C3A RECEPTOR LIGANDS

Abstract: Novel C3A ligands are provided. Methods of using the present compunds to treat immune and inflammation disease are also provided.

Description: C3A RECEPTOR LIGANDS FIELD OF THE INVENTION The present invention relates to novel C3A receptor hgands, pharmaceutical compositions containing these compounds and methods of using the present compounds to treat inflammation BACKGROUND OF THE INVENTION Anaphylatoxins are 74-77 amino acid bioactive fragments of C5, C3 and C4 that are generated in vivo during complement activation Binding of the anaphylatoxins to specific cell surface receptors initiates and maintains the inflammatory process The fragments are believed to elicit mast cell and basophil degranulation with release of histamine, cytokines and other inflammatory mediators and induce smooth muscle cell contraction They are potent inflammatory mediators, inducing cellular degranulation, smooth muscle contraction arachidomc acid metabolism, cytoktne release, cellular chemotaxis See Gerard, C . and Gerard, N P (1994) Ann Rev Immunol 12, 775-808, Hugh, T E ( 1984) Springer Semin lmmunopathol 7, 193-219, Bitter-Suermann, D (1988) in The Complement System, Ed by K Rother & G Till, Springer Verlag, Heidelberg 367-395 The present fragments have been implicated in the pathogenesis of a number of inflammatory diseases See Vogt, W (1986) Complement 3, 177- 188, Morgan, B P ( 1994) European J Clin Investigation 24, 219-228 Studies have demonstrated the presence of a C3A receptor (C 3A-R) on guinea pig platelets, rat mast cells human neutrophils, eosinophils and platelets (Bitter-Suermann, D (1988) in The Complement System, Ed by K Rother & G Till, Springer Verlag, Heidelberg 367-395) A single class of high affinity C3A binding sites has been characterized on human neutrophils and differentiated U937 cells (Klos, A , Bank, S , Gietz, C , Bautsch, W , Kohl, J Burg, M and Kretzsch ar, T (1992) Biochemistry 31 , 1 1274-1 1282) Competition binding and functional desensitization studies are consistent with the presence of a receptor for C3A which is distinct from the C5A-R (Bitter-Suermann, D (1988) in The Complement System, Ed by K Rother & G Till, Springer Verlag, Heidelberg 367-395, Klos, A , Bank, S , Gietz, C , Bautsch, W , Kohl, J , Burg, M and Kretzschmar, T ( 1992) Biochemistry 31, 11274- 1 1282) However, there is evidence that C3A and C4A may bind to the same receptor as the two anaphylatoxins cross desensitize guinea pig lleal tissue (Hugh, T E (1984) Springer Semin lmmunopathol 7, 193-219, Bitter-Suermann, D (1988) in The Complement System, Ed by K Rother & G Till, Springer Verlag, Heidelberg 367-395), although other investigators using guinea pig macrophages indicate that there may be separate receptors (Murakami, Y , Yamamoto, T , Imamichi, T , Nagasawa, S (1993) Immunol Lett 36, 301-304) Functional activity of the C 3A-R is sensitive to pertussis toxin, consistent with the binding site being composed of a GPCR (Klos, A , Bank, S , Gietz, C , Bautsch, W , Kohl, J , Burg, M , and Kretzschmar, T ( 1992) Biochemistry 31 , 11274- 11282) ^ A complete understanding of the role of C3A in the pathogenesis of the inflammatory response has been hampered by the lack of the cloned receptor The present invention provides methods of using and functional characterization of human C3A receptor This same receptor was recently independently cloned from an HL-60 library by low-stnngency screening with a fMetLeuPhe receptor probe and, lacking functional data, 1" claimed to be an orphan receptor (AZ3B,8) Mouse L cells expressing AZ3B failed to bind and respond to the agonists examined, although C3A was not tested (Roghc, A , Prossnitz, E R , Cavanagh, S L , Pan, Z, Zou, A & Ye, R D (1996) Bwchimica et Biophystca Acta 1305, 39-43) The present invention discloses compounds that antagonize C3A receptor function ^ Clearly, there is a need for factors that mediate inflammation and their roles in dysfunction and disease There is a need, therefore, for identification and characteπzation of compounds which antagonize C3A receptor function, and which can play a role in preventing, ameliorating or correcting dysfunctions or diseases Thus, C3A hgands offer a unique approach towards the pharmacotherapy of immune ^ and inflammatory diseases such as rheumatoid arthritis, Alzheimer's disease, psoriasis, gout, multiple sclerosis, systemic lupus erythe atosus, glomerulonephπtis and adult respiratory distress syndrome SUMMARY OF THE INVENTION The present invention involves compounds represented by Formula (I) hereinbelow ^ and their use as C3A receptor hgands which are useful in the treatment of a variety of diseases associated with complement activation and/or increased levels of anaphylatoxins, including but not limited to rheumatoid arthritis, Alzheimer's disease, psoπasis, gout, multiple sclerosis, systemic lupus erythematosus, glomerulonephπtis and adult respiratory distress syndrome The present invention further provides methods for antagonizing C3A receptors in an -' V animal, including humans, which composes administeπng to an animal in need of treatment an effective amount of a compound of Formula (I), indicated hereinbelow DETAILED DESCRIPTION OF THE INVENTION 35 The compounds of the present invention are selected from Foonula (I) hereinbelow Formula (I) wherein: A represents Cj.4 alkylene, optionally substituted by C ] _4 alkyl or aryl; or A forms a 5-8 membered fused aliphatic ring with the adjacent phenyl ring; m is an integer from 1 to 3 ; each R] is independently selected form the group consisting of halo, Cι _4 alkyl, methanesulfonyl, alkoxy, nitrile, dimethylamine, methylenedioxy and CF3; R2 is selected from the group consisting of optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted phenyl, optionally substituted alkylsulfonyl, heteroarylsufonyl, optionally substituted phenylalkylaminocarbonyl, phenylaminocarbonyl, optionally substituted phenylalkyl, optionally substituted phenylsulfonyl, optionally substituted phenylalkylsulfonyl, optionally substituted naphthylalkylsulfonyl, heteroaryl carbonyl, heteroarylalkanoyl, optionally substituted alkylcarbonyl, optionally substituted phenylcarbonyl, and optionally substituted phenylalkylcarbonyl; wherein any substituents are selected from the group consisting of acyl, amide, C j .4 alkyl, halo, methylenedioxy, phenoxy and alkoxy; or R2 forms a 5-8 membered ring with A; or R2 forms a 4 to 7 membered ring with the carbon atom having the R4 substituent; R3 represents hydrogen or methyl; and R4 represents hydrogen or methyl. Preferably, A represents unsubstituted methylene. Preferably m is 2. Preferably, m represents a 3,5 substitution on the aryl ring. Preferably, R1 represents chloro. Preferably, R2 represents phenylpropionyl. Preferably, R3 represents hydrogen. Preferably, R4 represents methyl. More preferably, R2 represents 3,5 dichlorophenylsulfonyl. As used herein, "alkyl" refers to an optionally substituted hydrocarbon group joined together by single carbon-carbon bonds. The alkyl hydrocarbon group may be linear, branched or cyclic, saturated or unsaturated. Preferably, the group is linear. Preferably, the group is unsubstituted. Preferably, the group is saturated. 5 As used herein "cycloalkyl" refers to 3-7 membered carbocyclic rings. As used herein "heterocycloalkyl" refers to 4-7 membered heterocyclic rings containing 1 to 2 heteroatoms selected from N, O and S. As used herein, "aryl" refers to an optionally substituted aromatic group with at least one ring having a conjugated pi-electron system, containing up to two conjugated or 0 fused ring systems. "Aryl" includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted. A preferred aryl group is phenyl. As used herein "acyl" refers to alkylcarbonyl. The compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All 15 of these compounds and diastereomers are contemplated to be within the scope of the present invention. Preferred compounds in the present invention include: ( 1 ,2-Diphenylethylamino)acetylarginine, (+/-)-[ l-Indanylamino]acetylarginine, 0 ( 1 -Naphthylmethylamino)acetylarginine, [l-(2,2-Diphenylethylamino)]acetylarginine, [ 1 -( 1 ,2,3,4-Tetrahydroisoquinolyl)]acetylarginine, [ 1 -Decahydroquinolinyl]acetylarginine, [ 1 -( 1 ,2,3,4-Tetrahydronaphthylamino)]acetylarginine, 25 (+/-)- [2- Indanylaminojacetylarginine, [ 1 -(2-Phenethylamino)]acetylarginine, 2,3-Dichlorobenzylaminoacetylarginine, 3,5-Dichlorobenzylaminoacetylarginine, 2,6-Dichlorobenzylaminoacetylarginine, -' 3,4-Dimethylbenzylaminoacetylarginine, 4-Dimethylaminobenzylaminoacetylarginine, 3,4-Dichlorobenzylaminoacetylarginine, 3-Chlorobenzylaminoacetylarginine, 2,3-Dimethoxybenzylaminoacetylarginine, ^5 Benzylaminoacetylarginine, [ 1 -(3,3-Diphenylpropylamino)]aminoacetylarginine, [l-(2-(3,4-Dimethoxyphenyl))ethylamino]aminoacetylarginine, [ 1 -(4-benzylpiperidinyl)]aminoacetylarginine, [l-(3-Phenylpropylamino)]aminoacetylarginine, Dibenzylaminoaminoacetylarginine, [l-(2-(l-Naphthyl)ethylamino))]acetylarginine, [ 1 -(4-Phenylpiperidinyl)]acety larginine, [ 1 -(2-Phenethylamino)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(4-pyridinecarbonyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-piperonyl]acetylarginine, [4-Methylsulfonylbenzylamino-N-benzoyl]acetylarginine, [4-Methylsulfonylbenzylamino-N-(2-thiopheneacetyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(2-phenylacetyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(3,4-dichlorobenzoyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(2-phenoxypropionyl)]acetylarginine, t4-Methylsulfonylbenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(3-phenylpropionyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(3-(methoxycarbonyl)propionyl)]acetylarginine, [4-Methylsulfonylbenzylamino-N-(cyclohexanecarbonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(4-pyridinecarbonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-piperonyl]acetylarginine, [4-Trifuoromethylbenzylamino-N-benzoyl]acetylarginine, [4-Trifuoromethylbenzylamino-N-(2-thiopheneacetyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(2-phenylacetyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(3,4-dichlorobenzoyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(3-phenylpropionyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(3-(methoxycarbonyl)propionyl)]acetylarginine, [4-Trifuoromethylbenzylamino-N-(cyclohexanecarbonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3-phenylpropionyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(phenylsulfonyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(4-pyridinecarbonyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-piperonyl]acetylarginine, [3,5-Dimethoxybenzylamino-N-benzoyl]acetylarginine, [3,5-Dimethoxybenzylamino-N-(2-thiopheneacetyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(2-phenylacetyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(3,4-dichlorobenzoyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(3-phenylpropionyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(3-(methoxycarbonyl)propionyl)]acetylarginine, [3,5-Dimethoxybenzylamino-N-(cyclohexanecarbonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(4-pyridinecarbonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-piperonyl]acetylarginine, [3,4-Dichlorobenzylamino-N-benzoyl]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-thiopheneacetyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-phenylacetyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3,4-dichlorobenzoyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3-phenylpropionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3-(methoxycarbonyl)propionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(cyclohexanecarbonyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(4-pyridinecarbonyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-piperonyl]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-benzoyl]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(2-thiopheneacetyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(2-phenylacetyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(3,4-dichlorobenzoyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(3-phenylpropionyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(3-(methoxycarbonyl)propionyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(cyclohexanecarbonyl)]acetylarginine, [3-Iodobenzylamιno-N-(4-pyndιnecarbonyl)]acetylargιnιne, [3-Iodobenzylamιno-N-pιperonyl]acetylargιnιne, [3-Iodobenzylamιno-N-benzoyl]acetylargιnιne, [3-Iodobenzylamιno-N-(2-thιopheneacetyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(2-phenylacetyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(3,4-dιchlorobenzoyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(3-pyπdιnecarbonyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(2-(4-chlorophenoxy)acetyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(3-phenylpropιonyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(3-(methoxycarbonyl)propιonyl)]acetylargιnιne, [3-Iodobenzylamιno-N-(cyclohexanecarbonyl)]acetylargιnιne, [Benzylamιno-N-(4-pyπdιnecarbonyl)]acetylargιnιne, [Benzylamιno-N-pιperonyl]acetylargιnιne, [Benzylamino-N-benzoy ljacetylargimne, [Benzylamιno-N-(2-thιopheneacetyl)]acetylargιnιne, [Benzylamιno-N-(2-phenylacetyl)]acetylargmιne, [Benzylamιno-N-(3,4-dιchlorobenzoyl)]acetylargtnιne, [Benzylamιno-N-(3-pyndιnecarbonyl)]acetylargιnιne, [Benzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [Benzylamιno-N-(2-(4-chlorophenoxy)acetyl)]acetylargιnιne, [Benzylamιno-N-(3-phenylpropιonyl)]acetylargιnιne, [Benzylamιno-N-(3-(rnethoxycarbonyl)propιonyl)]acetylargιnιne, [Benzylamιno-N-(cyclohexanecarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-pyπdιnecarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)]acetylargιnιne [3,5-Dιchlorobenzylamιno-N-( phenylsulfonyl)]acetylargιnιne [3,5-Dιchlorobenzylamιno-N-(3-chlorophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-chlorophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(ιsopropylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(benzenesulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(5-chlorothιophene-2-sulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(8-quιnolιnesulfonyl)]acetylargιnιne, [3,5-Dichlorobenzylamino-N-((2-(l-naphthyl)ethyl)sulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3,4-dichlorophenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(3,4-dichlorophenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3,4-dichlorophenylsulfonyl)]acetylarginine, [Piperonylamino-N-(3,4-dichlorophenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3-chloro-2-methylphenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(3-chloro-2-methylphenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3-chloro-2-methylphenylsulfonyl)]acetylarginine, [Piperonylamino-N-(3-chloro-2-methylphenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(4-nitrophenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(4-nitrophenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(4-nitrophenylsulfonyl)]acetylarginine, [Piperonylamino-N-(4-nitrophenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(2-naphthylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(2-naphthylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-naphthylsulfonyl)]acetylarginine, [Piperonylamino-N-(2-naphthylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(2,5-dichloro-3-thiophenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(2,5-dichloro-3- thiophenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2,5-dichloro-3- thiophenylsulfonyl)]acetylarginine, [Piperonylamino-N-(2,5-dichloro-3- thiophenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3,5-trifluoromethylphenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(3,5-trifluoromethylphenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3,5-trifluoromethylphenylsulfonyl)]acetylarginine, [Piperonylamino-N-(3,5-trifluoromethylphenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(4,5-dibromo-2-thiophenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(4,5-dibromo-2-thiophenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(4,5-dibromo-2-thioρhenylsulfonyl)]acetylarginine, [Piperonylamino-N-(4,5-dibromo-2-thiophenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3,4-dimethoxyphenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(3,4-dimethoxyphenylsulfonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3,4-dimethoxyphenylsulfonyl)]acetylarginine, [Piperonylamino-N-(3,4-dimethoxyphenylsulfonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(4-t-butylphenylsulfonyl)]acetylarginine, [4-Trifluoromethylbenzylamino-N-(4-t-butylphenylsulfonyl)]acetylarginine, [3,4-Dιchlorobenzylamιno-N-(4-t-butylphenylsulfonyl)]acetylargιnιne, [Pιperonylamιno-N-(4-t-butylphenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-thιophenylsulfonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(2-thιophenylsulfonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-thιophenylsulfonyl)]acetylargιnιne, [Pιperonylamιno-N-(2-thιophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-tπfluoromethoxyphenylsulfonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(4-tπfluoromethoxyphenylsulfonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(4-tπfluoromethoxyphenylsulfonyl)]acetylargιnιne, [Pιperonylamιno-N-(4-tπfluoromethoxyphenylsulfonyl)]acetylargtntne, [3,5-Dιchlorobenzylamιno-N-(cyclohexylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(cyclohexylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(cyclohexylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-phenethylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(2-phenethylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-phenethylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2,4-dιchlorobenzylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(2,4-dιchlorobenzylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2,4-dιchlorobenzylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-acetylphenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(3-acetylphenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(3-acetylphenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-phenylphenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(2-phenylphenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-phenylphenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-phenoxyphenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(4-phenoxyphenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(4-phenoxyphenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-phenylphenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(4-phenylphenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(4-phenylphenylammocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(ιsopropylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(ιsopropylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(ιsopropylamιnocarbonyl)]acetylargιnme, [3,5-Dιchlorobenzylamιno-N-(phenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(phenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(phenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(benzylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(benzylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(benzylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-tπfluoromethylphenylamιnocarbonyl)]acetylargιnιne, [4-Tπfluoromethylbenzylamιno-N-(2-tπfluoromethylphenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-tπfluoromethylphenylamιnocarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-chlorophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-chlorophenylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(8-quιnohnylsulfonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(4-( l-methyhmιdazolyl)sulfonyl)]acetylargιnιne, (N-Methyl-3,4-dιchlorobenzylamιno)acetylargιnιne, [N-(n-Butyl)-3-ιodobenzylamιno]acetylargιnιne, 2-[(3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]propιonylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]acetyl-N- -methylargιnιne, [(+/-)-N-(3,5-dιchlorobenzyl)pιpecolyl]acetylargιnιne, and [(+/-)-N-(3,5-dιchlorobenzyl)ρrolyl]argιnιne More preferred compounds useful in the present invention include (3,5-Dιchlorobenzylamιno)acetylargιnιne, ( 1 ,2-Dιpheny lethylamιno)acety larginine, [1 -(2,2-Dιphenylethylamιno) jacetylargimne, [ 1 -( 1 ,2,3,4-Tetrahydroιsoquιnolyl)]acetylargιnιne, [ 1 -Decahydroquinohnyljacety larginine, [ 1 -( 1 ,2,3,4-Tetrahy dronaphthylamιno)]acetylargιnιne, 3,4-Dιmethylbenzylamιnoacetylargιnιne, 3,4-Dιchlorobenzylamιnoacetylargιnιne, 3-Chlorobenzylamιnoacetylargιnιne, 2,3-Dιmethoxybenzylamιnoacetylargιnιne, [l-(3,3-Dιphenylpropylamιno)]amιnoacetylargιnιne, [l-(2-(3,4-Dιmethoxyphenyl))ethylamιno]amιnoacetylargιnιne, [ l -(3-Phenylpropylamιno)]amιnoacetylargιnιne, Dibenzylaminoaminoacetylarginine, [3,4-Dιchlorobenzylamιno-N-(3-pyπdιnecarbonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-(4-chlorophenoxy)acetyl)]acetylargιnιne, [3,4-(Methylenedιoxy)benzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-pyπdιnecarbonyl)]acetylargιnιne, 3,5-Dιchlorobenzylamιnoacetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(4-phenoxyphenylamιnocarbonyl)]acetylargιnme, [3,5-Dιchlorobenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-phenoxypropιonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(2-(4-chlorophenoxy)acetyl)]acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)]acetylargιnιne, and (N-Methyl-3,4-dιchlorobenzylamιno)acetylargιnιne Even more preferred compounds useful in the present invention include [3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]acetylargιnιne, [3,4-Dιchlorobenzylamιno-N-(4-phenoxyphenylamιnocarbonyl)]acetylargιnιne ( [3,5-Dιchlorobenzylamιno-N-(3,4-dιchlorophenylamιnocarbonyl)]acetylargιnιne, (3,5-Dιchlorobenzylamιno)acetylargιnιne, [3,5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)]acetylargιnιne, and (N-Methyl-3,4-dιchlorobenzylamιno)acetylargιnιne An especially preferred compound in the present invention is [3,5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]acetylargιnιne The present compounds can also be formulated as pharmaceutically acceptable salts and complexes thereof Pharmaceutically acceptable salts are non-toxic salts in the amounts and concentrations at which they are administered Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate and quinate Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuπc acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaπc acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaπc acid, and quinic acid Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, chohne, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxyhc acid or phenol are present The present invention provides compounds of Formula (I) above which can be prepared using standard techniques An overall strategy for prepaπng preferred compounds described herein can be earned out as described in this section The examples which follow illustrate the synthesis of specific compounds Using the protocols descπbed herein as a model, one of ordinary skill in the art can readily produce other compounds of the present invention All reagents and solvents were obtained from commercial vendors Starting mateπals (e g , amines and epoxides) were synthesized using standard techniques and procedures The present invention provides compounds of formula (I) above which can be prepared using standard techniques An overall strategy for prepaπng preferred compounds described herein can be earned out as descπbed in this section The examples which follow illustrate the synthesis of specific compounds Using the protocols described herein as a model, one of ordinary skill in the art can readily produce other compounds of the present invention All reagents and solvents were obtained from commercial vendors Starting materials were synthesized using standard techniques and procedures The general classes of compounds 5 and 6 are prepared as follows Argimne derivative 1 (Pbf = 2,2,4,6,7-pentamethyldιhydrobenzofuran-5-sulfonyl) is coupled to Wang resin (200-400 mesh) using EDC/DMAP to give the resin-bound intermediate 2 N-terminal deprotection and subsequent coupling with bromoacetic acid affords the bromide 3 Displacement of the bromide with 3,5-dichlorobenzylamme yields 4, which upon acylation with 3-phenylpropιonyl chloride or the equivalent acid (RC0 2 H/EDC/HOBT), and TFA deprotection affords 5 Sulfonylation of the intermediate 4 (PhS0 2 Cl/pyndine/CH 2 Cl 2 ) and TFA deprotection affords the sulfonamide 6 Reaction of 4 with an isocyanate in 1.1 pyπdine CH C1 2 yields the urea 7 Resin bound 5, 6 and 7 are obtained as their TFA salts upon precipitation of the products in ether With appropriate manipulation and protection of any chemical functionality, synthesis of the remaining compounds of Formula (I) is accomplished by methods analogous to those above and to those described in the Experimental section. In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of humans and other mammals, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition. The present ligands can be administered by different routes including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, topical, transdermal, or transmucosal administration. For systemic administration, oral administration is preferred. For oral administration, for example, the compounds can be formulated into conventional oral dosage forms such as capsules, tablets and liquid preparations such as syrups, elixirs and concentrated drops. Alternatively, injection (parenteral administration) may be used, e.g., intramuscular, intravenous, intraperitoneal, and subcutaneous. For injection, the compounds of the invention are formulated in liquid solutions, preferably, in physiologically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution. In addition, the compounds may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms can also be produced. Systemic administration can also be by transmucosal or transdermal means For transmucosal or transdermal administration, penetrants appropriate to the barπer to be permeated are used in the formulation Such penetrants are generally known m the art, and include, for example, for transmucosal administration, bile salts and fusidic acid deπvatives In addition, detergents may be used to facilitate permeation Transmucosal administration, for example, may be through nasal sprays, rectal suppositories, or vaginal suppositories For topical administration, the compounds of the invention can be formulated into ointments, salves, gels, or creams, as is generally known in the art The amounts of various calcilytic compounds to be administered can be determined by standard procedures taking into account factors such as the compound IC50, EC50, the biological half-life of the compound, the age, size and weight of the patient, and the disease or disorder associated with the patient The importance of these and other factors to be considered are known to those of ordinary skill in the art Amounts administered also depend on the routes of administration and the degree of oral bioavailabihty For example, for compounds with low oral bioavailabihty, relatively higher doses will have to be administered Preferably the composition is in unit dosage form For oral application, for example, a tablet, or capsule may be administered, for nasal application, a metered aerosol dose may be administered, for transdermal application, a topical formulation or patch may be administered and for transmucosal delivery, a buccal patch may be administered In each case, dosing is such that the patient may administer a single dose Each dosage unit for oral administration contains suitably from 0 01 to 500 mg/Kg, and preferably from 0 1 to 50 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, calculated as the free base The daily dosage for parenteral, nasal, oral inhalation, transmucosal or transdermal routes contains suitably from 0 01 mg to 100 mg/Kg, of a compound of Formula(I) A topical formulation contains suitably 0 01 to 5 0% of a compound of Formula (I) The active ingredient may be administered from 1 to 6 times per day, preferably once, sufficient to exhibit the desired activity, as is readily apparent to one skilled in the art As used herein, "treatment" of a disease includes, but is not limited to prevention, retardation and prophylaxis of the disease Diseases and disorders which might be treated or prevented, include immune and inflammation-related diseases or disorders such as rheumatoid arthπtis, Alzheimer's disease, psoπasis, gout, multiple sclerosis, systemic lupus erythematosus, glomerulonephπtis and adult respiratory distress syndrome Composition of Formula (I) and their pharmaceutically acceptable salts which are active when given orally can be formulated as syrups, tablets, capsules and lozenges. A syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid carrier for example, ethanol, peanut oil. olive oil, glycerine or water with a flavoring or coloring agent. Where the composition is in the form of a tablet, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers in a hard gelatin capsule shell. Where the composition is in the form of a soft gelatin shell capsule any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils, and are incorporated in a soft gelatin capsule shell. Typical parenteral compositions consist of a solution or suspension of a compound or salt in a sterile aqueous or non-aqueous carrier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpyrrolidone, lecithin, arachis oil or sesame oil. Typical compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional propellant such as dichlorodifluoromethane or trichlorofluoromethane. A typical suppository formulation comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent, for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogs. Typical dermal and transdermal formulations comprise a conventional aqueous or non-aqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane. Preferably the composition is in unit dosage form, for example a tablet, capsule or metered aerosol dose, so that the patient may administer a single dose. No unacceptable toxological effects are expected when compounds of the present invention are administered in accordance with the present invention. The biological activity of the compounds of Formula (I) are demonstrated by the tests indicated hereinbelow. Stable Expression of C3A Receptor in RBL-2H3 Cells To prepare C3A receptor for expression in mammalian cells, a 1 6 kb cDNA fragment was obtained by PCR amplification that encompassed the entire C3A Receptor open reading frame This fragment was subcloned into Kpnl /Hind III sites of the mammalian expression vector, pCDN (Aiyar, N , et al (1994) Mol Cell Bio 131 , 75-86) Ohgonucleotide pπmers used for PCR amplification were 5 -GA AGT GGT ACC ATG GCG TC -3' and 5 - GC TCC AAG CTT TCA CAC AGT TG -3' (the translation start and stop codons are underlined) RBL-2H3 cells were electroporated with C3A in the pCDN mammalian expression vector (Aiyar, N , et al (1994) Mol Cell Bio 131 , 75-86), exactly as described (DeMartino, J A , et al (1994) J Biol Chem 269, 14446-14450) Individual G418 resistant (400 μg ml) colonies were isolated and expanded Clonal cell lines expressing C3A receptor, as determined by ability of the cell line to respond to C3A in a calcium mobilization assay, were chosen for further functional and binding studies Preparation of Membranes RBL-2H3 cells expressing the human C3A receptor (hC3AR) were cultured to confluency at 37° C in a humidified incubator with 5% C02/95% air, in Earls MEM supplemented with non-essential amino acids, 10% fetal calf serum and 400 μg/ml G418 Although this cell line is normally adherent, nonadherent cells are always present in cultures The nonadherent cells were adapted to grow in suspension Nonadherent cells from three T-150 flasks were centπfuged at 1,000 x g for 10 min and resuspended in 50-ml of the above medium in a 250 ml shake flask and over 7-10 days the cells were expanded to 2 5 1 in a spinner flask Cells were harvested by centπfugation, 1 ,000 x g for 10 min at 4° C, and membranes were isolated using a modification of the procedure of Ross et al , (1977) Briefly, the cell pellet was washed with PBS and resuspended in 30 ml of hypotonic membrane buffer (20 mM Tπs, pH 7 5, 2 mM MgCl 2 , 0 1 mM EDTA, 1 mM DTT, 1 mM phenylmethylsulfonyl fluoride, 1 μM leupeptin, 1 μM pepstatin A) and incubated on ice for 5 min The cell suspension was homogenized in 40 ml Dounce homogenizer and centπfuged at 1,000 x g for 15 min to remove nuclei and cellular debris Cell membranes were pelleted at 100,000 x g for 30 min at 4° C Membranes were resuspended in membrane buffer with 10% sucrose and layered over membrane buffer with 40% sucrose and centπfuged at 100,000 x g for 90 min at 4° C Membranes at the interface were isolated and collected by centπfugation at 100,000 x g for 30 mm The membrane pellet was resuspended in 5 0 ml of membrane buffer and ahquots stored at -80°C Protein concentration was quantified using the BCA protein assay reagent (Pierce, Rockford, IL) Scintillation Proximity Assay All assays are performed in a 96-well micro-titre plate format The 96-well plates (1450-401) are obtained from Wallac, Turku, Finland Human anaphylatoxin C3A was obtained from Advanced Research Technologies, San Diego, CA with Bolton-Hunter custom lodination being performed by NEN Research Products, Boston, MA with specific activity of 2200 Ci/mmol Wheatgerm agglutimn SPA (Scintillation Proximity Assay) beads were obtained from Amersham Corp , Arlington Heights, IL The binding buffer consists of 20 mM Bis-Tπspropane, 25 mM NaCl, 1 mM MgSθ4, 0 1 mM EDTA at pH 8 0 Each reaction mixture contains 1251 C3A (25 pM, obtained from NEN, Boston MA), wheatgerm agglutimn SPA beads (0 1 mg), 0 35 μgs of RBL-2H3 C3A receptor membranes (this may vary with quality of membrane preparation), 23 ug/ml BSA and 0 03% CHAPS in binding buffer The membranes were prebound to SPA beads for 30 minutes on ice while shaking The mixture of membranes and beads were centπfuged for three minutes at 2000 rpm The supernatant was removed and the pellet was resuspended to original volume in binding buffer with 50 ug/mL BSA Samples of interest were dissolved in neat DMSO to yield a 20X solution followed by a 1 1 mixture with H2O to yield a 10X, 50% DMSO working solution The order of addition was 10 uLs sample, 45 uLs membrane bound SPA beads followed by 45 uLs of radiolabled ligand in binding buffer containing 0 06% CHAPS The plates were covered with plate sealers from Dynex Technologies, Inc, and shaken for 20 minutes and incubated an additional 40 minutes at room temperature The plates were then centπfuged for three minutes at 2000 rpm followed by counting on the Wallac 1450 Micro Beta Plus Liquid Scintillation counter Calcium Functional Assays 7TM receptors which are expressed in HEK 293 cells have been shown to be coupled functionally to activation of PLC and calcium mobilization and/or cAMP stimulation or inhibition Basal calcium levels in the HEK 293 cells in receptor- transfected or vector control cells were observed to be in the normal, 100 nM to 200 nM, range HEK 293 cells expressing recombinant receptors are loaded with fura 2 and in a single day > 150 selected hgands or tissue/cell extracts are evaluated for agonist induced calcium mobilization Agonists presenting a calcium transient are tested in vector control cells to determine if the response is unique to the transfected cells expressing receptor Calcium mobilization: C3a-induced response in RBL-2H3 cells carrying C3a receptor: Bioassays: The functional activity of an antagonist of the C3a receptor is demonstrated using the C3a-mduced Ca2+ mobilization in RBL-2H3 cells stably expressing C3a (RBL-2H3- C3a) RBL-2H3-C3a Cell Culture Conditions: RBL-2H3-C3a cells were cultured to near confluence in T-150 flasks at 37° C in a humidified incubator with 5% Cθ2/95% air in Earls MEM with Earls salts (Gibco) supplemented with non-essential am o acids and L-glutamate, with 10% fetal calf serum (Gibco) and 400 ug/ml G418 (Gibco) Fluorescent Measurements-Calcium Mobilization: The functional assay used to assess antagonist activity of compounds was C3a- mduced calcium mobilization in intact RBL-2H3-C3a cells Cells were washed with 50 mM Tπs, pH 7 4 containing 1 mM EDTA The [Ca^+j, was estimated with the calcium fluorescent probe fura 2 (Grynkiewicz, et al , J Biol Chem , 1985, 260, 3440-3450) The media was aspirated from RBL-2H3-C3a cells that were near confluence in T-150 flasks then 40 ml in Krebs Ringer Hensilet containing 0 1 % BSA, 1 1 mM MgCl2 and 5 mM HEPES, pH 7 4 (buffer A) was added The diacetoxymethoxy ester of fura 2 (fura 2 / AM) was added at a concentration of 2 μM and incubated for 45 min at 37° C Buffer A was aspirated off the RBL-2H3-C3a cells and 40 ml of Buffer A was added to the cells and incubated for an additional 20 mm to allow complete hydrolysis of the entrapped ester Buffer A was aspirated and cells covered with ~5ml of Delbeccos Phosphate Buffered Saline with 1 mM EDTA (no calcium or magnesium) for 5 min at 37° C Buffer is aspirated off and 40 ml of buffer A added to the cells which were then mechanically detached from the flasks RBL-2H3-C3a cells were maintained at room temperature until used in the fluorescent assay which was performed within 3 hours The fluorescence of fura 2 containing cells was measured with a fluorometer designed by the Johnson Foundation Biomedical Instrumentation Group The fluorometer was equipped with a temperature control and a magnetic stirrer under the cuvette holder Wavelengths were set at 340 nm (10 nm band width) for excitation and 510 nm (20 nm band width) for emission All experiments were performed at 37 °C with constant stirring For compound studies, fura 2 loaded cells were centπfuged and resuspended in buffer A containing 1 mM CaCl2 minus BSA at 10^ cells / mL For assessment of agonist activity, a 2 mL aliquot of RBL-2H3-C3a cells was added to a cuvette and warmed in a water bath to 37 °C The 1 cm^ cuvette was transferred to the fluorometer and fluorescence was recorded for 15 seconds to ensure a stable baseline before addition of compound Fluorescence was recorded continuously for up to 2 mins after addition of compounds to monitor for the presence of any agonist activity. For antagonist studies, varying concentrations of compounds or vehicle were added to the fura 2 loaded RBL-2H3-C3a cells and monitored for 1 min to ensure that there was no change in baseline fluorescence followed by the addition of 1 nM C3a . The maximal [Ca- + ] / fura 2 fluorescence was then determined for each sample. The [Ca2 + ]j was calculated using the following formula: F - Fmin [Ca 2+ ]i = 224 (nM) -- Fmax - F The percent of maximal C3a (1 nM) induced [Ca2+]j was determined for each concentration of compound and the IC50 defined as the concentration of test compound that inhibits 50% of the maximal C3a response. Concentration response curves (5 - 7 concentrations) were run. High-Throughput-Screening-Calcium Assay: The calcium assay described above was converted to a high-throughput-screen (HTS) with the use of a 96 well Fluorescent Imaging Plate Reader (FLIPR) from Biomolecular Devices. This technology allows the measurement of the intracellular calcium mobilization in cells attached to the bottom of a 96 well plate. For this procedure, cells were obtained from the T-150 flasks as described above. The cells were plated into the 96 well plate at 30,000 cells/well. With incubation in a humidified environment in a cell incubator at 37 °C for 18-24 hours, the cells attached to the bottom surface of the 96 well plate. The FLIPR works best with the visible wavelength calcium indicators, Fluo-3 and Calcium green- 1. Both of these dyes have been used successfully for the HTS assay, but Fluo-3 was generally used. Typically 4 uM Fluo-3 was loaded into the cells for 1 hr at 37 °C in cell media without fetal calf serum and with 1.5 mM sulfinpyrazone to inhibit dye release from the cells. The media is aspirated from the cells and fresh media was added for 10 min at 37 °C to allow hydrolysis of the dye and remove extracellular dye. The media was aspirated and replaced with KRH buffer (buffer A above). After 10 min at 37 °C the cells were placed in FLIPR apparatus for analysis. FLIPR has 3-96 well plates. In addition to the plate with dye loaded cells, there is a plate containing varying concentrations of compound or vehicle and the third plate has the agonist at varying concentrations to establish agonist potency or a single concentration, e.g., 1 nM of C3a for antagonist activity. For antagonist studies, FLIPR obtains a baseline fluorescence for -30 sec, then it adds the compounds to all 96 wells simultaneously and begins to monitor changes in intracellular Ca2+. After 2 min, the contents or the agonist plate is added to the cells. The maximal Ca2+ response (in optical units) for 1 nM C3a in the presence of vehicle ( 100%) or the various concentrations of compound is determined. Inhibition curves were generated essentially as described for the single cuvette Fura-2 assay described above. The following examples are illustrative of the present invention but not intended to be limiting in any way. Example 1 ( 1 -Naphthylamino)acetylarginine a) Fmocarginine(Pbf) Wang To a mixture of Fmocarginine(Pbf) (12.2 g) and Wang resin (15 g, loading = 1.1 mmol/g) in 200 mL of CH 2 C1 2 was added EDC (4.4 g) and DMAP (400 mg). The mixture was agitated for 16 h and the liquid phase was drained. The resulting resin was washed with N-methylpyrollidine (2X) and CH2CI2 (3X). b) Bromoacetylarginine(Pbf) Wang To Fmocarginine(Pbf) Wang (5 g) was added 20% piperidine in CH2CI2 (65 mL). The mixture was agitated for 1 h and the liquid phase was drained. The resulting resin was washed with CH 2 C1 2 (5X). The resin was swelled in DMF and to the mixture was added bromoacetic acid (5.8 g) and EDC (9.1 g). The solution was agitated for 4 h and the liquid phase was drained. The resulting resin was washed with N-methylpyrollidine (2X) and CH2CI2 (4X). c) (l-Naphthylamino)acetylarginine(Pbf) Wang To bromoacetylarginine(Pbf) Wang (100 mg) in 2 mL of DMSO was added 1-naphthylamine (1 10 mg) and N-methylmorpholine (88 uL), and the mixture was heated to 50° for 16 h. The liquid phase was drained and the resin was washed with N-methylpyrollidine (2X) and CH 2 C1 2 (3X). d) ( 1 -Naphthylamino)acetylarginine (l-Naphthylamino)acetylarginine(Pbf) Wang was agitated in 5 mL of 2.5% triisopropylsilane (TIS) in 1 : 1 CH2CI2 TFA for 2 h. The liquid phase was collected in a flask and the solvent was evaporated under reduced pressure. The residue was dissolved in 1 mL of TFA and added to 7 mL of ether to precipitate out the product as a white solid. The heterogeneous mixture was centrifuged and the solvent was decanted. Ether was added to the resulting solid, the mixture centrifuged and the solvent decanted again The product was dried under vacuum ES(+) MS m e = 358 (M+H) Example 2 (3,5-Dιchlorobenzylamιno)acetylargιnιne a) (3,5-Dιchlorobenzylamιno)acetylargιnιne(Pbf)Wang Prepared according to the procedure of Example lc) except substituting 3,5-dιchlorobenzylamιne for 1-naphthylamιne and carrying out the reaction at rt b) (3.5-Dιchlorobenzylamιno)acetylargιnιne Prepared according to the procedure of Example Id) except substituting (3,5-dιchlorobenzylamιno)acetylargιnιne(Pbf) Wang for (l-naphthylamιno)acetylargιnιne(Pbf)Wang ES(+) MS m/e = 391 (M+H) Example 3 r3,5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)lacetylargιnιne la) [3,5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)1acetylargιnιne(Pbf) Wang To the title compound of Example 2a) ( 100 mg) in CH2CI2 (5 mL) was added 3-phenylpropιonyl chloride (135 uL) and N-methylmorphohne (200 uL), and the mixture was agitated for 4 h The liquid phase was drained and the resin was washed with CH 2 C1 2 (6X) b) r3.5-Dιchlorobenzylamιno-N-(3-phenylpropιonyl)lacetylargιnιne Prepared according to the procedure of Example Id) except substituting [3,5-dιchlorobenzylarnιno-N-(3-phenylpropιonyl)]acetylargιnιne(Pbf) Wang for (l-Naphthylamιno)acetylargιnιne(Pbf) Wang ES(+) MS m/e = 523 (M+H) Example 4 π,5-Dιchlorobenzylamιno-N-phenylsulfonvHacetylargιmne a) r3,5-Dιchlorobenzylamιno-N-phenylsulfonyllacetylargιnιne(Pbf) Wang To the title compound of Example 2a) ( 100 mg) in pyπdine (5 mL) was added phenylsulfonyl chloride (124 uL), and the mixture was agitated for 4 h The liquid phase was drained and the resin was washed with CH2CI2 (6X) b) r3,5-Dιchlorobenzylamιno-N-( phenylsulfonvDlacetylarginine Prepared according to the procedure of Example Id) except substituting [3,5-Dιchlorobenzylamιno-N-phenylsulfonyl]acetylargιnιne(Pbf) Wang for ( 1 -Naphthylamιno)acetylargιnιne(Pbf) Wang ES(+) MS m/e = 531 (M+H) Example 5 2-1(3.5-Dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]propιonylargιnιne a) Fmocargιnιne(Boc)τ Wang To Fmocarginine(Boc) 2 (1.8 g, 3 mmol) and Wang resin (2 g, 2 mmol) in CH2CI2 (40 mL) was added EDC (573 mg, 3mmol) and DMAP (244 mg, 2mmol). The mixture was shaken overnight and washed with DMF (2X)/CH 2 C1 (6X). b) 2-[(3,5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)lpropionic acid 3,5-dichlorobenzylamine (650 mg) and methyl 2-bromopropionate (202 uL) in THF (6 mL) was heated at reflux for 24 h under Ar. The solvent was removed under reduced pressure and residue was dried under high vacuum. The residue was dissolved in pyridine (10 mL) and treated with 3,5-dichlorophenylsulfonyl chloride (976 uL). The reaction mixture was stirred at RT overnight. Ether (50 mL) was added to the mixture, and the aqueous layer was acidified with 3N HCl. The organic layer was separated, washed with brine, and dried (MgSC>4). Silica gel flash chromatography yielded the product sulfonamide (480 mg). J H NMR (CDCI3) 5 7.2-7.8 (m, 6H), 4.71 (q, J=8.1 Hz, 1 H), 4.58 (2, J=18.9 Hz, 1H), 4.37 (d, J=18.9 Hz, 1H), 3.63 (s, 3H), 1.38 (d, J=8.1 Hz, 3H). To methyl 2-[(3,5-dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)]propionate (480 mg) in 3: 1 : 1 THF:methanol: water ( 10 mL) was added 2.5 N sodium hydroxide (1.05 mL). The solution was stirred for 1 h, CH2CI2 (50 mL) was added, and the aqueous layer acidified with 3N HCl. The organic layer was separated and dried (MgS04). c) 2-r(3,5-Dichlorobenzylamino-N-(3.5-dichlorophenylsulfonyl)1propionylarginine (Boc) Wang Fmocarginine(Boc)^Wang (500 mg) was treated with 20% piperidine in CH2CI2 (5 mL) for 30 min. The solvent was drained and the resin was washed with CH2CI2 (6X). To the resin in DMF (3 mL) was added 2-[(3,5-dichlorobenzylamino-N-(3,5- dichlorophenylsulfonyl)]propionic acid (200 mg), EDC ( 172 mg), and HOBT (122 mg), and the mixture was shaken overnight. The solution was drained, and the resin was washed with DMF (2X)/CH 2 C1 2 (6X). d) 2-r(3.5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)lpropionylarginine The resin was treated with a solution of 2.5% TIS in 1 : 1 TFA/ for 90 min. The cleavage solution was collected, the solvent removed under reduced pressure, and traces of TFA were removed by azeotroping with toluene. The residue was washed with hexanes (2X) to yield the title compound. ES(+) MS m/e = 614.1 (M+H). Example 6 [(+/-)-N-(3,5-dichlorobenzyl)prolyl]arginine a) (+/-)-Methyl N-(3,5-dichlorobenzyl)prolinate To methyl prolinate HCl (0.5 g). K2C03 (1.25 g) and 3,5-dichlorobenzylchloride (0.59 g) in DMF (5 mL) was added tetrabutylammonium iodide (cat). The solution was stirred overnight, water was added, and the mixture was extracted with ethyl acetate The organic layer was dned (Na2S04), solvent was removed under reduced pressure, and silica gel flash chromatography (0.5% methanol/CH2C12) yielded the desired product (0 65 g) ES(+) MS m/e = 288 3 (M+H) b) (+/-)-N-(3,5-dιchlorobenzyl)prolιne The title compound was prepared according to the procedure of Example 5b) except substituting (+/-)-methyl N-(3,5-dιchlorobenzyl)prolιnate for methyl 2-[(3,5- dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]propιonate ES(+) MS m e = 274 1 (M+H). c) [(+/-)-N-(3,5-Dιchlorobenzyl)prolyl]argιnιne(Boc)2 Wang The title compound was prepared according to the procedure of Example 5c) except substituting (+/-)-N-(3,5-dιchlorobenzyl)prohne for 2-[(3,5-dιchlorobenzylamιno-N-(3,5- dιchlorophenylsulfonyl)]propιonιc acid d) [(+/-)-N-(3,5-Dιchlorobenzyl)prolyl]argιnιne The title compound was prepared according to the procedure of Example 5d) except substituting [(+/-)-N-(3,5-dιchlorobenzyl)prolyl]argιnιne(Boc)2 Wang for 2-[(3,5- dιchlorobenzylamιno-N-(3,5-dιchlorophenylsulfonyl)]propιonylargιnιne (Boc) Wang The final product was puπfied by reverse phase preperative HPLC to yield the title compound. ES(+) MS m/e = 430.2 (M+H) Example 7 r(+/-)-N-(3,5-Dιchlorobenzyl)pιpecolyllacetylargιnιne The title compound was prepared according to the procedure of Example 6) except substituting methyl picohnate HCl for methyl prohnate HCl ES(+) MS m/e = 444 2 (M+H) Example 8 [3,5-Dιchlorobenzylamιno-N-(3.5-dιchlorophenylsulfonyl)lacetyl-Nα-methylargιnιne a) Nα-methylargιnιne(Mtr)OMe TFA salt To Boc-Nα-methylargιnιne(Mtr) (1 g, 2 mmol) in dry THF (50 mL) at 0°C is added diazomethane (5 mmol) in ether (10 mL) The solution was stirred for 2 h, and quenched with acetic acid. Ethyl acetate (50 mL) was added, the solution was washed with 10% sodium hydroxide and brine, and the organic layer was dried (MgSU4) Boc-Nα-methylargιnιne(Mtr)OMe was dissolved in 1.1 TFA/ at 0°C, and stirred, with warming to RT, for 2 h When the reaction appeared complete as judged by TLC, the solvent was removed under reduced pressure Trace TFA was removed by azeotroping with toluene ES(+) MS m/e = 415 4 (M+H) b) r3,5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)lacetyl-N - methylarginine(Mtr)OMe To Nα-methylarginine(Mtr)OMe TFA salt (0.48g, 0.91mmol) in 4.5 mL of DMF was added DIEA (0.24 g, 1.8 mmol). Then a premixed solution of [3,5- dichlorobenzylarnino-N-(3,5-dichlorophenylsulfonyl)]acetic acid (0.429g, 1.0 mmol), PyBOP (0.95 g, 1.8 mmol) and DIEA (0.47 g, 3.6 mmol) dissolved in 2 mL of DMF was added. The solution was stirred at RT overnight. Ethyl acetate was added, the solution was washed with 3N HCl, brine, NaHC0 3 (sat'd), and brine again. The organic layer was dried (Na2S0 4 ), filtered, and the solvent was removed under reduced pressure. The title compound was purified by silica gel chromatography using 60% ethyl acetate/hexanes to yield a light yellow foam (280 mg). ES(+) MS m e = 839.9 (M+H). c) [3,5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)1acetyl-Nα- methylarginine(Mtr) To [3,5-dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)]acetyl-Nα- methylarginine(Mtr)OMe (0.27 g, 0.323 mmol) was added 5 mL of 3: 1 : 1 THF:methanol: water. To the solution was added 2.5 N sodium hydroxide (0.26 mL, 0.646 mmol) The solution was stirred at RT for .5 h, ethyl acetate was added, and the aqueous layer acidified with 3N HCl. The organic layer was dried (Na2S0 4 ), filtered, and the solvent was removed under reduced pressure to yield a light yellow foam (0.26 mg). ES(+) MS m e = 826.0 (M+H). d) [3,5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)lacetyl-Nα- methylarginine To [3,5-Dichlorobenzylamino-N-(3,5-dichlorophenylsulfonyl)]acetyl-Nα- ethylarginιne(Mtr) (0.060 g) was added 1.5 mL of a 2.5% TIPS in 1 : 1 CH Cl2/TFA solution. The solution was stirred at RT for 4 h. The solvent was removed under reduced pressure, the title compound dissolved in a minimum of 1 : 1 TFA/CH2CI2, and a white solid was precipitated out of solution using diethyl ether. ES(+) MS m/e = 614.1 (M+H). Example 9 [N-(n-Butyl)-3-iodobenzylaminolacetylarginine a) (n-Butylamino)acetylarginine(Pbf) Wang The title compound was prepared according to the procedure of Example lc) except substituting n-butylamine for 3,5-dichlorobenzylamine. b) N-(n-Butyl)-3-iodobenzylaminolacetylarginine(Pbf) Wang To (n-Butylamino)acetylarginine(Pbf) Wang (130 mg) in DMF (3.5 mL) was added 3-iodobenzyl bromide (450 mg, 15 eq) and DIEA (0.24 mL, 15 eq). The mixture was shaken overnight, and washed repeatedly with CH2CI2. c) N-(n-Butyl)-3-iodobenzylaminolacetylarginine The title compound was prepared according to the procedure of Example Id) except substituting [N-(n-Butyl)-3-iodobenzylamino]acetylarginine(Pbf) Wang for (1- naphthylamino)acetylarginine(Pbf) Wang. The final product was purified with reverse phase preperative HPLC to yield the title compound. ES(+) MS m/e = 504.2 (M+H). Example 10 Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients. Examples of such formulations are given below. Inhalant Formulation A compound of Formula (I), (1 mg to 100 mg) is aerosolized from a metered dose inhaler to deliver the desired amount of drug per use. Example 11 Tablet Formulation Tablets/Ingredients Per Tablet 1. Active ingredient 40 mg (Cpd of Form. (I) 2. Corn Starch 20 mg 3. Alginic acid 20 mg 4. Sodium Alginate 20 mg 5. Mg stearate 1.3 mg Procedure for tablet formulation: Ingredients 1, 2, 3 and 4 are blended in a suitable mixer/blender. Sufficient water is added portion-wise to the blend with careful mixing after each addition until the mass is of a consistency to permit its conversion to wet granules. The wet mass is converted to granules by passing it through an oscillating granulator using a No. 8 mesh (2.38 mm) screen. The wet granules are then dried in an oven at 140°F (60°C) until dry. The dry granules are lubricated with ingredient No. 5, and the lubricated granules are compressed on a suitable tablet press. Example 12 Parenteral Formulation A pharmaceutical composition for parenteral administration is prepared by dissolving an appropriate amount of a compound of formula I in polyethylene glycol with heating This solution is then diluted with water for injections (to 100 mL) The solution is then rendered steπle by filtration through a 0 22 micron membrane filter and sealed in sterile containers 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 as though fully set forth

Claims: What is claimed is: 1. A compound according to Formula (I): Formula (I) wherein: A represents C j _4 alkylene, optionally substituted by C ] _4 alkyl or aryl; or A forms a 5-8 membered fused aliphatic ring with the adjacent phenyl ring; m is an integer from 1 to 3; each Rj is independently selected form the group consisting of halo, C j .4 alkyl, methanesulfonyl, alkoxy, nitrile, dimethylamine, methylenedioxy and CF3; R2 is selected from the group consisting of optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted phenyl, optionally substituted alkylsulfonyl, heteroarylsufonyl, optionally substituted phenylalkylaminocarbonyl, phenylaminocarbonyl, optionally substituted phenylalkyl, optionally substituted phenylsulfonyl, optionally substituted phenylalkylsulfonyl, optionally substituted naphthylalkylsulfonyl, heteroaryl carbonyl, heteroarylalkanoyl, optionally substituted alkylcarbonyl, optionally substituted phenylcarbonyl, and optionally substituted phenylalkylcarbonyl; wherein any substituents are selected from the group consisting of acyl, amide, C j .4 alkyl, halo, methylenedioxy, phenoxy and alkoxy; or R2 forms a 5-8 membered ring with A; or R2 forms a 4 to 7 membered ring with the carbon atom having the R4 substituent; R3 represents hydrogen or methyl; and R4 represents hydrogen or methyl. 2. A compound according to claim 1 wherein the asterisk * represents an S configuration. 3. A compound according to claim 2 wherein R3 is hydrogen. 4. A compound according to claim 3 wherein A is methylene. 5. A compound according to claim 4 wherein R j is chloro. 6. A compound according to claim 5 wherein m is 2. 7. A compound according to claim 6 wherein the Ri substituents are at the 3 and 5 positions on the aryl ring. 8. A compound according to claim 7 wherein R 4 is methyl. 9. A compound according to claim 1 selected from the group consisting of: (3,5-Dichlorobenzylamino)acetylarginine, ( 1 ,2-Diphenylethylamino)acety larginine, [ 1 -(2,2-Diphenylethylamino)]acetylarginine, [ 1 -( 1 , 2, 3,4-Tetrahydroisoquinolyl)]acety larginine, [l-Decahydroquinolinyl]acetylarginine, [ 1 -( 1 ,2,3,4-Tetrahydronaphthylamino)]acetylarginine, 3,4-Dimethylbenzylaminoacetylarginine, 3,4-Dichlorobenzylaminoacetylarginine, 3-Chlorobenzylaminoacetylarginine, 2,3-Dimethoxybenzylaminoacetylarginine, [l-(3,3-Diphenylpropylamino)]aminoacetylarginine, [l-(2-(3,4-Dimethoxyphenyl))ethylamino]aminoacetylarginine, [ 1 -(3-Phenylpropylamino)]aminoacetylarginine, Dibenzylaminoaminoacetylarginine, [3,4-Dichlorobenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, [3,4-(Methylenedioxy)benzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, 3,5-Dichlorobenzylaminoacetylarginine, [3,5-Dichlorobenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-phenoxypropionyl)]acetylarginine, [3,4-Dichlorobenzylamino-N-(2-(4-chlorophenoxy)acetyl)]acetylarginine, and [3,5-Dichlorobenzylamino-N-(3-phenylpropionyl)]acetylarginine. 10. A compound according to claim 9 selected from the group consisting of: [3,5-Dichlorobenzylamino-N-(3-pyridinecarbonyl)]acetylarginine, [3,5-Dichlorobenzylamino-N-(2-phenoxypropionyl)]acetylarginine, (3,5-Dichlorobenzylamino)acetylarginine, and [3,5-Dichlorobenzylamino-N-(3-phenylpropionyl)]acetylarginine. 11. [3,5-Dichlorobenzylamino-N-(3-phenylpropionyl)]acetylarginine. 12. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier. 13. A method of antagonizing a C3A receptor which comprises administering to a subject in need thereof, an effective amount of a compound of claim 1. 14. A method of treating an immune or inflammatory disease or disorder characterized by abnormal levels of anaphylatoxins which comprises administering to a subject in need thereof an effective amount of a compound of claim 1. 15. A method according to claim 14 wherein the disease or disorder is selected from the group consisting of rheumatoid arthritis, Alzheimer's disease, psoriasis, gout, multiple sclerosis, systemic lupus erythematosus, glomerulonephritis and adult respiratory distress syndrome.

Inventors: LEE DENNIS, LEE, DENNIS
Applicants: LEE DENNIS, SMITHKLINE BEECHAM CORP, LEE, DENNIS, SMITHKLINE BEECHAM CORPORATION
Application year: 1998
Main classification: C07C53
IPC codes: A61K31, A61P11, A61P13, A61P17, A61P19, A61P25, A61P29, A61P37, A61P43, C07C279, C07C311, C07D211, C07D213, C07D215, C07D217, C07D233, C07D317, C07D333, C07D405