EP1150955A2 - Composes de fixation au recepteur heptahelicoidal couple a la proteine g et leurs procedes d'utilisation - Google Patents

Composes de fixation au recepteur heptahelicoidal couple a la proteine g et leurs procedes d'utilisation

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Publication number
EP1150955A2
EP1150955A2 EP00907184A EP00907184A EP1150955A2 EP 1150955 A2 EP1150955 A2 EP 1150955A2 EP 00907184 A EP00907184 A EP 00907184A EP 00907184 A EP00907184 A EP 00907184A EP 1150955 A2 EP1150955 A2 EP 1150955A2
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Prior art keywords
compound
protein coupled
moiety
coupled heptahelical
disorder
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German (de)
English (en)
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Kazumi Shiosaki
Paul Fleming
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Millennium Pharmaceuticals Inc
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Millennium Pharmaceuticals Inc
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/57—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of rings other than six-membered aromatic rings
    • C07C233/60—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of rings other than six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/64—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings
    • C07C233/67—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
    • C07C233/75—Carboxylic acid amides having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/49—Carboxylic acid nitriles having cyano groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C255/57—Carboxylic acid nitriles having cyano groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing cyano groups and carboxyl groups, other than cyano groups, bound to the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/62—Oxygen or sulfur atoms
    • C07D213/63—One oxygen atom
    • C07D213/65—One oxygen atom attached in position 3 or 5
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
    • C07D213/82—Amides; Imides in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated

Definitions

  • chemokine family of peptides is defined on the basis of sequence homology and on the presence of variations on a conserved cysteine motif (Schall (1996) Cytokine 3: 165-183; and Oppenheim et al. ( ⁇ 99 ) Annu. Rev. Immunol 9:617-648).
  • the family can be subdivided on the basis of this motif into two major subfamilies, in which members of each contain four characteristic cysteine residues. This subdivision therefore defines the CC or ⁇ -chemokine family in which the first two cvsteines are juxtaposed, and the CXC or ⁇ -chemokine family in which there is an intervening amino acid between the first two cysteines.
  • Chemokines display a range of in vitro and in vivo functions ranging from proinflammatory activities on a range of cell types to proliferative regulatory activities. All of the functions of the chemokine family are believed to be signaled into a responsive cell using members of the G protein-coupled heptahelical receptor family. To date several ⁇ -chemokine and ⁇ -chemokine receptors have been described, (e.g.. Neote et al. (1993) Cell 72:415-425; Ponath et al. (1996) J. Exp. Med. 183:2437-2448; and Power et al. (1995) J Biol. Chem. 270:19495-19500.)
  • the present invention is based, at least in part, on the discovery of compounds which interact with G-protein coupled heptahelical receptors.
  • the compounds of the invention can be used to treat chemokine mediated disorders, e.g. neurological, immunological, inflammatory and cancer related disorders.
  • the present invention provides a G-protein coupled heptahelical receptor (GPCR) binding compound of the formula: J-M (I) where J is an aromatic moiety and M is a G-protein coupled heptahelical receptor pocket interacting moiety.
  • GPCR G-protein coupled heptahelical receptor
  • the compound interacts with a ⁇ -chemokine receptor, e.g. CCR2, CCR3, CCR4, CCR5. CCR6, CCR7. CCR8, or CCR10.
  • the compound modulates recruitment of at least one cell type. e.g. leukocytes, e.g. macrophages or eosinophils. associated with inflammation in a subject.
  • the invention in another aspect, pertains to a method for treating a chemokine mediated disorder in a subject.
  • the method involves administering an effective amount of a G-protein coupled heptahelical receptor binding compound such that the disorder is treated, e.g. at least one symptom of the disorder is diminished or alleviated.
  • the chemokine mediated disorder may be a neurological disorder (e.g. multiple sclerosis, Alzheimer's disease, or Parkinson's disease), an immunological disorder (e.g. AIDS, arthritis, or lupus), cancer, or an inflammatory disorder (e.g. asthma).
  • the invention pertains to a compound represented by the formula:
  • A is selected from the group consisting of branched and straight chain alkyl, aryl, alkenyl, alkynyl, and heteroaryl moieties optionally substituted by NR'R", CN, NO2, F, Cl, Br, I, CF3, CCI3, CHF 2 .
  • Li is a linker moiety selected from the group consisting of a bond, O, S, CHOH, CHSH, CHNH 2 , CHNHR, CHNRR * , NH, NR, (CH 2 ) n , O(CH 2 ) n , and (CH 2 ) n O(CH 2 ) n , an optionally substituted ring moiety of 4 to 7 atoms containing up to three heteroatoms, a chain of 1 to 5 atoms optionally substituted by C ⁇ -Cg alkyl, halogens, wherein n is either 0, 1, 2, or 3, and R and R' are each independently substituted or unsubstituted C1 - C branched or straight chain alkyl, C1 -Cg branched or straight chain alkenyl, aryl, C4- C7 ring, optionally substituted with up to three heteroatoms; - j -
  • B is an aromatic moiety containing from 0 to 3 heteroatoms and containing 5 to 7 members optionally substituted by NR'R", cyano, nitro. halogen, CF3, CHF 2 , CONR'R", S(O)NR'R", CHO, OCF3, SCF3, COR', CO 2 R, OR where R and R" are each independently hydrogen, halogen, C1 -Cg alkyl, optionally substituted aryl or optionally substituted aryl;
  • E is a G-protein coupled heptahelical receptor pocket interacting moiety.
  • the invention also relates to a compound represented by the formula:
  • Z ⁇ , Z 2 , Z3, and Z4 are each independently N or C; R ⁇ , R 2 , R3, R4, R5, Rg, R7, and Rg are each independently hydrogen,
  • Li is O, S, NH, NR7, (CH 2 ) n , CO, CHOH, O(CH ) n , and (CH 2 ) n O(CH 2 ) n wherein n is either 1,2, or 3 and R7 is C ⁇ -Cg branched or straight chain alkyl, alkoxy, thioalkyl, hydroxyalkyl, halo, haloalkyl, amino, alkylamino, or carboxyl; and
  • the invention relates to a pharmaceutical preparation comprised of the G-protein coupled heptahelical receptor binding compound and a pharmaceutically acceptable carrier.
  • the invention also pertains to a packaged G- protein coupled heptahelical receptor binding compound containing instructions for using the compound for treating a chemokine mediated disorder.
  • Figure 1 depicts a binding curve of Compound A in a DB Assay (see Example 3 below).
  • Figure 2 depicts a binding curve of Compound CU in a DB Assay (see Example 3 below).
  • Figure 3 depicts a binding curve of Compound CV in a DB Assay (see Example 3 below).
  • Figure 4 is a graph depicting the blockage of THP.l cell migration by compound B in a CBIR Assay (see Example 4 below).
  • Figure 5 is a graph depicting the blockage of THP.l cell migration by compound
  • Figure 6 shows MCP-5 induced peritoneal eosinophil recruitment in mice after administering compounds B and C in a MIR Assay (see Example 5 below).
  • the present invention pertains to a G-protein coupled heptahelical receptor binding compound of the formula:
  • G protein coupled heptahelical receptor includes receptors for which belong to the G-protein coupled receptor (GPCR) superfamily of seven- transmembrane domain heptahelical receptors.
  • GPCRs G-protein coupled receptors
  • GPCRs G-protein coupled receptors
  • effectors intracellular enzymes and channels which are modulated by G-proteins
  • the GPCR protein superfamily now contains over 250 types of paralogues, receptors that represent variants generated by gene duplications (or other processes), as opposed to orthologues, the same receptor from different species.
  • the superfamily can be broken down into five families: Family I, receptors typified by rhodopsin and the beta2-adrenergic receptor and currently represented by over 200 unique members (reviewed by Dohlman et al. (1991) Annu. Rev. Biochem. 60:653-688 and references therein); Family II, the recently characterized parathyroid hormone/calcitonin/secretin receptor family (Juppner et al. (1991) Science 254:1024-1026; Lin et al.
  • GPCRs include chemokine receptors which are expressed in specific tissues and leukocyte subtypes. Many chemokine receptors can bind to and be activated by more than one chemokine, and many chemokines can bind and activate more than one receptor in the nanomolar or subnanomolar range (MacKay (1996) J. Ex Med. 184:522-549; Wells et al. (1996) Chem. Biol. 3:603-609). Chemokine receptors can be classified generally into three groups: ⁇ -chemokine receptors, ⁇ - chemokine receptors and ⁇ - ⁇ chemokine receptors.
  • the GPCR binding compounds of the present invention interact with receptors of the ⁇ -chemokine receptor family (Bonini et al. DNA Cell Biol. (1997) 16(10): 1249- 1256).
  • ⁇ -chemokine receptors include CCR2. CCR3, CCR4. CCR5, CCR6. CCR7, CCR8, and CCR10.
  • the ⁇ -chemokine receptors (CCRs) are characterized by their ability to bind to CC chemokines (also referred to as ⁇ -chemokines).
  • the CC chemokines are characterized by a conserved cysteine motif, in which the first two cysteines are juxtaposed.
  • the CCR family includes both specific and non-specific receptors.
  • CCR1 is known to bind macrophage inflammation protein- la (MlP-la),
  • RANTES regulation on activation normal T cell expressed and secreted
  • MCP-3 monocyte chemoattractant protein-3
  • CCR2 binds MCP-1 , MCP-3, and MCP-4 (Myers et al. (1995) J. Biol. Chem. 270:5786-5792; Garcia- Zepeda et al. (1996) J. Immunol. 157:5613), whereas CCR3 recognizes eotaxin and MCP-4 (Kitaura et al. (1996) J. Biol. Chem. 271 :7725; Garcia-Zepada et al. (1996) J. Immunol.
  • CCR4 is activated by macrophage inflammatory protein-la (MIP- la), RANTES, and MCP-1 (Power et al. (1995) J. Biol. Chem. 270:19495).
  • CCR5 was found to bind to and be activated by RANTES , MlP-la, and MlP-lb (Raport et al. (1996) J. Biol. Chem. 271 : 17161).
  • CCR6 and CCR7 have recently been discovered and specifically bind to liver and activation regulated chemokine (L ARC) and EBI 1 -ligand chemokine (ELC) respectively (Baba et al, 1997; Yoshida et al. 1997). It is known that CCR10 binds MCP-1 and MCP-3 with high affinity.
  • aromatic moiety includes groups with aromaticity, e.g. moieties that have at least one aromatic ring.
  • aromaticity e.g. moieties that have at least one aromatic ring.
  • 5- and 6-membered single-ring aromatic groups that may include from zero to four heteroatoms.
  • Aryl groups also include polycyclic fused aromatic groups such as naphthyl, quinolyl, indolyl, and the like.
  • aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles", “heteroaryls” or “heteroaromatics”.
  • the aromatic ring can be substituted at one or more ring positions with such substituents as described above, as for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato.
  • phosphinato cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino. arylcarbonylamino, carbamoyl and ureido), amidino, imino. sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl, sulfonamido, nitro. trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
  • Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings which are not aromatic so as to form a polycycle (e.g., tetralin).
  • the aromatic moiety of the present invention comprises two aromatic rings, e.g. pyridyl or pyrimidyl rings, e.g. two pyrimidyl rings connected with an ether linkage.
  • G-protein coupled heptahelical receptor pocket refers to a region of the GPCR which is capable of interacting with, e.g. binding to. the GPCR pocket binding moiety. Not wishing to be bound by theory, it is believed that the GPCR pocket may be a cavity in the GPCR lined with hydrophobic amino acid residues.
  • G-protein coupled heptahelical receptor pocket interacting moiety refers to a group which interacts with a GPCR pocket.
  • the group may be substituted or unsubstituted aromatic, alkyl, alkenyl, cycloalkyl, etc.
  • the interaction between the GPCR pocket interacting moiety and the pocket includes any interaction which allows the compound to perform its intended function, e.g., the interaction is hydrophobic, ionic, covalent, or combinations thereof.
  • the GPCR binding compound modulates the recruitment of at least one inflammatory cell type in a subject.
  • the ability of a GPCR binding compound to modulate the recruitment of at least one inflammatory cell type can be measured or observed using art-recognized techniques or assays. Examples of such assays are the Murine Inflammatory Cell Recruitment Assay (referred to herein as the MICR Assay) and the Cell Based Inflammatory Recruitment Assay (referred to herein as the CIBR Assay) as described in Examples 4 and 5, respectively.
  • the term "subject” includes any animal which expresses GPCRs, for example, mammals e.g., mice, rats, cows, sheep, pigs, horses, monkeys, dogs, cats and, preferably, humans.
  • the language "inflammatory cell type” includes cell types associated with a chemokine mediated disorder characterized by inflammation. Examples of these cell types include, but are not limited to, leukocytes, e.g., eosinophils, neutrophils, basophils, fibroblasts, monocytes, T lymphocytes, and macrophages.
  • leukocytes e.g., eosinophils, neutrophils, basophils, fibroblasts, monocytes, T lymphocytes, and macrophages.
  • Leukocytes are white blood cells which are involved in nonspecific resistance against pathogenic microorganisms and inflammatory response. Monocytes are particularly important in the nonspecific immune response, while lymphocytes are especially important in the specific immune response.
  • Neutrophils are the most abundant phagocytic cells in blood and are continuously produced in circulating blood, affording protection against the entry of foreign materials.
  • leukocytes exhibit chemotaxis and are attracted to foreign substances, including invading microorganisms, which they engulf and digest along with particulate matter.
  • Eosinophils are leukocytes which react with the acidic dye eosin.
  • Basophils are leukocytes which stain with basic dyes.
  • Macrophages are large ameboid mononuclear phagocytic cells.
  • the present invention also pertains to a GPCR binding compound which is an antagonist of a G-protein coupled heptahelical receptor, e.g., a ⁇ -chemokine receptor.
  • the IC 50 of the GPCR binding compound to the GPCR is about 10 ⁇ M or less, e.g., about 5 ⁇ M or less, e.g., about 1 ⁇ M or less, e.g., about 50 nM or less.
  • the term "antagonist" includes compounds which bind to the GPCR such that the binding of a second compound to the GPCR is modulated.
  • the ability of a compound to bind to a GPCR can be determined through using art-recognized techniques and assays.
  • assays include the Time Resolved Fluorescence assay (herein referred to as the TRF assay) and the Direct Binding Assay (herein referred to as the DB Assay), described in Examples 2 and 3, respectively.
  • the TRF assay determines the binding affinity of a compound to a receptor by over expressing the receptor in a culture of cells.
  • the TRF assay determines the binding affinity of a compound to a receptor using a cell line engineered to overexpress a GPCR, e.g. CCR10. The cells are exposed simultaneously to the test compound and a fluorescently labeled ligand specific for the receptor.
  • the invention pertains to a method for treating a chemokine mediated disorder, (e.g., a neurological disorder, an immunological disorder, a disorder characterized by inflammation, or a disorder characterized by unwanted cellular proliferation) in a subject.
  • a chemokine mediated disorder e.g., a neurological disorder, an immunological disorder, a disorder characterized by inflammation, or a disorder characterized by unwanted cellular proliferation
  • the method includes administering an effective amount of a G-protein coupled heptahelical receptor binding compound to a subject.
  • the disorder may be treated through modulation of a ⁇ -chemokine receptor, e.g., CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, or CCR10.
  • a ⁇ -chemokine receptor e.g., CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, or CCR10.
  • preferred disorders include AIDS, multiple sclerosis, asthma, cancer, and lupus.
  • the disorder may also be characterized by abnormal cellular signal transduction, or amounts of chemokine stimulated chemotaxis.
  • administering includes routes of administration which allow the GPCR binding compound to perform its intended function, e.g. interacting with GPCRs and/or treating a chemokine mediated disorder.
  • routes of administration which can be used include parental injection (e.g., subcutaneous, intravenous, and intramuscular), intraperitoneal injection, oral, inhalation, and transdermal.
  • the injection can be bolus injections or can be continuous infusion.
  • the GPCR binding compound can be coated with or disposed in a selected material to protect it from natural conditions which may detrimentally effect its ability to perform its intended function.
  • the GPCR binding compound can be administered alone or with a pharmaceutically acceptable carrier.
  • the GPCR binding compound can be administered as a mixture of GPCR binding compounds, which also can be coadministered with a pharmaceutically acceptable carrier.
  • the GPCR binding compound can be administered prior to the onset of a chemokine mediated disorder, or after the onset of a chemokine mediated disorder.
  • the GPCR binding compound also can be administered as a prodrug which is converted to another form in vivo.
  • treatment includes the diminishment or alleviation of at least one symptom associated or caused by the disorder being treated.
  • treatment can be diminishment of several symptoms of a disorder or complete eradication of a disorder.
  • chemokine mediated disorder includes a disorder characterized by the participation of chemokines or association with chemokines.
  • the language also includes disorders characterized by aberrant chemokine expression.
  • Chemokines have a wide variety of functions. They are able to elicit chemotactic migration of distinct cell types, such as monocytes, neutrophils, T lymphocytes, basophils and fibroblasts. Many chemokines have proinflammatory activity and are involved in multiple steps during an inflammatory reaction.
  • chemokines have been shown to exhibit other activities.
  • macrophage inflammatory protein -1 MIP-1
  • platelet factor-4 is a potent inhibitor of endolethial cell growth
  • interleukin-8 promotes proliferation of keratinocytes
  • GRO is an autocrine growth factor for myeloma cells.
  • Chemokines have been proposed to participate in a number of physiological and disease conditions, including, for example, lymphocyte trafficking, wound healing, hemapoietic regulation and immunological disorders such as asthma and arthritis.
  • chemokine mediated disorder characterized by inflammation includes a disorder having inflammation as at least one of its symptoms.
  • disorders include anaphylaxis, systemic necrotizing vasculitis. systemic lupus erthyematosus, serum sickness syndromes, psoriasis, rheumatoid arthritis, adult respiratory distress syndrome (ARDS), allergic rhinitis, atopic dermatitis, asthma and other allergic responses, and reperfusion injury occurring after periods of ischemia such as in myocardial infarction or shock.
  • the disorder is asthma.
  • chemokine mediated disorders include neurological related disorders, immunological related disorders and disorders characterized by unwanted cellular proliferation, e.g. cancer.
  • Neurological related disorders includes disorders of the nervous system, including, but not limited to those involving the brain, the central and peripheral nervous system, and the interfaces between muscles and the nerves.
  • Some examples of neurological related disorders include Alzheimer's disease, dementias related to Alzheimer's disease (such as Pick's disease), Parkinson's and other Lewy diffuse body diseases, multiple sclerosis, amyotrophic lateral sclerosis, progressive supranuclear palsy, epilepsy, and Jakob-Creutzfieldt disease.
  • Neurological related disorders also includes neurological disorders associated with inflammation, e.g. stroke, traumatic injury to the brain, traumatic injury to the spinal cord, spinal crush, and central and peripheral nervous system trauma.
  • immunological related disorder includes both organ-specific and systemic immunological disorders.
  • immunological disorders include immune thyroiditis, hyperthyroidism, type I diabetes mellitus, insulin related diabetes, Addison's disease, autoimmune oophoritis, autoimmune orchitis, AIDS, autoimmune hemolytic anemia, paroxysmal cold hemoglobinuria, autoimmune thrombocytopenia, autoimmune neutropenia, pernicious anemia, autoimmune coagulopathies, myasthenia gravis, multiple sclerosis, experimental allergic encephalomyelitis, pemphigus and other bullous diseases, rheumatic carditis, Goodpasture's syndrome, postcardiotomy syndrome, systemic lupus erythematosus, rheumatoid arthritis, keratitis, parotitis, polymositis, dermatomyositis, and scleroderma.
  • the immunological disorder is AIDS, multiple sclerosis
  • the invention pertains to a pharmaceutical preparation comprised of an effective amount of a G-protein coupled heptahelical receptor binding compound and a pharmaceutically acceptable carrier.
  • the effective amount is an effective amount to treat a ⁇ -chemokine mediated disorder, e.g., asthma.
  • pharmaceutically acceptable carrier includes substances capable of being coadministered with the GPCR binding compound(s), and which allows both to perform their intended function, e.g., treating a chemokine mediated disorder or preventing a chemokine mediated disorder.
  • examples of such carriers include solutions, solvents, dispersion media, delay agents, emulsions and the like. The use of such media for pharmaceutically active substances are well known in the art. Any other conventional carrier suitable for use with the GPCR binding compound also fall within the scope of the present invention.
  • pharmaceutically acceptable carrier is art recognized and includes a pharmaceutically acceptable material, composition or vehicle, suitable for administering compounds of the present invention to mammals.
  • the carriers include liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject agent from one organ, or portion of the body, to another organ, or portion of the body.
  • Each carrier must be "acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • materials which can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth: malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'
  • wetting agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.
  • antioxidants examples include: water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT). lecithin, propyl gallate, ⁇ -tocopherol, and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.
  • Formulations of the present invention include those suitable for oral, nasal, topical, transdermal.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred per cent, this amount will range from about 1 per cent to about ninety-nine percent of active ingredient, preferably from about 5 per cent to about 70 per cent, most preferably from about 10 per cent to about 30 per cent.
  • Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
  • Formulations of the invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient.
  • lozenges using a flavored basis, usually sucrose and acacia or tragacanth
  • a compound of the present invention may also be administered as a bolus, electuary or paste.
  • solid dosage forms of the invention for oral administration capsules, tablets, pills, dragees, powders, granules and the like
  • the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol.
  • binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; absorbents, such as kaolin and bentonite clay; lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and coloring agents.
  • binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/
  • compositions may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
  • a tablet may be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent.
  • Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • the tablets, and other solid dosage forms of the pharmaceutical compositions of the present invention may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres.
  • compositions may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use.
  • These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner.
  • embedding compositions which can be used include polymeric substances and waxes.
  • the active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.
  • Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluent commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluent commonly used in the art, such as, for example, water or other solvents, solubilizing agents and e
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar- agar and tragacanth, and mixtures thereof.
  • Formulations of the pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the invention with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
  • suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
  • Formulations of the present invention which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.
  • Dosage forms for the topical or transdermal administration of a compound of this invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.
  • the active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.
  • the ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients, such as animal and vegetable fats. oils, waxes. paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones. bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • excipients such as animal and vegetable fats. oils, waxes. paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones. bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
  • Powders and sprays can contain, in addition to a compound of this invention.
  • excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.
  • Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
  • Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body.
  • dosage forms can be made by dissolving or dispersing the compound in the proper medium.
  • Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the active compound in a polymer matrix or gel.
  • compositions of this invention suitable for parenteral administration comprise one or more compounds of the invention in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use.
  • sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use.
  • antioxidants antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
  • aqueous and nonaqueous carriers examples include water, ethanol. polyols
  • glycerol such as glycerol, propylene glycol, polyethylene glycol. and the like
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
  • adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.
  • Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride
  • the absorption of the drug in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
  • Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissue. The preparations of the present invention may be given orally, parenterally, topically, or rectally. They are of course given by forms suitable for each administration route.
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular. subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • systemic administration means the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient's system and. thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • These compounds may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intracistemally and topically, as by powders, ointments or drops, including buccally and sublingually.
  • the compounds of the present invention which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art.
  • Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex. weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.
  • a physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.
  • the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical composition.
  • certain embodiments of the present compounds can contain a basic functional group, such as amino or alkylamino, and are, thus, capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids.
  • pharmaceutically acceptable salts is art recognized and includes relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed.
  • Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J Pharm. Sci. 66:1-19).
  • the compounds of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases.
  • pharmaceutically acceptable salts in these instances includes relatively non-toxic, inorganic and organic base addition salts of compounds of the present invention. These salts can likewise be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine.
  • Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like.
  • Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine. ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.
  • esters refers to the relatively non-toxic, esterified products of the compounds of the present invention. These esters can be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in its free acid form or hydroxyl with a suitable esterifying agent.
  • Carboxylic acids can be converted into esters via treatment with an alcohol in the presence of a catalyst.
  • Hydroxyls can be converted into esters via treatment with an esterifying agent such as alkanoyl halides.
  • the term also includes lower hydrocarbon groups capable of being solvated under physiological conditions. e.g., alkyl esters, methyl, ethyl and propyl esters. (See, for example, Berge et al., supra.)
  • a preferred ester group is an acetomethoxy ester group.
  • the language "effective amount" of the compound is that amount necessary or sufficient to treat or prevent a chemokine mediated disorder, e.g. prevent the various mo ⁇ hological and somatic symptoms of a chemokine mediated disorder.
  • the effective amount can vary depending on such factors as the size and weight of the subject, the type of illness, or the particular GPCR binding compound. For example, the choice of the GPCR binding compound can affect what constitutes an "effective amount".
  • One of ordinary skill in the art would be able to study the aforementioned factors and make the determination regarding the effective amount of the GPCR binding compound without undue experimentation.
  • the regimen of administration can affect what constitutes an effective amount.
  • the GPCR binding compound can be administered to the subject either prior to or after the onset of a chemokine mediated disorder. Further, several divided dosages, as well as staggered dosages, can be administered daily or sequentially, or the dose can be continuously infused, or can be a bolus injection. Further, the dosages of the GPCR binding compound(s) can be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.
  • the invention features a protein coupled heptahelical receptor binding compound comprising a G-protein coupled heptahelical receptor binding compound packaged with instructions for using said compound for treating a ⁇ - chemokine mediated disorder.
  • the invention also features a method of using a G-protein coupled heptahelical receptor binding compound to modulate the binding of a second compound to a G- protein coupled heptahelical receptor.
  • the invention pertains to a compound represented by the formula:
  • A-L ⁇ -B-L 2 -E (II) where A is selected from the group consisting of branched and straight chain alkyl, aryl, alkenyl, alkynyl, and heteroaryl moieties optionally substituted by NR'R", CN, NO 2 , F, Cl, Br, I, CF 3 , CC1 3 , CHF 2 , CHCl 2 , CONR'R", S(O)NR'R", CHO, OCF3, OCCI3, SCF3, SCCI3, COR', CO 2 R, and OR' and wherein R and R" are each independently hydrogen, C ⁇ -Cg alkyl, C -C£ alkenyl, or optionally substituted aryl;
  • Li is a linker moiety selected from the group consisting of a bond, O, S, CHOH, CHSH, CHNH 2 , CHNHR, CHNRR', NH, NR, (CH 2 ) n , O(CH 2 ) n , and (CH 2 ) n O(CH 2 ) n , an optionally substituted ring moiety of 4 to 7 atoms containing up to three heteroatoms, a chain of 1 to 5 atoms optionally substituted by C1 -C alkyl.
  • halogens wherein n is either 0, 1, 2, or 3, and R and R' are each independently substituted or unsubstituted C1 - C ⁇ branched or straight chain alkyl, C ⁇ -C branched or straight chain alkenyl, aryl, C4- C7 ring, optionally substituted with up to three heteroatoms; B is an aromatic moiety containing from 0 to 3 heteroatoms and containing 5 to 7 members optionally substituted by NR'R", cyano. nitro. halogen. CF3. CHF 2 . CONR'R", S(O)NR'R". CHO. OCF3, SCF3. COR. CO 2 R, OR where R and R" are each independently hydrogen, halogen. C1-C6 alkyl, optionally substituted aryl or optionally substituted aryl;
  • L 2 is a second linking moiety selected from the group consisting of a bond. CH9CO. NHCO, OCO, CO, CH 2 NHCO, CHOH. (CH 2 ) n . O. NH. 0(CH ? ) n . NH(CH 2 ) n . CH 2 CHOH and NRCO; and
  • E is a G-protein coupled heptahelical receptor pocket interacting moiety.
  • L is S, NH, CH 2 , or O.
  • L 2 is NHCO.
  • A is represented by the following formula:
  • Z, and Z 2 each independently represent N or C;
  • R,, R 2 , and R 3 are independently selected from the group comprised of hydrogen, C,-C 12 branched or straight chain alkyl. alkoxy, thioalkyl, hydroxyalkyl, halo, haloalkyl, amino, alkylamino. or carboxyl.
  • both Z, and Z 2 are carbon
  • R is alkyl (e.g.. methyl), halogen (e.g., bromine, chlorine or fluorine), or alkoxy and the L, linker is located in the meta position.
  • R is alkyl (e.g.. methyl), halogen (e.g., bromine, chlorine or fluorine), or alkoxy and the L, linker is located in the meta position.
  • R is alkyl (e.g.. methyl), halogen (e.g., bromine, chlorine or fluorine), or alkoxy and the L, linker is located in the meta position.
  • halogen e.g., bromine, chlorine or fluorine
  • R is carbonyl (e.g., a ketone. an aldehyde, an ester, or an amide.).
  • R may be substituted with a cyclic moiety such as piperazine, furan, or phenyl.
  • A is substituted or unsubstituted phenyl. Examples of substituents includes substituted or unsubstituted alkyl (e.g.. methyl), alkenyl, aryl and heteroaryl moieties.
  • A may be substituted with halogens (e.g., chlorine).
  • L is O.
  • the invention features a compound wherein B is represented by the following formula:
  • Z 3 and Z 4 each independently represent N or C
  • R 4 and R 5 are independently selected from the group comprised of hydrogen, C,-C 6 branched or straight chain alkyl, alkenyl, alkynyl. alkoxy. thioalkyl, hydroxyalkyl, halo, haloalkyl, amino, alkylamino, or carboxyl.
  • R 4 is alkyl and R 5 is hydrogen.
  • B is a substituted or unsubstituted pyridyl or pyrimidyl moiety, wherein B may be represented by the following formula:
  • the invention features a compound wherein E is represented by the formula below:
  • R 6 is an electron withdrawing moiety and the aryl ring is additionally optionally substituted with zero to four halogen atoms.
  • E is substituted with at least one fluorine atom, e.g. two or more fluorine atoms.
  • R 6 may be alkyl, alkoxy, haloalkyl, nitro. halo, alkylamino. hydroxyalkyl. or carboxyl.
  • E is a para substituted aryl moiety represented by the formula below:
  • R 6 is a halogenated alkyl moiety, e.g. a fluorinated alkyl moiety, e.g. a trifluoromethyl or pentafluoroethyl.
  • R 5 include substituted or unsubstituted alkoxy moieties (e.g.. methoxy. trifiuoromethoxy) or thioether moieties.
  • R 6 can be alkenyl or alkynyl (e.g.. ethenyl).
  • E is heterocyclic, e.g.. substituted or unsubstituted furanyl, imidazolyl. benzothiophenyl. benzylfuranyl, quinolinyl, isoquinolinyl, benzodioxazolyl, benzoxazolyl, benzothiazolyl, benzylimidazolyl, thiazolyl, isothiaozolyl, oxazolyl, benzylthiazolyl, isooxazolyl, methylenedioxyphenyl, indolyl, thienyl, pyrimidyl, pyrazinyl, purinyl, or deazapurinyl.
  • benzothiophenyl benzylfuranyl, quinolinyl, isoquinolinyl, benzodioxazolyl, benzoxazolyl, benzothiazolyl, benzylimidazolyl, thiazolyl, iso
  • E is branched or straight chain alkenyl or alkynyl.
  • examples include ethynyl trimethyl silane and alkenes (e.g., dienes, trienes).
  • the invention features a compound represented by the formula below:
  • Z,, Z 2 , Z 3 , and Z 4 are each independently N or C;
  • R,, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , and R 8 are each independently hydrogen, C,-C 6 branched or straight chain alkyl, alkenyl, alkynyl. alkoxy, thioalkyl, hydroxyalkyl, halo, haloalkyl, amino, alkylamino, or carboxyl;
  • L is O, S, NH, NR 7 , (CHR 7 ) n , CO. CR 7 OH, O(CHR 7 ) n , and (CHR 7 ) n O(CHR 7 ) n wherein n is either 1,2, or 3;
  • L 2 is a second linking moiety selected from the group consisting of a bond, CH 2 CO, NHCO, OCO. CO, CH 2 NHCO. NHCOCH 2 . CHOH, (CH 2 ) n , O. NH, O(CH 2 ) m , NH(CH 2 ) n ⁇ , CH 2 CHOH and NRCO. wherein m is 0, 1, 2, or 3.
  • the compound may be represented by the formula below:
  • Z1 , Z 2 . Z3, and Z4 are each independently N or C; R ⁇ , R 2 , R3, R4, R5, Rg, R7, and Rg are each independently hydrogen,
  • Li is O, S, NH, NR 7 , (CH 2 ) n , CO, CHOH, O(CH 2 ) n , and (CH 2 ) n O(CH 2 ) n wherein n is either 1,2, or 3 and R7 is C1 -Cg branched or straight chain alkyl, alkoxy, thioalkyl, hydroxyalkyl, halo, haloalkyl, amino, alkylamino, or carboxyl;
  • L 2 is a second linking moiety selected from the group consisting of a bond.
  • Z, and Z 2 are both carbon.
  • R, is methyl, and R 2 and R 3 are hydrogen.
  • L, is O.
  • R 4 and R 5 are both hydrogen.
  • L 2 is NHCO.
  • R 6 is a halogenated alkyl moiety, e.g. a fluorinated alkyl moiety, e.g. a trifluoromethyl or pentafluoroethyl moiety.
  • R 7 and R s are each independently fluorine or hydrogen.
  • the invention can be represented by the structure below:
  • the invention also features compounds of the formula:
  • L is O and L 2 is NHCO.
  • Li is O
  • L is NHCO
  • Rg is a halogen, a halogenated alkyl group, e.g. trifluoromethyl or pentafluoroethyl), or an alkoxy group, e.g. a halogenated alkoxy group, e.g. a trifluoromethoxy group.
  • R 6 also can be ethenyl or a thioether moiety (e.g. -S-CF 3 ).
  • the invention also features binding compounds represented by the following structures:
  • alkyl includes saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.
  • alkyl further includes alkyl groups, which can further include oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more carbons of the hydrocarbon backbone, e.g., oxygen, nitrogen, sulfur or phosphorous atoms.
  • a straight chain or branched chain alkyl has 10 or fewer carbon atoms in its backbone (e.g., C ] - C ⁇ o for straight chain, C3-C1 Q for branched chain), and more preferably 6 or fewer.
  • preferred cycloalkyls have from 4-7 carbon atoms in their ring structure, and more preferably have 5 or 6 carbons in the ring structure.
  • alkyl includes both "unsubstituted alkyls" and “substituted alkyls”, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone.
  • substituents can include, for example, halogen, hydroxyl, alkylcarbonyloxy.
  • arylcarbonyloxy alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino. arylcarbonylamino, carbamoyl and ureido), amidino, imino.
  • sulfhydryl alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, silyl, trialkylsilyl, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
  • moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.
  • Cycloalkyls can be further substituted, e.g., with the substituents described above.
  • An "alkylaryl" moiety is an alkyl substituted with an aryl (e.g., phenylmethyl (benzyl)).
  • aryl includes aryl groups, including 5- and 6-membered single-ring aromatic groups that may include from zero to four heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, benzoxazole, benzothiazole, triazole, tetrazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
  • Aryl groups also include polycyclic fused aromatic groups such as naphthyl, quinolyl, indolyl, and the like.
  • aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles", “heteroaryls” or “heteroaromatics".
  • the aromatic ring can be substituted at one or more ring positions with such substituents as described above, as for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy. arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl.
  • alkylthiocarbonyl phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido).
  • Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings which are not aromatic so as to form a polycycle (e.g... tetralin
  • alkenyl and alkynyl include unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
  • lower alkyl as used herein means an alkyl group, as defined above, but having from one to three carbon atoms in its backbone structure. Likewise, “lower alkenyl” and “lower alkynyl” have similar chain lengths.
  • alkoxy alkyl include alkyl groups, as described above, which further include oxygen, nitrogen or sulfur atoms replacing one or more carbons of the hydrocarbon backbone, e.g.. oxygen, nitrogen or sulfur atoms.
  • polycyclyl or “polycyclic radical” refer to two or more cyclic rings
  • cycloalkyls e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls
  • rings e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls
  • bridged rings Rings that are joined through non-adjacent atoms are termed "bridged" rings.
  • Each of the rings of the polycycle can be substituted with such substituents as described above.
  • halogen hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato.
  • cyano amino (including alkyl amino, dialkylamino, arylamino, diarylamino. and alkylarylamino), acylamino (including alkylcarbonylamino. arylcarbonylamino.
  • heteroatom as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus. It will be noted that the structure of some of the compounds of this invention includes asymmetric carbon atoms.
  • the present invention includes GPCR binding compounds and/or methods of using the same which are encompassed by the formulae set forth herein and which are not described in Brombridge. et al., J. Med Chem. (1997) 40:3494, U.S. 3,499,898, EP358 1 18, EP 344 634, and/or WO 96/23783. The contents of each of which are expressly inco ⁇ orated herein by reference.
  • E or the G-protein coupled heptahelical pocket interacting moiety is not:
  • L is O
  • L 2 is NHCO
  • E or the G-protein coupled heptahelical pocket interacting moiety is not:
  • E or the G-protein coupled heptahelical pocket interacting moiety is a phenyl group having at least one substituent.
  • E is a phenyl group having at least one substituent in the para position.
  • E is a phenyl group having at least two substituents.
  • the compounds of the present invention can be synthesized using standard methods of chemical synthesis and/or can be synthesized using schemes described herein. Synthesis of specific compounds is discussed in detail in the Example sections below. An example of a general synthesis is outlined in the scheme below: 52 -
  • Hydroxypyridine (or other hydroxyaryl precursor) is dissolved in dioxane and 1.5 equivalents of 95% sodium hydride is added. The mixture is stirred at room temperature for 20 minutes and 1 equivalent of 2-chloro-5-nitropyridine (or other ⁇ - chloro-heterocycle) is then added. The mixture is subsequently brought to reflux for 3 hours and cooled. The reaction mixture is then quenched by addition of saturated ammonium chloride solution. Silica gel is added to the solution and the mixture is rotovapped to dryness. The product is eluted from the silica gel and flash chromatographed using a mixture of ethyl acetate/hexane.
  • the nitro group is reduced by dissolving the nitropyridine in 1 :1 methanol: water. Acetic acid and iron powder is then added and the mixture is brought to reflux for 3 hours. After cooling, the iron is precipitated by addition of 20% NaOH and subsequently filtered through Celite. The methanol is removed by rotary evaporation and the remaining aqueous mixture is extracted with methylene chloride. The organic layer is dried and the solvent removed by rotary evaporation to give product.
  • dipyridyl ether is then dissolved in methylene chloride followed the addition of polymer bound mo ⁇ holine (Booth, et al., J. Am. Chem. Soc, 119, 1997, 4882-4886). 1.3 equivalents of acid chloride is added and the mixture shaken overnight. The excess acid chloride is scavenged using polymer bound tris-2-aminoethylamine. Filtration followed by flash chromatography of the filtrate using ethyl acetate :hexane gives the product.
  • hydroxypyridine (or other hydroxyaryl precursor) is dissolved in 500 mL of dioxane. 1.5 equivalents of 95% sodium hydride is added and the mixture stirred at room temperature for 20 minutes. 1 equivalent of 2-chloro-5-nitropyridine (or other ⁇ - chloro-heterocycle) is added and the mixture is brought to reflux for 3 hours. After cooling, the reaction mixture is quenched by addition of 2 mL of saturated ammonium chloride solution. 10 g of silica gel is added to the solution and the mixture is rotovapped to dryness. The product is eluted from the silica gel and flash chromatographed using a mixture of ethyl acetate/hexane.
  • CCR10 cells are stable recombinant K293 cells overexpressing the CCR10 receptor.
  • the cells are routinely cultured and passaged in a growth medium composed of DMEM base medium: 10%) fetal bovine serum (FBS), IX Glutamine, and 0.4 mg/ml G418. 1% Pen/Strep is also included in the media when the cells are seeded into the plates on Day 1.
  • DMEM base medium 10%
  • FBS fetal bovine serum
  • IX Glutamine IX Glutamine
  • G418 fetal bovine serum
  • Pen/Strep is also included in the media when the cells are seeded into the plates on Day 1.
  • rhMCP-1 carrier free was supplied as a frozen stock solution in PBS at a concentration of 0.65 mg/ml. 250 ⁇ g (385 ⁇ l) was buffer exchanged on a 2.8 ml column of Sephardex G-25 (fine), equilibrated with lOmM Borax, pH 9. Fractions (250 ⁇ l) were collected and 2 ⁇ l aliquots were analyzed using a Bradford protein assay. Four fractions eluting immediately after the void volume contained the bulk of the protein and were pooled (approx. 1 ml). 1 mg of Eu-labeling reagent (1.5 ⁇ mol) was dissolved in 200 ⁇ l of water and 100 ⁇ l of this solution was added to the protein pool. The final relative concentration ratios of Eu:MCP-l was about 26:1. The ratio of Eu:NH 2 was about 3:1.
  • the mixture was incubated for 20 hours at 4°C and then desalted on a 15 mL column of Sephardex G-25 (fine) equilibrated with 20 mM HEPES (hemi-sodium salt), pH 7.5. and 0.9% NaCl.
  • 0.5 mL fractions were collected and 1 ⁇ l aliquots were added to 100 ⁇ l of enhancement solution and counted on the VICTOR.
  • the five fractions that were eluted immediately after the void volume showed a peak of Eu containing material and were pooled. 60 ⁇ l of the pool was set aside for protein assay. 36 ⁇ l of 0.1% BSA (heavy metal free; Wallac) was added as a stabilizer to the remainder.
  • the material was then stored at 4°C. The total volume was about 2.5 ml.
  • the MCP-1 concentration was determined by a BCA protein assay with BSA as a standard to be 8.5 ⁇ M (74 ⁇ g/ml).
  • the Eu concentration of the material was measured on the VICTOR with a standard curve prepared from the Eu standard solution provided from the vendor, Wallac.
  • the Eu concentration was 12 ⁇ M.
  • the calculated stoichiometry was 1.4 Eu chelate per MCP-1 molecule.
  • the compounds were provided as a dried film on a polystyrene "Master Plate” containing 1 ⁇ g of compound per well.
  • the compounds were dispensed into cell plates by preparing a 3X stock solution in binding buffer from the dried films.
  • the cell plates were prepared according to the following method. Prior to the day of dispensation of compounds onto cells, the "Master Plates" were allowed to reach room temperature. 50 ⁇ l each of IX Binding Buffer (0.125% BSA in deionized water) and MCP-1 stock solution (40nM and 600 nM in IX Binding Buffer) was added to each well. The plates were subsequently stored at 4° C overnight.
  • TRF Time-resolved fluorescence
  • Each 96-well tissue culture plate (poly-D-lysine coated) was seeded with 40.000 cells in complete media containing 0.4 mg/ml G418 (200 ⁇ l per well). The plates were incubated at 37° C and 6% CO 2 and left overnight. Day 2
  • the cells were approximately 90%> confluent in the wells by Day 2.
  • the cells were washed twice with Binding Buffer (no BSA) using 200 ⁇ l per wash. 40 ⁇ l of Binding Buffer was left behind following the second wash. To this final 40 ⁇ l. 100 ⁇ l of Binding Buffer was then added. Excess Binding Buffer from each well was removed leaving about 15 ⁇ l in each well. 15 ⁇ l of compound solution was dispensed into each well followed by 15 ⁇ l of 30 nM Eu-MCPl working solution. The plates were then incubated at room temperature for 3 hours. The plates were then washed twice with 150 ⁇ l of Wash Buffer (250 mM Hepes.
  • Controls were included on each 96- well assay plate.
  • the controls were (1 ) wells with no compounds and no unlabeled MCPl, (2) wells with no compounds and unlabeled MCPl (13.3 nM) corresponding to the IC 50 , and (3) wells with no compounds and unlabeled MCPl (200 nM) corresponding to complete inhibition. Each control was set up in duplicates.
  • TRF Time-resolved fluorescence
  • CCR10 receptors were expressed in stably transfected K293 cells, and the cells were maintained in DMEM base medium, 10% FBS, IX glutamine, and 0.4 mg/ml G418, and in the incubator set at 37 °C, 6.0 % CO? and 90% relative humidity. The day before the experiment, the cells were trypsinized and 200 ⁇ l of the cell suspension (150,000 cells/ml) was deposited into 96-well Biocoat plates (poly-D-lysine-coated). The binding assay was performed 24 hours later.
  • 8-point dose response curves were generated as follows: compounds to be tested were dissolved in DMSO at 10 mg/ml concentration and diluted to 100 ⁇ g/ml into n- butanol. 0.75, 1.5, 3, 6, 12, 18, 24, and 30 ⁇ l of the compounds were dispensed into 8 wells of a 96-well Costar plate. In the case of more potent compounds, further dilution was applied to generate 10 ⁇ g/ml compound solutions in n-butanol, and the solutions were used to make the compound plates by dispensing 1, 2, 4, 8, 10, 20 ⁇ l of the diluted solutions, and 4 and 8 ⁇ l of the 100 ⁇ g/ml solutions, into 8 wells of the plate.
  • the compound plates were placed in the hood overnight to evaporate the butanol, leaving dried films of the compounds.
  • 50 ⁇ l of filtered binding buffer (25 mM HEPES, pH 7.4, 75 ⁇ M EDTA, 1 1.5 mM KC1, 115 mM NaCl, 6 mM MgSO4, and 1.8 mM CaCl 2 ) was added into each well, and the plates were stored at 4 °C for about two hours.
  • the compounds were thoroughly resuspended in the binding buffer. The cells were washed three times with the binding buffer by adding and then decanting the medium or binding buffer and drying the plates on paper towels.
  • the buffer was added slowly to the side wall of the wells to avoid disturbing the cells.
  • the plates were dried over paper towels to leave about 5 ⁇ l of the binding buffer in each well.
  • An additional 10 ⁇ l of binding buffer was added to each well.
  • 15 ⁇ l of the compound solutions, and then 15 ⁇ l of the Eu ⁇ + -i aDe ⁇ ec ! MCP-1 ligand solution (15 nM) in binding buffer with 0.1% BSA was added.
  • the binding reaction mixtures were maintained at RT for three hours.
  • a THP-1 cell line was used that expresses both CCR-10 and CCR-2.
  • the cells were placed in the top half of a chamber (Transwell plate, 24- well, 5 ⁇ M pore size, purchased from Costar) separated in the middle by a membrane.
  • a gradient of chemokine MCP-1 or MCP-3, purchased from R&D was established which, in the absence of inhibitors, leads to migration of cells across the membrane. Quantitation of cell migration was done using a FACScan machine. The % migration was calculated as the number of migrated cells divided by the number of input cells. As shown in Figures 4 and 5, compounds B and C blocked chemokine induced migration of cells.
  • MIR Assay Murine Inflammatory Recruitment Assay
  • CCR-10 and its ligands MCP-1, MCP-3 (MCP-5 in mice) have been demonstrated to mediate the recruitment of eosinophils and a variety of other leukocytes to tissue where it is expressed.
  • Antibodies against CCR-10 have been shown to block the effects of these ligands.
  • This example demonstrates that compounds B and C are capable of blocking MCP-5 induced peritoneal eosinophil recruitment.
  • mice 8-10-wk-old C57BL/6J mice were purchased from the Jackson laboratory (Bar Harbor, ME) and kept in Millennium Pharmaceuticals Inc. Specific Pathogen Free mouse facility.
  • T-lymphocytes, B-lymphocytes and mononuclear phagocytes were identified by Thy 1.2 (53-2.1) (PharMingen, San Diego, CA), IgM (11/41) (PharMingen, San Diego, CA) and Moma-2 (Biosource Int. Camarillo, CA) staining.
  • Percentage of eosinophils, lymphocytes, neutrophils and macrophages was determined by counting their number in eight high power fields (40x magnification; total area 0.5 mm2) per area randomly selected and dividing this number by the total number of cells per high power field. To obtain the absolute number of each leukocyte subtype in the lavage, these percentages were multiplied by the total number of cells recovered from the peritoneal fluid. As shown in Figure 6, mice pretreated with compounds B and C prior to MCP-5 challenge showed significantly reduced levels of eosinophil recruitment than untreated mice. Control experiments with eotaxin demonstrate that the compounds are acting through CCR-10 rather than by inhibition of cytoskeletal function.

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Abstract

L'invention concerne des composés de fixation au récepteur heptahélicoïdal couplé à la protéine G, notamment, des composés représentés par la formule: J-M dans laquelle J représente une fraction aromatique et M représente une fraction interactive avec une poche du récepteur heptahélicoïdal couplé à la protéine G. On peut utiliser ces composés afin de traiter des maladies provoquées par l'intermédiaire de chimiokines, telles que des maladies neurologiques, immunologiques, inflammatoires et apparentées au cancer.
EP00907184A 1999-02-04 2000-02-03 Composes de fixation au recepteur heptahelicoidal couple a la proteine g et leurs procedes d'utilisation Withdrawn EP1150955A2 (fr)

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Families Citing this family (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6316503B1 (en) 1999-03-15 2001-11-13 Tularik Inc. LXR modulators
WO2002064136A2 (fr) 2001-01-26 2002-08-22 Chugai Seiyaku Kabushiki Kaisha Inhibiteurs de malonyl-coa decarboxylase utiles en tant que modulateurs metaboliques
ATE359773T1 (de) 2001-01-26 2007-05-15 Chugai Pharmaceutical Co Ltd Verfahren zur behandlung von erkrankungen mit malonyl coa-decarboxylase-inhibitoren
CN101596191B (zh) 2001-01-26 2011-06-15 中外制药株式会社 用作代谢调节剂的丙二酰基辅酶a脱羧酶抑制剂
EP1379243B1 (fr) 2001-02-20 2009-09-09 Chugai Seiyaku Kabushiki Kaisha Methodes de traitement de maladies metaboliques dans lesquelles sont utilisees des inhibiteurs de malonyl-coa decarboxylase
US7709510B2 (en) 2001-02-20 2010-05-04 Chugai Seiyaku Kabushiki Kaisha Azoles as malonyl-CoA decarboxylase inhibitors useful as metabolic modulators
JP4434745B2 (ja) 2002-01-30 2010-03-17 アムジェン インコーポレイテッド 複素環式アリールスルホンアミドベンジル化合物
ES2314212T3 (es) 2002-01-30 2009-03-16 Amgen Inc. Compuestos arilsulfonamidobencilicos.
US7205307B2 (en) 2002-02-14 2007-04-17 Icagen, Inc. Pyrimidines as novel openers of potassium ion channels
CA2533749C (fr) 2003-08-01 2012-07-10 Chugai Seiyaku Kabushiki Kaisha Composes de piperidine utiles en tant qu'inhibiteurs de malonyl-coa decarboxylase
US7285562B2 (en) 2003-08-01 2007-10-23 Chugai Seiyaku Kabushiki Kaisha Cyanoamide compounds useful as malonyl-CoA decarboxylase inhibitors
CA2533747C (fr) 2003-08-01 2012-11-13 Chugai Seiyaku Kabushiki Kaisha Composes heterocycliques utiles comme inhibiteurs de la malonyl-coa decarboxylase
ATE407673T1 (de) 2003-08-01 2008-09-15 Chugai Pharmaceutical Co Ltd Azol-verbindungen auf cyanoguanidin-basis als malonyl-coa decarboxylase-hemmer
US7037927B2 (en) * 2003-10-16 2006-05-02 Abbott Laboratories Amides that inhibit vanilloid receptor subtype 1 (VR1) receptor
EP1773797A2 (fr) 2004-08-06 2007-04-18 Otsuka Pharmaceutical Co., Ltd. Compose aromatique
ATE462697T1 (de) * 2004-08-12 2010-04-15 Amgen Inc Bisaryl-sulfonamide
NZ556017A (en) * 2004-12-24 2009-10-30 Prosidion Ltd G-protein coupled receptor (gpr116) agonists and use thereof for treating obesity and diabetes
CN101355939A (zh) * 2005-10-06 2009-01-28 马萨诸塞大学 用于抑制hiv复制的新物质的组合物和合成
ES2463450T3 (es) 2005-12-05 2014-05-28 Otsuka Pharmaceutical Co., Ltd. Fármaco medicinal
US8030487B2 (en) * 2006-07-07 2011-10-04 Targegen, Inc. 2-amino—5-substituted pyrimidine inhibitors
UA95978C2 (ru) 2006-10-02 2011-09-26 Оцука Фармас'Ютікел Ко., Лтд. Ингибитор активации stat3/5
UA102514C2 (ru) * 2007-02-22 2013-07-25 Сингента Партисипейшнс Аг Производные иминопиридина, промежуточное соединение, способ борьбы с заражением полезных растений грибами или его предупреждение и композиция на основе производной иминопиридина
JP2009185020A (ja) * 2008-01-11 2009-08-20 Ishihara Sangyo Kaisha Ltd ピリジルアミジン誘導体又はその塩、並びにそれらを有効成分として含有する農園芸用殺菌剤
ES2645689T3 (es) 2008-05-21 2017-12-07 Ariad Pharmaceuticals, Inc. Derivados de fósforo como inhibidores de quinasas
US9273077B2 (en) 2008-05-21 2016-03-01 Ariad Pharmaceuticals, Inc. Phosphorus derivatives as kinase inhibitors
US11351168B1 (en) 2008-06-27 2022-06-07 Celgene Car Llc 2,4-disubstituted pyrimidines useful as kinase inhibitors
US8338439B2 (en) 2008-06-27 2012-12-25 Celgene Avilomics Research, Inc. 2,4-disubstituted pyrimidines useful as kinase inhibitors
NZ589843A (en) 2008-06-27 2012-12-21 Avila Therapeutics Inc Pyrimidine heteroaryl compounds and uses thereof as protein kinase inhibitors
KR101705158B1 (ko) 2009-05-05 2017-02-09 다나-파버 캔서 인스티튜트 인크. Ｅｇｆｒ 억제제 및 질환 치료방법
US8962656B2 (en) * 2010-06-01 2015-02-24 Boehringer Ingelheim International Gmbh CCR2 antagonists
DK2603081T3 (en) 2010-08-10 2017-01-16 Celgene Avilomics Res Inc COLLECTED BY A BTK INHIBITOR
TWI545115B (zh) 2010-11-01 2016-08-11 阿維拉製藥公司 雜環化合物及其用途
JP5956999B2 (ja) 2010-11-01 2016-07-27 セルジーン アヴィロミクス リサーチ， インコーポレイテッド ヘテロアリール化合物およびその使用
CA2816957A1 (fr) 2010-11-07 2012-05-10 Targegen, Inc. Compositions et procedes de traitement de la myelofibrose
US8796255B2 (en) 2010-11-10 2014-08-05 Celgene Avilomics Research, Inc Mutant-selective EGFR inhibitors and uses thereof
GB201104153D0 (en) 2011-03-11 2011-04-27 Glaxo Group Ltd Novel compounds
EA201391626A1 (ru) 2011-05-04 2014-03-31 Ариад Фармасьютикалз, Инк. Соединения для ингибирования клеточной пролиферации в egfr-стимулированных типах рака
EP2770830A4 (fr) 2011-10-28 2015-05-27 Celgene Avilomics Res Inc Méthodes de traitement d'une maladie ou d'une affection associée à la tyrosine-kinase btk (bruton's tyrosine kinase)
ES2698298T3 (es) 2012-03-15 2019-02-04 Celgene Car Llc Sales de un inhibidor de quinasa receptor de factor de crecimiento epidérmico
EP2825041B1 (fr) 2012-03-15 2021-04-21 Celgene CAR LLC Formes solides d'un inhibiteur de kinases du récepteur du facteur de croissance épidermique
AU2013204563B2 (en) 2012-05-05 2016-05-19 Takeda Pharmaceutical Company Limited Compounds for inhibiting cell proliferation in EGFR-driven cancers
ITMI20120785A1 (it) * 2012-05-09 2013-11-10 Fond Italiana Sclerosi M Ultipla Fism Onlu Modulatori del recettore gpr17
EP2935226A4 (fr) 2012-12-21 2016-11-02 Celgene Avilomics Res Inc Composés hétéroarylés et leurs utilisations
CA2900012A1 (fr) 2013-02-08 2014-08-14 Celgene Avilomics Research, Inc. Inhibiteurs d'erk et leurs utilisations
MY191087A (en) 2013-03-15 2022-05-30 Global Blood Therapeutics Inc Compounds and uses thereof for the modulation of hemoglobin
US9611283B1 (en) 2013-04-10 2017-04-04 Ariad Pharmaceuticals, Inc. Methods for inhibiting cell proliferation in ALK-driven cancers
US9492471B2 (en) 2013-08-27 2016-11-15 Celgene Avilomics Research, Inc. Methods of treating a disease or disorder associated with Bruton'S Tyrosine Kinase
EA201992707A1 (ru) 2013-11-18 2020-06-30 Глобал Блад Терапьютикс, Инк. Соединения и их применения для модуляции гемоглобина
US9415049B2 (en) 2013-12-20 2016-08-16 Celgene Avilomics Research, Inc. Heteroaryl compounds and uses thereof
EP3179858B1 (fr) 2014-08-13 2019-05-15 Celgene Car Llc Formes et compositions d'un inhibiteur d'erk
GB201516504D0 (en) 2015-09-17 2015-11-04 Astrazeneca Ab Imadazo(4,5-c)quinolin-2-one Compounds and their use in treating cancer
KR20180094923A (ko) 2015-12-24 2018-08-24 교와 핫꼬 기린 가부시키가이샤 α, β 불포화 아미드 화합물
CN114149341B (zh) * 2021-12-08 2023-07-21 中国药科大学 芳氧环己基酰胺类ampk激动剂及其制备方法和医药用途

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3804346A1 (de) * 1988-02-12 1989-08-24 Boehringer Mannheim Gmbh Tert.-butylphenyl-pyridyl-amide - verfahren zu ihrer herstellung sowie diese verbindungen enthaltende arzneimittel
GB9025123D0 (en) * 1990-11-19 1991-01-02 Ici Plc Nitrogen compounds
EP0863757B1 (fr) * 1995-08-10 2003-03-26 Merck & Co., Inc. Pyrroles d'aryle substitues en position 2, compositions contenant de tels composes et leurs modes d'utilisation
US6022884A (en) * 1997-11-07 2000-02-08 Amgen Inc. Substituted pyridine compounds and methods of use
AR015733A1 (es) * 1998-03-25 2001-05-16 Otsuka Pharma Co Ltd DERIVADO DE PIRIDINA, EL PRODUCTO Y LA COMPOSICIoN FARMACEUTICA QUE CONTIENE DICHO DERIVADO.
ES2265923T3 (es) * 1999-03-24 2007-03-01 Anormed Inc. Compuestos heterociclicos que se unen a receptores de quimiocinas.

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0046203A2 *

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