- permissive receptor and the recombinant IL-8 R ⁇ , or Type II, receptor is referred to as the permissive receptor.
  • the in vitro inhibitory properties of these compounds are determined in the neutrophil chemotaxis assay as described in Current Protocols in Immunology, vol I, Suppl 1, Unit 6.12.3., whose disclosure is incorporated herein by reference in its entirety.
  • Neutrophils where isolated from human blood as described in Current Protocols in Immunology Vol I, Suppl 1 Unit 7.23.1, whose disclosure is incorporated herein by reference in its entirety.
  • the chemoattractants IL-8, GRO- ⁇ , GRO- ⁇ , GRO- ⁇ and NAP-2 are placed in the bottom chamber of a 48 multiwell chamber (Neuro Probe, Cabin John, MD) at a concentration between 0.1 and 100 nM. The two chambers are separated by a 5um polycarbonate filter.
  • the compounds of this invention are tested for their ability to prevent Elastase release from human neutrophils.
  • 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 (NaCl 118, KC14.56, NaHCO3 25, KH2PO4 1.03, Glucose 11.1, HEPES 5 mM, pH 7.4) are placed in each well of a 96 well plate in a volume of 50 ul.
  • the test compound (0.001 - 1000 nM) in a volume of 50 ul
  • Cytochalasin B in a volume of 50 ul (20ug/ml)
  • Ringers buffer in a volume of 50 ul.
  • the present assay provides for examination of the expression of tumor necrosis factor mRNA in specfic brain regions which follow experimentally induced lateral fluid-percussion traumatic brain injury (TBI) in rats.
  • TBI experimentally induced lateral fluid-percussion traumatic brain injury
  • LC left (injured) parietal cortex
  • RC contralateral right cortex
  • LA cortex adjacent to injured parietal cortex
  • RA right cortex
  • RH right hippocampus
  • TNF- ⁇ mRNA expression is observed in LH (104 ⁇ 17% of positive control, p ⁇ 0.05 compared with sham), LC (105 ⁇ 21%, p ⁇ 0.05) and LA (69 ⁇ 8%, p ⁇ 0.01) in the traumatized hemisphere 1 hr. following injury.
  • An increased TNF- ⁇ mRNA expression is also observed in LH (46 ⁇ 8%, p ⁇ 0.05), LC (30 ⁇ 3%, p ⁇ 0.01) and LA (32 ⁇ 3%, p ⁇ 0.01) at 6 hr. which resolves by 24 hr. following injury.
  • TNF- ⁇ mRNA In the contralateral hemisphere, expression of TNF- ⁇ mRNA is increased in RH (46 ⁇ 2%, p ⁇ 0.01), RC (4 ⁇ 3%) and RA (22 ⁇ 8%) at 1 hr. and in RH (28 ⁇ 11%), RC (7 ⁇ 5%) and RA (26 ⁇ 6%, p ⁇ 0.05) at 6 hr. but not at 24 hr. following injury. In sham (surgery without injury) or naive animals, no consistent changes in expression of TNF- ⁇ mRNA are observed in any of the 6 brain areas in either hemisphere at any times.
  • TNF- ⁇ mRNA is altered in specific brain regions, including those of the non-traumatized hemisphere. Since TNF- ⁇ is able to induce nerve growth factor (NGF) and stimulate the release of other cytokines from activated astrocytes, this post-traumatic alteration in gene expression of TNF- ⁇ plays an important role in both the acute and regenerative response to CNS trauma.
  • NGF nerve growth factor
  • This assay characterizes the regional expression of interleukin-l ⁇ (IL-l ⁇ ) mRNA in specific brain regions following experimental lateral fluid-percussion traumatic brain injury (TBI) in rats.
  • TBI lateral fluid-percussion traumatic brain injury
  • LC left (injured) parietal cortex
  • RC contralateral right cortex
  • LA cortex adjacent to injured parietal cortex
  • RA right cortex
  • LH left hippocampus
  • RH right hippocampus

Abstract

This invention relates to the novel use of phenyl ureas in the treatment of disease states mediated by the chemokine, Interleukin-8 (IL-8).

Description

IL-8 RECEPTOR ANTAGONISTS FIELD OF THE INVENTION
This invention relates to a novel group of tetraalkyl guanidine compounds, processes for the preparation thereof, the use thereof in treating IL-8. GROα, GROβ, GROγ, NAP-2. and ENA-78 mediated diseases and pharmaceutical compositions for use in such therapy. BACKGROUND OF THE INVENTION
Many different names have been applied to Interleukin-8 (IL-8), such as neutrophil attractant/activation protein- 1 (NAP-1), monocyte derived neutrophil chemotactic factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte chemotactic factor. Interleukin-8 is a chemoattractant for neutrophils, basophils, and a subset of T-cells. It is produced by a majority of nucleated cells including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-loc, IL-lβ or LPS, and by neutrophils 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).
Gro , GROβ, GROγ and NAP-2 also belong to the chemokine α family. Like IL-8 these chemokines have also been referred to by different names. For instance GROα, β, γ have been referred to as MGSAα, 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 of the chemokines of the α-family which possess the ELR motif directly preceding the CXC motif bind to the IL-8 B receptor.
IL-8, Groα. GROβ, GROγ, NAP-2 and ENA-78 stimulate a number of functions in vitro. They have all been shown to have chemoattractant properties for neutrophils, while IL-8 and GROα have demonstrated T-lymphocytes, and basophiles chemotactic activity. In addition IL-8 can induce histamine release from basophils from both normal and atopic individuals. GRO-α and IL-8 can in addition, induce lysozomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mac- 1 (CD l ib/CD 18) on neutrophils without de novo protein synthesis. This may contribute to increased adhesion of the neutrophils to vascular endothelial cells. Many known diseases are characterized by massive neutrophil infiltration. As IL-8, Groα, GROβ, GROγ and NAP-2 promote the accumulation and activation of neutrophils, these chemokines have been implicated 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. Pis. 146. 427 (1992); Donnely et al.. Lancet 341. 643 (1993). In addition the ELR chemokines (those containing the amino acids ELR motif just prior to the CXC motif) have also been implicated in angiostasis, Strieter et al, Science 258, 1798 (1992).
In vitro, IL-8, Groα, GROβ, GROγ, and NAP-2 induce neutrophil shape change, chemotaxis, granule release, and respiratory burst, by binding to and activating receptors of the seven-transmembrane, G-protein-linked family, in particular by binding to IL-8 receptors, most notably the B-receptor, 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 a review see R. Freidinger in: Progress in Drug Research. Vol. 40, pp. 33-98. Birkhauser Verlag, Basel 1993. Hence, the IL-8 receptor represents a promising target for the development of novel anti-inflammatory agents.
Two high affinity human IL-8 receptors (77% homology) have been characterized: IL-8Rα, which binds only IL-8 with high affinity, and IL-8RB, which has high affinity for IL-8 as well as for GRO-α, GROβ, GROγ 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, in this field, for compounds which are capable of binding to the IL-8 α or β receptor. Therefore, conditions associated with an increase in IL-8 production (which is responsible for chemotaxis of neutrophil and T-cells subsets into the inflammatory site) would benefit by compounds which are inhibitors of IL-8 receptor binding.
SUMMARY OF THE INVENTION This invention provides for a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an EL-8 α or β receptor and which method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In particular the chemokine is IL-8.
This invention also relates to a method of inhibiting the binding of IL-8 to its receptors in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula (I).
Compounds of Formula (I) useful in the present invention are represented by the structure:
Figure imgf000004_0001
(I) wherein:
R is OH, SH, NHSO2Rd
Ri is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; C i-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CRgRg)q S(O)tR4; hydroxy; hydroxy C i-4alkyl; aryl; aryl Ci-4 alkyl; aryloxy. aryl C i -4 alkyloxy; heteroaryl; heteroarylalkyl; heterocychc, heterocychc C [-4alkyl; heteroaryl C ι_4 alkyloxy; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl: heterocychc C2- 10 alkenyl; (CRgRg)qNR4R5; C2-10 alkenyl C(O)NR4R5: (CR8R8)q C(O)NR4R5; (CR8Rg)q C(O)NR4Rl0; S(O)3H; S(O)3R8; (CR8R8)q C(O)Rι 1 ; C2- IO alkenyl C(O)R ι 1 ; C2-10 alkenyl
C(O)ORι l(CR8R8)q C(O)ORι2; (CR8R8)q OC(O) Ri 1 ; (CRgR8)qNR4C(O)Ri 1 , (CR8R8)q NHS(O)2Ri7, (CRgRg)q S(O)2NR4R5; or two Rj moieties together may form O-(CH2)s - or a 5 to 6 membered unsaturated ring; q is 0, or an integer having a value of 1 to 10; t is 0, or an integer having a value of 1 or 2; s is an integer having a value of 1 to 3; v is an integer having a value of 0 to 4; R4 and R5 are independently hydrogen, optionally substituted C i-4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C1.4a.kyl, heterocychc, heterocyclicC 1 -4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur; R6 and R7 are independently hydrogen or a C i-4 alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur; Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci -io alkyl: C i-io alkyl; C2-10 alkenyl; -io alkoxy; halosubstituted Ci -io alkoxy; azide: (CR8R8)q S(O)tR4; hydroxy; hydroxyCi-4alkyl; aryl; aryl C1 -4 alkyl; aryloxy; arylCι.4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocychc, heterocychc Ci-4alkyl; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocychc C2-10 alkenyl; (CR8Rg)q NR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)q C(O)NR4R5; (CRgRg)q C(O)NR4Rl0; S(O)3H; S(O)3R8; (CR8R8)q C(O)Rι 1; C2-10 alkenyl C(O)Rι 1 ; C2-10 alkenyl C(O)OR 1 1 ; C(O)R 1 1 ; (CR8R8)q C(O)OR 12; (CR8R8)q OC(O) R 11 ; (CR8R8)q NR4C(O)Rι 1 , (CR8Rg)q NHS(O)2Rd, (CR8R8)q S(O)2NR4R5; or two Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; n is an integer having a value of 1 to 3; m is an integer having a value of 1 to 3;
R8 is hydrogen or Ci-4 alkyl; Rio is C MO alkyl C(O)2R8;
Rl 1 is hydrogen, C1-.4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylCi-4alkyl, optionally substituted heterocychc, or optionally substituted heterocyclicCi-4alkyl; R12 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl3 and R 14 are independently hydrogen, optionally substituted Ci-4 alkyl or one of R13 and R14 may be optionally substituted aryl;
Rl7 is Ci_4alkyl. aryl, arylalkyl, heteroaryl, heteroarylCι_4alkyl, heterocychc, or heterocyclicCι_4alkyl, wherein the aryl, heteroaryl and heterocychc rings may all be optionally substituted; Rd is NRgR , alkyl, arylCl-4alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-Cι_4alkyl, heteroarylC2-4 alkenyl, heterocychc, heterocyclicCι.4 alkyl, wherein the aryl, heteoaryl and heterocychc rings may all be optionally substituted.
DETAILED DESCRIPTION OF THE INVENTION
The compounds of Formula (I) may also be used in association with the veterinary treatment of mammals, other than humans, in need of inhibition of IL-8 or other chemokines which bind to the IL-8RA and RB receptors. Chemokine mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section. In compounds of Formula (I), suitably Ri is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Cj-io alkyl, such as CF3; Ci-io alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C2-10 alkenyl; Ci-io alkoxy, such as methoxy, or ethoxy; halosubstituted Ci-io alkoxy, such as trifluoromethoxy; azide; (CR Rg)q S(O)tR4, wherein t is 0, 1 or 2; hydroxy; hydroxy Ci-4alkyl, such as methanol or ethanol; aryl, such as phenyl or naphthyl; aryl Ci-4 alkyl, such as benzyl; aryloxy. such as phenoxy; aryl C1.4 alkyloxy, such as benzyloxy; heteroaryl; heteroarylalkyl heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl ; heteroaryl C2-10 alkenyl; heterocychc C2-10 alkenyl; (CRgR8)qNR4R5; C2-10 alkenyl C(O)NR4Rs; (CRgR8)q C(O)NR4R5; (CR8R8)q C(O)NR4Rlθ; S(O)3H; S(O)3R8; (CRgR8)q C(O)Rn; C2-IO alkenyl C(O)Rn; C2-10 alkenyl C(O)ORn; C(O)Rn; (CR8R8)q C(O)ORi2; (CRgRg)qOC(O)Rι 1; (CR8R8)q NR4C(O)Rι 1, (CR8R8)q NHS(O)2Rπ, (CR8R8)qS(O)2NR4R5; or two Ri moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; and s is an integer having a value of 1 to 3. The aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl. heterocychc, heterocyclicalkyl, and heterocyclicalkenyl moieties may all be optionally substituted as defined herein below.
Suitably, q is 0, or an integer having a value of 1 to 10. When Ri forms a dioxybridge, s is preferably 1. When Rl forms an additional unsaturated ring, it is preferably 6 membered resulting in a naphthylene ring system. This naphthylene ring may be substituted independently, 1 to 3 times by the other R moieties as defined above.
Suitably, R4 and R5 are independently hydrogen, optionally substituted Cι_4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C i -4-dkyl, heterocychc, heterocyclicCi-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from O/N/S. R8 is suitably independently selected from hydrogen or C 1.4 alkyl.
RlO is suitably -io alkyl C(O)2R8, such as CH2C(O)2H or
CH2C(O)2CH3.
Rl 1 is suitably hydrogen, C1.4 alkyl, aryl, aryl Cι_4 alkyl, heteroaryl, heteroaryl Ci-4alkyl, heterocychc, or heterocychc Cι_4alkyl. R12 is suitably hydrogen, Cl-10 alkyl, optionally substituted aryl or optionally substituted arylalkyl.
Rl7 is suitably Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylCi-4alkyl, heterocychc, or heterocyclicCi-4alkyl, wherein the aryl, heteroaryl and heterocychc rings may all be optionally substituted. Preferably Ri is halogen, cyano. nitro, CF3, C(O)NR4R5, alkenyl
C(O)NR4R5, C(O) R4R10, alkenyl C(O)ORi2, heteroaryl, heteroarylalkyl , heteroaryl alkenyl. or S(O)NR4.R5, and preferably R4 and R5 are both hydrogen or one is phenyl. A preferred ring substitution for Ri is in the 4-position of the phenyl ring. In compounds of Formula (I), suitably R13 and R 14 are independently hydrogen, optionally substituted Cι_4 alkyl which may be straight or branched as defined herein, or one of R13 and R 14 are an optionally substituted aryl; v is an integer having a value of 1 to 4.
When R13 or R 4 are an optionally substituted alkyl, the alkyl moiety may be substituted one to three times independently by halogen; halosubstituted Ci_4 alkyl such as trifluromethyl; hydroxy; hydroxy Cι_4alkyl, Ci-4 alkoxy; such as methoxy, or ethoxy. halosubstituted Ci-io alkoxy, S(O)tR4; aryl; NR4R5; NHC(O)R4; C(O)NR4R5; or C(O)OR8. Suitably, Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR8R8)q S(O)tR4; hydroxy; hydroxyCi-4alkyl: aryl; aryl Ci-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocychc, heterocychc Cι_4alkyl; aryl C2-10 alkenyl; heteroaryl C2-IO alkenyl; heterocychc C2-10 alkenyl; (CR8Rg)q NR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)q C(O)NR4R5; (CR8R8)q C(O)NR4Rlθ; S(O) H; S(O)3Rg; (CR8R8)q C(O)Rn; C2-10 alkenyl C(O)Rn; C2-IO alkenyl C(O ORn; (CR8R8)q C(O)ORι2; (CR8Rg)q OC(O) Rn; (CRgRg)q NR4C(O)R 11 , (CR8R8)q NHS(O)2, (CR8R8)q S(O)2NR4R5 or two Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring When Y forms a dioxybridge, s is preferably 1. When Y forms an additional unsaturated ring, it is preferably 6 membered resulting in a naphthylene ring system. This naphthylene ring may be substituted 1 to 3 times by other Y moieties as defined above. The aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl. heterocychc, heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.
Rd is a NR5R7, alkyl, aryl C1.4 alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-C 1.4 alkyl, heteroarylC2-4 alkenyl, heterocychc, heterocyclicCι.4 alkyl, or heterocychc C 2-4 alkenyl moiety, wherein the aryl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heterocychc, and heterocyclicalkyl, and heterocyclicalkenyl moieties noted above may all be optionally substituted as defined herein.
Y is preferably a halogen, Ci-4 alkoxy, optionally substituted aryl, optionally substituted aryloxy or arylalkoxy, methylene dioxy, NR4R5, thio Cι_ 4alkyl, thioaryl, halosubstituted alkoxy, optionally substituted C1-.4 alkyl, or hydroxy alkyl. Y is more preferably mono-substituted halogen, disubstituted halogen, mono-substituted alkoxy, disubstituted alkoxy. methylenedioxy, aryl, or alkyl, more preferably these groups are mono or di-substituted in the 2'- position or
2'-, 3'-position.
While Y may be substituted in any of the 5 ring positions, preferably when R is OH, SH, or NHSO2Rd» is preferably mono-substituted in the 2'-position or 3 - position, with the 4'- preferably being unsubstituted. If the ring is disubstituted, when R is OH, SH, or NSO2R , substituents are preferably in the 2' or 3' position of a monocyclic ring. While both Ri and Y can both be hydrogen, it is preferred that at least one of the rings be substituted, and more preferably that both rings are substituted. Exemplified compounds of Formula (I) include:
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-methoxy guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N",N"-diisopropyl guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-piperidinyl guanidine; N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N",N"-pyrrolidinyl guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N",N"-diethyl guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-phenyl guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-[(3-hydroxymethyl)piperidinyl] guanidine; N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-(2- dimethylaminoethyl) guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-aminomethyl guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-[(+/-)-2-(piperidinomethyl)- piperidyl] guanidine;
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-[(+/-)-2-(dimethylaminomethyl)- piperidyl] guanidine; and
N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-(3- dimethylaminopropyl) guanidine. As used herein, "optionally substituted" unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted Ci-ioalkyl; Ci-io alkoxy, such as methoxy or ethoxy; S(O)m' Ci-io alkyl, wherein m' is 0, 1 or 2, such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono & di-substituted amino, such as in the NR4R5 group; NHC(O)R4; C(O)NR4R5; C(O)OH; S(O)2NR4Rs; NHS(O)2Rl5, Ci-io alkyl, such as methyl, ethyl, propyl, isopropyl, or t-butyl; halosubstituted Ci-io alkyl, such as CF3; an optionally substituted aryl, such as phenyl, or an optionally substituted arylalkyl, such as benzyl or phenethyl, optionally substituted heterocylic, optionally substituted heterocy lie alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl alkyl, wherein these aryl , hetroaryl, or heterocychc moieties may be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; Ci-io alkoxy; S(O)mCi-io alkyl; amino, mono & di-substituted amino, such as in the NR4R5 group; C i-io alkyl, or halosubstituted Ci- 10 alkyl, such as CF3. R15 is suitably Cι_4 alkyl, aryl, aryl Ci-4alkyl, heteroaryl, heteroarylCi-
4-tlkyl, heterocychc, or heterocyclicCi-4alkyl.
Another aspect of the present invention are the novel compounds of Formula (II), or a pharmaceutically acceptable salt thereof, as described below, which are also useful in inhibiting the binding of IL-8 to its receptors in a mammal in need thereof. This invention also relates to the pharmaceutical compositions comprising a compound of Formula (II) and a pharmaceutically acceptable diluent or carrier. Compounds of Formula (II) are also useful for treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8RA or RB receptor and which method comprises administering an effective amount of a compound of Formula (II) or a pharmaceutically acceptable salt thereof.
Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of inorganic and organic acids, such as hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane sulphonic acid, ethane sulphonic 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 mandelic acid. In addition, pharmaceutically acceptable salts of compounds of Formula (I) may also be formed with a pharmaceutically acceptable cation, for instance, if a substituent group comprises a carboxy moiety. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations.
The following terms, as used herein, refer to:
• "halo" - all halogens, that is chloro, fluoro, bromo and iodo. • "Cι_ιoalkyl" or "alkyl" - both straight and branched chain alkyls of 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, iso-butyl, tert-butyl, n- pentyl and the like.
• The term "cycloalkyl" is used herein to mean cyclic alkyls, preferably of 3 to 8 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl, and the like.
• The term "alkenyl" is used herein at all occurrences to mean straight or branched chain alkyls of 2-10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-l- propenyl, 1-butenyl, 2-butenyl and the like.
• "aryl" - phenyl and naphthyl;
• "heteroaryl" (on its own or in any combination, such as "heteroaryloxy", or "heteroaryl alkyl") - 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, thiazole, thiadiazole, triazole, imidazole, or benzimidazole.
• "heterocychc" (on its own or in any combination, such as "heterocyclicalkyl") - 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; such as, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, or imidazolidine.
• The term "arylalkyl" or "heteroarylalkyl" or "heterocyclicalkyl" is used herein to mean Ci-io alkyl, as defined above, attached to an aryl. heteroaryl or heterocychc moiety, as also defined herein, unless otherwise indicated.
• "sulfinyl" - the oxide S (O) of the corresponding sulfide. the term "thio" refers to the sulfide, and the term "sulfonyl" refers to the fully oxidized S(O)2 moiety.
• The term "wherein two Rl moieties (or two Y moieties) may together form a 5 or 6 membered unsaturated ring" is used herein to mean the formation of a napthylene ring system or a phenyl moiety having attached a 6 membered partially unsaturated ring such as a C cycloalkenyl, i.e hexene, or a C5 cyloalkenyl moiety, cyclopentene.
Methods of Preparation The compounds of Formula (I) may be obtained by applying synthetic procedures, some of which are illustrated in the Schemes below. The synthesis provided for in these Schemes is applicable for the producing compounds of Formula (I) having a variety of different R, Rl, and aryl groups which are reacted, employing optional substituents, which are suitably protected, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, in those cases, then affords compounds of the nature generally disclosed. Once the guanidine nucleus has been established, further compounds of these formulas may be prepared by applying standard techniques for functional group interconversion, well known in the art. While the schemes are shown with compounds only of Formula (I) this is merely for illustration purposes only.
Scheme 1
Figure imgf000014_0001
a) Br2, NaOAc, HOAc; b) CuCN, DMF, reflux; c) (BOC)20, DMAP, TEA; d) K2C03, MeOH; e) TFA
The desired aniline 6-scheme- 1 can be prepared from the commercially available benzoxazolinone 1 -scheme- 1. Bromide 2-scheme- 1 can be prepared from benzoxazolinone 1 -scheme- 1 using standard bromination conditions such as bromine and sodium acetate in acetic acid. Bromide 2-scheme- 1 can be converted to the cyanide 3-scheme-l using standard procedures such as copper (I) cyanide in refluxing DMF. The amide 3-scheme-l can be converted to the BOC protected compound 4-scheme-l using standard conditions such as BOC anhydride and triethylamine with a catalytic amount of dimethylaminopyridine in methylene chloride or another suitable organic solvent. The oxazolinone 4-scheme- 1 can be converted to the desired aniline 6-scheme- 1 by first hydrolysis to the phenol 5; scheme- 1 using standard conditions such as potassium carbonate in methanol followed by removal of the BOC protecting group using standard conditions such as trifluoroacetic acid in methylene chloride or another suitable organic solvent to give the aniline 6-scheme- 1. Scheme 2
Figure imgf000015_0001
a) RNCS; b) TBSCl; c) thiophosgene; d) RNH2
The desired thiourea 3-scheme-2 can be prepared as outlined in scheme 2.
The aniline 1 -scheme-2 can be coupled with a commercially available isothiocyanate or with an isothiocyanate made from condensing a commercially available amine with thiophosgene or a thiophosgene equivalent to give the thiourea 2-scheme-2. The phenol 2-scheme-2 can be protected as the TBS ether 3-scheme-2 using standard conditions such as TBSCl and imidazole in THF or another suitabel organic solvent. Alternatively the amine 1 -scheme-2 can be converted to the desired isothiocyanate by first protecting the phenol with a suitable protecting group such as the TBS ether 4-scheme-2 using standard conditions such as TBSCl and an amine base such as imidazole in THF or another suitable organic solvent. The thiourea 5 scheme-2 can be made by condensing the amine 4-scheme 2 with thiophosgene in the presence of base such as potassium carbonate. The desired thiourea 3-scheme-2 can be prepared from the isothiocyanate 5-scheme-2 by condensing it with the desired amine in a suitable organic solvent such as ethanol or DMF. Scheme 3
Figure imgf000016_0001
OTBS OTBS
b
Figure imgf000016_0002
a) MsCI, TEA; b) RRNH; c) TBAF, THF
The desired tetraalkylguanidine 4-scheme-3 can be prepared as outlined in scheme 3. The carbodiimide 2-scheme-3 can be prepared from the thiourea L scheme-3 under suitable conditions such as using an excess amount of methane sulfonyl chloride and a suitable amine base such as triethylamine in an organic solvent preferably methylene chloride at room temperature. The guanidine compound 3-scheme-3 can be obtained by utilizing standard procedures such as condensing the carbodiimide 2-scheme-3 and the desired secondary amine in a suitable solvent such as acetonitrile or THF at room temperature. The desired guanidine 4-scheme-3 can be obtained from 3-scheme-3 by removal of the TBS protecting group using standard conditions such as TBAF in THF under low reaction temperatures.
SYNTHETIC EXAMPLES The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. All temperatures are given in degrees centigrade, all solvents are highest available purity and all reactions run under anhydrous conditions in an argon atmosphere unless otherwise indicated.
In the Examples, all temperatures are in degrees Centigrade (°C). Mass spectra were performed upon a VG Zab mass spectrometer using fast atom bombardment, unless otherwise indicated. *H-NMR (hereinafter "NMR") spectra were recorded at 250 MHz using a Bruker AM 250 or Am 400 spectrometer. Multiplicities indicated are: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet and br indicates a broad signal. Sat. indicates a saturated solution, eq indicates the proportion of a molar equivalent of reagent relative to the principal reactant.
Example 1 Preparation of N-(4-cvano-2-hvdroxypheny-)-N'-(2-bromophenyl)-N"-methyl- N"-methoxy guanidine a) Preparation of 4-bromo- 1 ,2-benzoxazolinone . To a solution of benzoxazolinone (10 g, 74 mmol) in acetic acid (50 mL) at
0°C was added sodium acetate (7.4 g, 74 mmol) and bromine (3.8 mL, 74 mmol) and the reaction mixture was allowed to warm to rt. After 21 h, the precipitate was collected by filtration and washed with water. The filtrate was concentrated under reduced pressure to give more solid material which was collected by filtration followed by washing with water. The material was combined to give 14 g (88%) of 4-bromo- 1 ,2-benzoxazolinone as a yellow solid which required no further purification. !H NMR (400 MHz, DMSO) δ 7.6 (s, 1H), 7.3 (d, 1H), 7.0 (d, 1H); MS(EI) m/e 212 (M+). b) Preparation of 4-cyano-l ,2-benzoxazolinone. To a solution of 4-bromo- 1 ,2-benzoxazolinone (5.0 g, 23 mmol) in DMF (11 mL) was added copper (I) cyanide (3.6 g, 39 mmol) and the reaction heated at 165°C. After 6.5 h. the reaction mixture was cooled to rt and water (20 mL) and sodium cyanide (3.6 g) were added and the reaction heated at 100°C. After 12 h, the reaction mixture was extracted with ethyl acetate and the combined organic layers were filtered through a scrub pad of silica gel eluting with ethyl acetate. The filtrate was concentrated under reduced pressure to give 1.9 g (51%) of 4-cyano-l,2- benzoxazolinone as brown solid which required no further purification. ^H NMR (400 MHz, DMSO) δ 7.8 (s, 1H), 7.6 (d, 1H), 7.2 (d, 1H). c) Preparation of N-t-butylacetoxy-4-cyano-l,2-benzoxazolinone. To a solution of 4-cyano-l ,2-benzoxazolinone (1.9 g, 15 mmol) in THF (50 mL) at 0°C was added triethylamine (2.5 mL, 18 mmol), DMAP (0.37 g, 3.0 mmol) and BOC anhydride (4.3 g, 20 mmol) and the reaction mixture was allowed to warm to rt. After 1.5 h, the mixture was quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give 4.3 g (100%) of N-t-butylacetoxy-4-cyano- 1 ,2-benzoxazolinone as a yellow solid which required no further purification. *H NMR (400 MHz, CDC13) δ 7.8 (d, 1H), 7.55 (d, lH), 7.45 (s, 1H), 1.65 (s, 9H). d) Preparation of N-t-butylacetoxy-4-cyano-2-hydroxyl aniline. To a solution of N-t-butylacetoxy-4-cyano-l,2-benzoxazolinone (4.3 g, 16 mmol) in methanol (50 mL) was added potassium carbonate (2.3 g, 16 mmol). After 1.5 h, the reaction was quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under educed pressure to give 3.1 g (81%) of N-t-butylacetoxy-4-cyano-2-hydroxyl aniline as a brown foam which required no further purification. lU NMR (400 MHz, CDCI3) δ 7.7 (d, 1H), 7.15 (s, 1H), 7.1 (d,
1H), 1.5 (s, 9H). e) Preparation of 4-cyano-2-hydroxyl aniline.
To a solution of N-t-butylacetoxy-4-cyano-2-hydroxyl aniline (3.1 g, 13 mmol) in methylene chloride (100 mL) at 0°C was added trifluoroacetic acid and the reaction mixture was allowed to warm to rt. After 2.5 h, the reaction was quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude material was purified by flash column chromatography (50% ethyl acetate/hexanes) to give 1.7 g (96%) of 4-cyano-2- hydroxyl aniline as a tan solid. JH NMR (400 MHz. DMSO) δ 7.0 (d, IH), 6.85 (s, IH), 6.65 (d, IH); MS(EI) m/e 134 (M+). f) Preparation of N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl) thiourea.
To a solution of 4-cyano-2-hydroxyl aniline (1.0 g, 7.5 mmol) in ethanol (20 mL) was added 2-bromophenylisothiocyanate ( 1.0 mL, 7.5 mmol). After 24 h, the reaction mixture was concentrated under reduced pressure to give 2.0 g (77%) of N- (4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl) thiourea as a yellow solid which required no further purification. 1H NMR (400 MHz, DMSO) δ 10.8 (s, IH), 10.1 (s, IH), 9.6 (s, IH), 8.7 (d, IH), 7.7 (d, IH), 7.6 (d, IH), 7.4 (t, IH), 7.25 (d, IH), 7.2 (s, IH and d, 2H); MS(EI) m/e 229 ( 100), 348 (75 (M+)), 462 (30), 695 (10). g) Preparation of N-(4-cyano-2-t-butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) thio urea.
To a solution of N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl) thio urea (3.5 g, 10 mmol) in THF (50 mL) at 0°C was added imidazole (1.0 g, 15 mmol) and TBSCl (1.5 g, 10 mmol). After 1 h, the reaction mixture quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude material was crystallized from ethyl acetate to give 3.9 g (84%) of N-(4-cyano-2-t-butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) thio urea as a yellow solid. lH NMR (400 MHz, DMSO) δ 9.9 (s, IH), 9.2 (s, IH), 8.1 (d, IH), 7.7 (d, IH), 7.6 (d, IH), 7.45 (d, IH), 7.4 (t, IH), 7.35 (s, IH), 7.2 (t, IH); MS(EI) m/e 347 (100 (M+)), 175 (40), 461 (40). h) Preparation of N-(4-cyano-2-t-butyldimethylsilanoxyphenyl)-N'-(2- bromophenyl) carbodiimide. To a solution of N-(4-cyano-2-t-butyldimethylsilanoxyphenyl)-N'-(2- bromophenyl) thio urea (3.4 g, 7.4 mmol) in methylene chloride (40 mL) at 0°C was added triethylamine (3.1 mL, 22 mmol), DMAP (20 mg) and methane sulfonyl chloride (1.1 mL, 15 mmol). After 25 min, the reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give 3.3 g (100%) of N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide as a yellow solid which required no further purification. *H NMR (400 MHz, DMSO) δ 1.1 (d, IH), 7.45 (d, IH), 7.4 (m, 4H), 7.15 (t, IH). i) General procedure for the preparation of tetraalkylguandines. Preparation of N- (4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-methoxy guanidine.
To a solution of N-(4-cyano-2-t-butyldimethylsilanoxyphenyl)-N'-(2- bromophenyl) carbodiimide (96 mg, 0.22 mmol) in acetonitrile (2 mL) was added diisopropylmethyl amine (54 μL, 0.48 mmol) and methoxylmethylamine hydrochloride (23 mg, 0.24 mmol). After 30 min, the reaction was concentrated under reduced pressure. The crude material was diluted with THF (2.5 mL) and methanol (0.1 mL) and TBAF (0.3 mL, 0.26 mmol) were added at 0°C. After 45 min, the reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude residue was purified by recrystalization from methylene chloride and hexanes to give 58 mg (70%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"- methoxy guanidine as a white powder. ^H NMR (400 MHz, MeOD) δ 7.9 (s, IH), 7.45 (d, IH), 7.15 (t, IH), 7.05 (m, 2H), 7.0 (s, IH), 6.8 (t, IH); MS(EI) m/e 376 (100 (M+)), 186 (70).
Example 2 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-N",N"- diisopropyl guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (0.11 g, 0.26 mmol), diisopropylamine (41 μL, 0.29 mmol) and TBAF (0.3 mL, 0.29 mmol) in THF (3 mL) to give 30 mg (28%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2- bromophenyl)-N",N"-diisopropyl guanidine as a white solid. ^H NMR (400 MHz, MeOD) δ 7.3 (d, IH), 6.9 (t, IH), 6.8-6.7 (m, 4H), 6.6 (t, IH); MS(EI) m/e 416 (100) (NT). Example 3 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N'-(2-bromophenvI)-piperidinyl guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (0.12 g, 0.27 mmol), piperidine (30 μL, 0.30 mmol), diisopropylmethyl amine (34 μL, 0.30 mmol) and TBAF (0.4 mL, 0.41 mmol) in THF (3 mL) to give 47 mg (43%) of N-(4-cyano-
2-hydroxyphenyl)-N'-(2-bromophenyl)-piperidinyl guanidine as a tan solid. ^H NMR (400 MHz, DMSO) δ 7.45 (d, IH), 7.15 (d, IH), 7.1 (d, IH), 6.95 (d, IH), 6.9 (d, IH), 6.85 (s, IH and t, IH); MS(EI) m/e 21 1 (100), 398 (40_(M+)), 289 (30), 332 (20).
Example 4 Preparation of N-(4-cvano-2-hydroxyphenvI)-N,-(2-bromophenyl)-N",N"- pyrrolidinyl guanidine. The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (0.11 g, 0.26 mmol), pyrrolidine (24 μL, 0.29 mmol), diisopropylmethyl amine (32 μL, 0.29 mmol) and TBAF (0.31 mL, 0.31 mmol) in THF (3 mL) to give 56 mg (56%) of N- (4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-pyrrolidinyl guanidine as a white solid. iH NMR (400 MHz, MeOD) δ 7.4 (d, IH), 7.1 (t, IH), 6.95 (m, 2H), 6.8 (s, IH and t, IH), 6.75 (d, IH); MS(EI) m/e 386 ( 100) (MN
Example 5 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N'-(2-bromophenyl)-N".N"- diethyl guanidine. The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (0.11 g, 0.26 mmol), diethylamine (30 μL, 0.29 mmol), diisopropylmethylamine (32 μL, 0.29 mmol) and TBAF (0.31 mL, 0.31 mmol) in THF (3 mL) to give 56 mg (56%) of N- (4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N",N"-diethyl guanidine as a white solid. ' R NMR (400 MHz, MeOD) δ 7.35 (d, IH), 7.0 (t, IH), 6.9-6.75 (m, 4H), 6.7 (t, IH), 3.4 (q, 4H), 1.2 (t, 6H); MS(EI) m/e 388 ( 100) (MN
Example 6 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-N",N"- dibenzyl guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (97 mg, 0.23 mmol), dibenzylamine (48 μL, 0.25 mmol), diisopropylmethylamine (28 μL, 0.25 mmol) and TBAF (0.28 mL, 0.28 mmol) in THF (3 mL) to give 75 mg (50%) of N- (4-cyano-2-hydroxyphenyl)-N'-(2-brornophenyl)-N",N"-dibenzyl guanidine as a white solid. iH NMR (400 MHz, MeOD) δ 7.4-7.2 (m, 1 IH), 7.05 (t, IH), 6.9-6.8 (m, 4H), 6.75 (t, IH), 4.55 (s, 4H); MS(EI) m/e 512 (100) (M+)-
Example 7 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N'-(2-bromophenyl)-N"-methvI- N"-phenyl guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (0.10 g, 0.24 mmol), methyl phenyl amine (28 μL, 0.26 mmol), diisopropylmethylamine (29 μL, 0.26 mmol) and TBAF (0.3 mL, 0.3 mmol) in THF (2.5 mL) to give 20 mg (20%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-N"-methyl-N"-phenyl guanidine as a white solid. *H NMR (400 MHz, MeOD) 67.4 (d, IH), 7.2 (m, 3H), 7.1-6.95 (m, 5H), 6.9 (d, IH), 6.85 (s, IH), 6.75 (d, IH); MS(EI) m/e 423 (100) (M+).
Example 8 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-f(3- hydroxymethvDpiperidinyll guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (79 mg, 0.18 mmol), 2-hydroxymethylpiperidine (23 mg, 0.20 mmol), diisopropylmethylamine (22 μL, 0.20 mmol) and TBAF (0.22 mL, 0.22 mmol) in THF (2 mL) to give 52 mg (67%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2-bromophenyl)-[(2- hydroxymethyl)piperidinyl] guanidine as a white solid. 1 H NMR (400 MHz, MeOD) δ 7.4 (d, IH), 7.1 (t, IH), 7.0 (d, IH), 6.9 (d, IH), 6.85-6.75 (m, 3H), 3.8 (d, IH), 3.65 (dt, IH), 3.4 (m, 2H), 3.05 (t, IH), 2.9 (t, IH), 1.8 (m, 2H), 1.65 (m, IH), 1.55 (m, IH), 1.25 (m, 2H).
Example 9 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-N"-methyl- N"-(2-dimethylaminoethyl) guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (88 mg, 0.21 mmol), N,N,N'-trimethylethylenediamine (29 μL, 0.23 mmol), diisopropylmethylamine (26 μL, 0.23 mmol) and TBAF (0.25 mL, 0.25 mmol) in THF (2 mL) to give 22 mg (25%) of N-(4-cyano-2-hydroxyphenyl)-N -(2- bromophenyl)-N"-methyl-N"-(2-dimethylaminoethyl) guanidine as a white solid. H NMR (400 MHz, MeOD) δ 7.4 (d, IH), 7.1-6.95 (m, 2H), 6.85 (d, IH), 6.8 (s, IH), 6,75 (t, IH), 3.6 (t, 2H), 3.0 (s, 3H), 2.8 (t, 2H), 2.45 (s, 6H).
Example 10 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-N"-methyl- N"-aminomethyl guanidine. The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (76 mg, 0.18 mmol), N,N'-dimethylhydrazine dihydrochloride (27 mg, 0.20 mmol), diisopropylmethylamine (67 μL, 0.59 mmol) and TBAF (0.22 mL, 0.22 mmol) in THF (2 mL) to give 46 mg (68%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2- bromophenyl)-N"-methyl-N"-aminomethyl guanidine as a white solid. ^H NMR
(400 MHz, MeOD) δ 7.4 (d, IH), 7.05 (m, 2H), 6.9 (d, IH), 6.85 (s, IH), 6.8 (d, IH), 6.75 (t, IH), 3.0 (s, 3H), 2.6 (s, 3H); MS(EI) m/e 375 (100) (M+). Example 11 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N,-(2-bromophenyl)-f(+/-)-2- (piperidinomethyl)-piperidyll guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (175 mg, 0.41 mmol), (+/-)-2-(piperidinomethyl)-piperidine (82 μL, 0.45 mmol), diisopropylmethylamine (51 μL, 0.45 mmol) and TBAF (0.50 mL, 0.49 mmol) in THF (4 mL) to give 79 mg (40%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2- bromophenyl)-[(+/-)-2-(piperidinomethyl)-piperidyl] guanidine as a white solid. 1H NMR (400 MHz, MeOD) δ 7.45 (d, IH), 7.1 (t, IH), 7.05 (d, 2H), 7.0 (d, IH), 6.9 (t, IH), 6.85 (s, IH), 6.75 (d, IH), 4.4 (m, IH), 3.55 (d, IH), 3.15 (m, 2H), 2.8 (m, 2H), 2.6 (m, 2H). 2.45 (d, IH), 1.7-1.35 (m, 11H); MS(EI) m/e 497 (100_(M*)), 248 (70), 270 (30).
Example 12 Preparation of N-f4-cvano-2-hvdroxyphenyl)-N'-(2-bromophenyl)-r(+/-)-2- (dimethylaminomethyl)-piperidvπ guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (68 mg, 0.16 mmol), (+/-)-N-(2-piperidylmethyl)-dimethyl amine (26 μL, 0.18 mmol), diisopropylmethylamine (20 μL. 0.18 mmol) and TBAF (0.20 mL. 0.19 mmol) in THF (2 mL) to give 22 mg (30%) of N-(4-cyano-2-hydroxyphenyl)-N'-(2- bromophenyl)-[(+/-)-2-(dimethylaminomethyl)-piperidyl] guanidine as a white solid.
*H NMR (400 MHz, MeOD) δ 7.5 (d, IH), 7.2 (t, IH), 7.15 (d, IH), 7.05 (d, IH), 6.95 (s, IH), 6.9 (m, 2H), 4.35 (m, IH), 3.75 (d, IH), 3.2 (m, 2H), 2.5 (s, 6H), 2.4 (d, IH), 1.95 (m, IH), 1.7 (m, 2H), 1.55 (m, 2H), 1.3 (m. 2H); MS(EI) m/e 457 (100 (M+)), 227 (75), 250 (50), 270 (45). Example 13 Preparation of N-(4-cvano-2-hvdroxyphenyl)-N>-(2-bromophenyl)-N"-methyl- N"-(3-dimethylaminopropyl) guanidine.
The standard procedure was followed using N-(4-cyano-2-t- butyldimethylsilanoxyphenyl)-N'-(2-bromophenyl) carbodiimide (65 mg, 0.15 mmol), N,N,N'-trimethyl- 1 ,3-diaminopropane (23 μL, 0.16 mmol), diisopropylmethylamine (18 μL, 0.16 mmol) and TBAF (0.18 mL, 0.18 mmol) in THF (2 mL) to give 46 mg (47 %) of N-(4-cyano-2-hydroxyphenyl)-N'-(2- bromophenyl)-N"-methyl-N"-(3-dimethylaminopropyl) guanidine as a white solid. !H NMR (400 MHz. MeOD) δ 7.35 (d, IH), 7.0 (t, IH), 6.85 (d, IH), 6.75 (m, 3H), 6.65 (d, IH), 3.5 (t, 2H), 2.85 (s, 3H), 2.4 (t, 2H), 2.2 (s, 6H), 1.85 (t, 2H); MS(EI) m/e 431 (100 (W)), 1 16 (70), 216 (55), 237 (40), 256 (10).
METHOD OF TREATMENT The compounds of Formula (I) and (II), or a pharmaceutically acceptable salt thereof can be used in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or other mammal, which is exacerbated or caused by excessive or unregulated IL-8 cytokine production by such mammal's cell, such as but not limited to monocytes and or macrophages, or other chemokines which bind to the IL-8 α or β receptor, also referred to as the type I or type II receptor.
Accordingly, the present invention provides a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8 α or β receptor and which method comprises administering an effective amount of a compound of Formula (I) or (II) or a pharmaceutically acceptable salt thereof. In particular, the chemokines are IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78.
For purposes herein, the compounds of Formula (I) and (II) all have the same dosages, and formulations as that of Formula (I) are used interchangeably.
The compounds of Formula (I) are administered in an amount sufficient to inhibit cytokine function, in particular IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78, such that they are biologically regulated down to normal levels of physiological function, or in some case to subnormal levels, so as to ameliorate the disease state. Abnormal levels of IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 for instance in the context of the present invention, constitute: (I) levels of free IL-8 greater than or equal to 1 picogram per mL; (ii) any cell associated IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 above normal physiological levels; or (iii) the presence EL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 above basal levels in cells or tissues in IL-8, GROα. GROβ, GROγ, NAP-2 or ENA-78 respectively, is produced.
There are many disease states in which excessive or unregulated IL-8 production is implicated in exacerbating and/or causing the disease. Chemokine mediated diseases include psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, stroke, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft vs. host reaction, alzheimers disease, allograft rejections, malaria, restinosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis or undesired hematopoietic stem cells release.
These diseases are primarily characterized by massive neutrophil infiltration, T-cell infiltration, or neovascular growth, and are associated with increased IL-8, GROα, GROβ, GROγ or NAP-2 production which is responsible for the chemotaxis of neutrophils into the inflammatory site or the directional growth of endothelial cells. In contrast to other inflammatory cytokines (IL-1, TNF, and IL-6), IL-8, GROα, GROβ, GROγ or NAP-2 has the unique property of promoting neutrophil chemotaxis, enzyme release including but not limited to elastase release as well as superoxide production and activation. The α-chemokines but particularly, GROα, GROβ, GROγ or NAP-2, working through the IL-8 type I or II receptor can promote the neovascularization of tumors by promoting the directional growth of endothelial cells. Therefore, the inhibition of IL-8 induced chemotaxis or activation would lead to a direct reduction in the neutrophil infiltration. Recent evidence also implicates the role of chemokines in the treatment of HIV infections, Littleman et al., Nature 381, pp 661 (1996) and Koup et al., Nature 381, pp 667 (1996).
The present invention also provides for a means of treating, in an acute setting, as well as preventing, in those individuals deemed susceptible to, CNS injuries by the chemokine receptor antagonist compounds of Formula (I).
CNS injuries as defined herein include both open or penetrating head trauma, such as by surgery, or a closed head trauma injury, such as by an injury to the head region. Also included within this definition is ischemic stroke, particularly to the brain area.
Ischemic stroke may be defined as a focal neurologic disorder that results from insufficient blood supply to a particular brain area, usually as a consequence of an embolus. thrombi, or local atheromatous closure of the blood vessel. The role of inflammatory cytokines in this area has been emerging and the present invention provides a mean for the potential treatment of these injuries. Relatively little treatment, for an acute injury such as these has been available.
TNF-α is a cytokine with proinflammatory actions, including endothelial leukocyte adhesion molecule expression. Leukocytes infiltrate into ischemic brain lesions and hence compounds which inhibit or decrease levels of TNF would be useful for treatment of ischemic brain injury. See Liu et al., Stroke, Vol. 25., No. 7, pp 1481-88 (1994) whose disclosure is incorporated herein by reference.
Models of closed head injuries and treatment with mixed 5-LO/CO agents is discussed in Shohami et al., J. of Vaisc & Clinical Physiology and Pharmacology, Vol. 3, No. 2, pp. 99-107 (1992) whose disclosure is incorporated herein by reference. Treatment which reduced edema formation was found to improve functional outcome in those animals treated.
The compounds of Formula (I) are administered in an amount sufficient to inhibit binding to the IL-8 alpha or beta receptors such as evidenced by a reduction in neutrophil chemotaxis and activation. The discovery that the compounds of Formula (I) are inhibitors of IL-8 binding is based upon the effects of the compounds of Formulas (I) in the in vitro receptor binding assays which are described herein. The compounds of Formula (I) have been shown, in some instances, to be dual inhibitors of both recombinant type I and type II IL-8 receptors. Preferably the compounds are inhibitors of only one receptor, more preferably Type II.
As used herein, the term "IL-8 mediated disease or disease state" refers to any and all disease states in which IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 plays a role, either by production of IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 themselves, or by IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by IL-8.
As used herein, the term "chemokine mediated disease or disease state" refers to any and all disease states in which a chemokine which binds to an IL-8 α or β receptor plays a role, such as but not limited IL-8, GROα. GROβ, GROγ, NAP-2 or ENA-78. This would include a disease state in which, IL-8 plays a role, either by production of IL-8 itself, or by IL-8 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by IL-8. As used herein, the term "cytokine" refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response. A cytokine includes, but is not limited to, monokines and lymphokines, regardless of which cells produce them. For instance, a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte. Many other cells however also produce monokines. such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes and B -lymphocytes. Lymphokines are generally referred to as being 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).
As used herein, the term "chemokine" refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response, similar to the term "cytokine" above. A 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, GRO-γ, NAP-2, ENA-78, IP- 10, MlP-lα, MlP-β, PF4, and MCP 1, 2, and 3.
In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy, it will normally be formulated into a pharmaceutical , composition in accordance with standard pharmaceutical practice. This 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.
Compounds of Formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation. The compounds of Formula (I) may be administered in conventional dosage forms prepared by combining a compound of Formula (I) with standard pharmaceutical carriers according to conventional procedures. The compounds of Formula (I) may also be administered in conventional dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation. It will be appreciated that the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. The carrier(s) 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 employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrier or diluent may include time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax. A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably will be from about 25mg. to about lg. When a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.
Compounds of Formula (I) may be administered topically, that is by non- systemic administration. This includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.
Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. The active ingredient may comprise, for topical administration, from 0.001% to 10% w/w, for instance from 1% to 2% by weight of the Formulation. It may however comprise as much as 10% w/w but preferably will comprise less than 5% w/w, more preferably from 0.1% to 1% w/w of the Formulation. Lotions according to the present invention include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those for the preparation of drops. Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.
Creams, ointments or pastes according to the present invention are semi-solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with the aid of suitable machinery, with a greasy or non-greasy base. The base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as steric or oleic acid together with an alcohol such as propylene glycol or a macrogel. The formulation may incorporate any suitable surface active agent such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.
Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and preferably including a surface active agent. The resulting solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98-100 °C. for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to the container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%). Suitable solvents for the preparation of an oily solution include glycerol. diluted alcohol and propylene glycol.
Compounds of formula (I) may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Appropriate dosage forms for such administration may be prepared by conventional techniques. Compounds of Formula (I) may also be administered by inhalation, that is by intranasal and oral inhalation administration. Appropriate dosage forms for such administration, such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.
For all methods of use disclosed herein for the compounds of Formula (I), the daily oral dosage regimen will preferably be from about 0.01 to about 80 mg/kg of total body weight. The daily parenteral dosage regimen about 0.001 to about 80 mg/kg of total body weight. The daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times daily. The daily inhalation dosage regimen will preferably be from about 0.01 mg/kg to about 1 mg/kg per day. It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula (I) or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.
The invention will now be described by reference to the following biological examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. BIOLOGICAL EXAMPLES
The IL-8, and Gro-α chemokine inhibitiory effects of compounds of the present invention were determined by the following in vitro assay: Receptor Binding Assays: [125l] IL-8 (human recombinant) was obtained from Amersham Corp.,
Arlington Heights, IL, with specific activity 2000 Ci/mmol. Gro-α was obtained from NEN- New England Nuclear. All other chemicals were of analytical grade. High levels of recombinant human IL-8 type α and β receptors were individually expressed in Chinese hamster ovary cells as described previously (Holmes, et al., Science, 1991, 253, 1278). The Chinese hamster ovary membranes were homogenized according to a previously described protocol (Haour, et al., J Biol Chem., 249 pp 2195-2205 (1974)). Except that the homogenization buffer was changed to lOmM Tris-HCL, lmM MgS04, 0.5mM EDTA (ethylene-diaminetetra- acetic acid), ImMPMSF (α-toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH 7.5. Membrane protein concentration was determined using Pierce Co. micro-assay kit using bovine serum albumin as a standard. All assays were performed in a 96-well micro plate format. Each reaction mixture contained 125τ iL_g (0.25 nM) or 125τ Gro-α and 0.5 μg/mL of IL-8Rα or 1.0 μg/mL of IL-8Rβ membranes in 20 mM Bis- Trispropane and 0.4 mM Tris HCl buffers, pH 8.0, containing 1.2 mM MgSO4, 0.1 mM EDTA, 25 mM NaCl and 0.03% CHAPS. In addition, drug or compound of interest was added which had been pre-dissolved in DMSO so as to reach a final concentration of between 0.0 InM and 100 uM. The assay was initiated by addition of 12-5I-IL-8. After 1 hour at room temperature the plate was harvested using a Tomtec 96-well harvester onto a glass fiber filtermat blocked with 1 % polyethylenimine/0.5% BSA and washed 3 times with 25 mM NaCl, 10 mM
TrisHCl, 1 mM MgSO4, 0.5 mM EDTA, 0.03 % CHAPS, pH 7.4. The filter was then dried and counted on the Betaplate liquid scintillation counter. The recombinant IL-8 Rα, or Type I, receptor is also referred to herein as the non- permissive receptor and the recombinant IL-8 Rβ, or Type II, receptor is referred to as the permissive receptor.
All of the exemplified compounds of Formulas (I) noted herein in the Synthetic Chemistry Section, Example 1 to 15, demonstrated an IC50 from about 45 to about <1 μg/mL in the permissive models for IL-8 receptor inhibition. Of those compounds tested, Examples 1 to 12 were also found to be inhibitors of Gro-α binding at about the same level. Chemotaxis Assay :
The in vitro inhibitory properties of these compounds are determined in the neutrophil chemotaxis assay as described in Current Protocols in Immunology, vol I, Suppl 1, Unit 6.12.3., whose disclosure is incorporated herein by reference in its entirety. Neutrophils where isolated from human blood as described in Current Protocols in Immunology Vol I, Suppl 1 Unit 7.23.1, whose disclosure is incorporated herein by reference in its entirety. The chemoattractants IL-8, GRO-α, GRO-β, GRO-γ and NAP-2 are placed in the bottom chamber of a 48 multiwell chamber (Neuro Probe, Cabin John, MD) at a concentration between 0.1 and 100 nM. The two chambers are separated by a 5um polycarbonate filter. When compounds of this invention are tested, they are mixed with the cells (0.001 - 1000 nM) just prior to the addition of the cells to the upper chamber. Incubation is allowed to proceed for between about 45 and 90 min at about 37°C in a humidified incubator with 5% CO2. At the end of the incubation period, the polycarbonate membrane is removed and the top side washed, the membrane then stained using the Diff Quick staining protocol (Baxter Products, McGaw Park, IL, USA). Cells which have chemotaxed to the chemokine are visually counted using a microscope. Generally, four fields are counted for each sample, these numbers are averaged to give the average number of cells which had migrated. Each sample is tested in triplicate and each compound repeated at least four times. To certain cells (positive control cells) no compound is added, these cells represent the maximum chemotactic response of the cells. In the case where a negative control (unstimulated) is desired, no chemokine is added to the bottom chamber. The difference between the positive control and the negative control represents the chemotactic activity of the cells. Elastase Release Assay:
The compounds of this invention are tested for their ability to prevent Elastase release from human neutrophils. 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 106 cells suspended in Ringer's Solution (NaCl 118, KC14.56, NaHCO3 25, KH2PO4 1.03, Glucose 11.1, HEPES 5 mM, pH 7.4) are placed in each well of a 96 well plate in a volume of 50 ul. To this plate is added the test compound (0.001 - 1000 nM) in a volume of 50 ul, Cytochalasin B in a volume of 50 ul (20ug/ml) and Ringers buffer in a volume of 50 ul. These cells are allowed to warm (37 °C, 5% CO2, 95% RH) for 5 min before IL-8, GROα, GROβ, GROγ or NAP-2 at a final concentration of 0.01 - 1000 nM was added. The reaction is allowed to proceed for 45 min before the 96 well plate is centrifuged (800 xg 5 min) and 100 ul of the supernatant removed. This suppernatant is 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 ug/ml dissolved in phosphate buffered saline. Immediately, the plate is placed in a fluorescent 96 well plate reader (Cytofluor 2350, Millipore, Bedford, MA) and data collected at 3 min intervals according to the method of Nakajima et al J. Biol Chem 254 4027 (1979). The amount of Elastase released from the PMNs is calculated by measuring the rate of MeOSuc-Ala-Ala-Pro-Val-AMC degradation. TNF-α in Traumatic Brain Injury Assay
The present assay provides for examination of the expression of tumor necrosis factor mRNA in specfic brain regions which follow experimentally induced lateral fluid-percussion traumatic brain injury (TBI) in rats. Adult Sprague-Dawley rats (n=42) were anesthetized with sodium pentobarbital (60 mg/kg, i.p.) and subjected to lateral fluid-percussion brain injury of moderate severity (2.4 atm.) centered over the left temporaparietal cortex (n=18), or "sham" treatment (anesthesia and surgery without injury, n=18). Animals are sacrificed by decapitation at 1, 6 and 24 hr. post injury, brains removed, and tissue samples of left (injured) parietal cortex (LC), corresponding area in the contralateral right cortex (RC), cortex adjacent to injured parietal cortex (LA), corresponding adjacent area in the right cortex (RA), left hippocampus (LH) and right hippocampus (RH) are prepared. Total RNA was isolated and Northern blot hybridization is performed and quantitated relative to an TNF-α positive control RNA (macrophage = 100%). A marked increase of TNF- α mRNA expression is observed in LH (104±17% of positive control, p < 0.05 compared with sham), LC (105±21%, p< 0.05) and LA (69±8%, p < 0.01) in the traumatized hemisphere 1 hr. following injury. An increased TNF- α mRNA expression is also observed in LH (46±8%, p < 0.05), LC (30±3%, p < 0.01) and LA (32±3%, p < 0.01) at 6 hr. which resolves by 24 hr. following injury. In the contralateral hemisphere, expression of TNF- α mRNA is increased in RH (46±2%, p < 0.01), RC (4±3%) and RA (22±8%) at 1 hr. and in RH (28±11%), RC (7±5%) and RA (26±6%, p < 0.05) at 6 hr. but not at 24 hr. following injury. In sham (surgery without injury) or naive animals, no consistent changes in expression of TNF- α mRNA are observed in any of the 6 brain areas in either hemisphere at any times. These results indicate that following parasagittal fluid-percussion brain injury, the temporal expression of TNF-α mRNA is altered in specific brain regions, including those of the non-traumatized hemisphere. Since TNF-α is able to induce nerve growth factor (NGF) and stimulate the release of other cytokines from activated astrocytes, this post-traumatic alteration in gene expression of TNF-α plays an important role in both the acute and regenerative response to CNS trauma. CNS Injury model for IL-β mRNA
This assay characterizes the regional expression of interleukin-lβ (IL-lβ) mRNA in specific brain regions following experimental lateral fluid-percussion traumatic brain injury (TBI) in rats. Adult Sprague-Dawley rats (n=42) are anesthetized with sodium pentobarbital (60 mg/kg, i.p.) and subjected to lateral fluid-percussion brain injury of moderate severity (2.4 atm.) centered over the left temporaparietal cortex (n=18), or "sham" treatment (anesthesia and surgery without injury). Animals are sacrificed at 1, 6 and 24 hr. post injury, brains removed, and tissue samples of left (injured) parietal cortex (LC), corresponding area in the contralateral right cortex (RC), cortex adjacent to injured parietal cortex (LA), corresponding adjacent area in the right cortex (RA), left hippocampus (LH) and right hippocampus (RH) are prepared. Total RNA is isolated and Northern blot hybridization was performed and the quantity of brain tissue EL- IB mRNA is presented as percent relative radioactivity of EL- IB positive macrophage RNA which was loaded on same gel. At 1 hr. following brain injury, a marked and significant increase in expression of IL-1 B mRNA is observed in LC (20.0±0.7% of positive control, n=6, p < 0.05 compared with sham animal), LH (24.5±0.9%, p < 0.05) and LA (21.5±3.1%, p < 0.05) in the injured hemisphere, which remained elevated up to 6 hr. post injury in the LC (4.0±0.4%, n=6, p < 0.05) and LH (5.0±1.3%, p < 0.05). In sham or naive animals, no expression of L- IB mRNA is observed in any of the respective brain areas. These results indicate that following TBI, the temporal expression of IL-1 B mRNA is regionally stimulated in specific brain regions. These regional changes in cytokines, such as EL- IB play a role in the post-traumatic.
All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth. The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims. Without further elaboration, it is believed that one skilled in the area can, using the preceding description, utilize the present invention to its fullest extent. Therefore the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows.

Claims

What is Claimed is:
1. A compound of the formula:
X
\ ° / (CR13R14)V— N N
Figure imgf000038_0001
(I) wherein:
R is OH, SH, NHSO2Rd
Rl is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Ci-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; (CR8R8)q S(O)tR4; hydroxy; hydroxy Ci-4alkyl; aryl; aryl C1.4 alkyl; aryloxy; aryl Ci-4 alkyloxy; heteroaryl; heteroarylalkyl; heterocychc, heterocychc Ci-4alkyl; heteroaryl Ci-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl C2-IO alkenyl; heterocychc C2-10 alkenyl; (CR R )qNR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)q C(O)NR4R5; (CR8R8)q C(O)NR4Rιo; S(O)3H; S(O)3R8; (CRg 8)q C(O)Rι 1; C2-10 alkenyl C(O)Rι 1; C2-IO alkenyl C(O)ORι l(CR8R8)q C(O)ORi2; (CR8R8)q OC(O) Ri 1;
(CR8R8)qNR4C(O)Rn, (CR8R8)q NHS(O)2Rι7, (CR8R8)q S(O)2NR4R5; or two Ri moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring; n is an integer having a value of 1 to 3; m is an integer having a value of 1 to 3; q is 0, or an integer having a value of 1 to 10; t is 0, or an integer having a value of 1 or 2; s is an integer having a value of 1 to 3; v is an integer having a value of 0 to 4; R4 and R5 are independently hydrogen, optionally substituted C 1.4 alkyl, optionally substituted aryl, optionally substituted aryl Ci-4-Ukyl, optionally substituted heteroaryl, optionally substituted heteroaryl Ci-4alkyl, heterocychc, heterocyclicC 1-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur; R6 and Rγ are independently hydrogen or a Ci-4 alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur;
Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted Ci-io alkyl; Cj-io alkyl; C2-10 alkenyl; Ci-io alkoxy; halosubstituted Ci-io alkoxy; azide; S(O)tR4; hydroxy; hydroxyCi-4alkyl; aryl; aryl Ci-4 alkyl; aryloxy; arylCi-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl Ci-4 alkyloxy; heterocychc, heterocychc Ci-4alkyl; aryl C2-10 alkenyl; heteroaryl
C2-10 alkenyl; heterocychc C2-10 alkenyl; NR4R5; C2-IO alkenyl C(O)NR4R5; C(O)NR4R5; C(O)NR4Rιo; S(O)3H; S(O)3R8; Ci-io alkyl C(O)Rn; C2-10 alkenyl C(O)Rι 1; C2-10 alkenyl C(O)ORι 1; C(O)Rι 1; C(O)ORi2; OC(O) Rl l; NR4C(O)Rι 1; or two Y moieties together may form O-(CH2)sO- or a 5 to 6 membered unsaturated ring;
R8 is hydrogen or Ci-4 alkyl;
RlO is Ci-io alkyl C(O)2R8;
Rl l is hydrogen, Ci-4 alkyl, optionally substituted aryl, optionally substituted aryl
Ci-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylCi-4alkyl, optionally substituted heterocychc, or optionally substituted heterocyclicC 1 -4-tlkyl; Rl2 is hydrogen, Ci-io alkyl, optionally substituted aryl or optionally substituted arylalkyl; Rl3 and R 14 are independently hydrogen, optionally substituted Cj_4 alkyl or one of R13 and R14 may be optionally substituted aryl; Rl7 is Ci-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylCi-4alkyl, heterocychc, or heterocyclicC ι_4alkyl, wherein the aryl, heteroaryl and heterocychc rings may all be optionally substituted;
Rd is NR5R7, alkyl, arylCi^alklyl, arylC 2-4 alkenyl, heteroaryl, hetroaryl-C ι_4 alkyl, heteroarylC2-4 alkenyl, heterocychc, heterocyclicC 1.4 alkyl, wherein the aryl, heteoaryl and heterocychc rings may all be optionally substituted; or a pharmaceutically acceptable salt thereof.
2. The compound according to Claim 1 wherein Ri is halogen, cyano, nitro, CF3, C(O)NR4R5, alkenyl C(O)NR4R5, C(O) R4R10, alkenyl C(O)ORi2, heteroaryl, heteroarylalkyl , heteroaryl alkenyl, or S(O)NR4R5.
3. The compound according to Claim 1 wherein Y is halogen, Ci-4 alkoxy, optionally substituted aryl, optionally substituted arylalkoxy, methylene dioxy,
NR4R5, thioCι_4alkyl, thioaryl, halosubstituted alkoxy, optionally substituted
Ci-4alkyl, hydroxy alkyl.
4. A pharmaceutical composition comprising an effective amount of a compound according to any of Claims 1 to 3, and a pharmaceutically acceptable carrier or diluent.
5. A method of treating a chemokine mediated disease state, wherein the chemokine binds to an IL-8 α or β receptor in a mammal, which comprises administering to said mammal an effective amount of a compound of the formula according to claim 1.
6. The method according to Claim 5 wherein the mammal is afflicted with a chemokine mediated disease selected from the group consisting of psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, stroke, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft vs. host reaction, alzheimers disease, allograft rejections, malaria, restinosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis or undesired hematopoietic stem cells release.
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EP1180025A1 (en) * 1999-05-28 2002-02-20 Smithkline Beecham Corporation Il-8 receptor antagonists
EP1180025A4 (en) * 1999-05-28 2002-08-28 Smithkline Beecham Corp Il-8 receptor antagonists
US6875759B1 (en) 1999-07-21 2005-04-05 Kadmus Pharmaceuticals Substituted guanidines and the use thereof
WO2001007029A3 (en) * 1999-07-21 2001-08-23 Kadmus Pharmaceuticals Inc Substituted guanidines and the use thereof
EP1413302A2 (en) * 1999-07-21 2004-04-28 Kadmus Pharmaceuticals, Inc. Substituted guanidines for the treatment of pain
EP1413302A3 (en) * 1999-07-21 2004-05-12 Kadmus Pharmaceuticals, Inc. Substituted guanidines for the treatment of pain
WO2001007029A2 (en) * 1999-07-21 2001-02-01 Kadmus Pharmaceuticals, Inc. Substituted guanidines and the use thereof
US8318175B2 (en) 2002-12-19 2012-11-27 New York University Method for treating amyloid disease
US9409146B2 (en) 2002-12-19 2016-08-09 New York University Method for treating amyloid disease
US7893089B2 (en) 2006-04-21 2011-02-22 GlaxoSmithKline, LLC IL-8 receptor antagonists
US8906382B2 (en) 2011-07-19 2014-12-09 New York University Method for treating amyloid disease
US9295719B2 (en) 2011-07-19 2016-03-29 New York University Method for treating amyloid disease
US9770496B2 (en) 2011-07-19 2017-09-26 New York University Method for treating amyloid disease
US9926353B2 (en) 2011-07-19 2018-03-27 New York University Immunotherapeutic modulation of amyloidogenic disease using non-fibrillogenic, non-amyloidogenic polymerized proteins and peptides
US11332506B2 (en) 2011-07-19 2022-05-17 New York University Immunotherapeutic modulation of amyloidogenic disease using non-fibrillogenic, non-amyloidogenic polymerized proteins and peptides

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