Method of identifying inhibitors of C-C chemokine receptor 3
AssigneesChildren's Medical Center Corporation, The Brigham and Women's Hospital, Inc., Millennium Pharmaceuticals, Inc.
InventorsCraig J. Gerard, Norma P. Gerard, Charles R. Mackay, Paul D. Ponath, Theodore W. Post, Shixin Qin
Life of this patent
Estimating the term…
Abstract
The present invention relates to a method of identifying an inhibitor of a mammalian (e.g., human) C-C chemokine receptor 3 (CCR3), comprising combining (a) a compound to be tested, (b) a host cell expressing a recombinant protein comprising a mammalian CCR3, and (c) a ligand of said receptor, under conditions suitable for binding of ligand to said receptor, and detecting or measuring the formation of a receptor-ligand complex. Inhibition of complex formation by the compound is indicative that the compound is an inhibitor.
Claims52
An isolated nucleic acid, wherein said isolated nucleic acid hybridizes to a nucleic acid selected from the group consisting of:
- a) a nucleic acid having the sequence set forth in SEQ ID NO:
- 1 or a portion thereof comprising the coding sequence;
- b) a nucleic acid having the sequence set forth in SEQ ID NO:
- 3 or a portion thereof comprising the coding sequence;
- c) a nucleic acid having the sequence set forth in SEQ ID NO:
- 5 or a portion thereof comprising the coding sequence;
- d) the complement of any one of (a) through (c);
- and
- e) the RNA counterpart of any one of (a) through (d), wherein U is substituted for T.
An isolated, recombinant nucleic acid which encodes a mammalian chemokine receptor-3.
An isolated nucleic acid, wherein said nucleic acid encodes a polypeptide selected from the group consisting of:
- a) a polypeptide having the amino acid sequence set forth in SEQ ID NO:2;
- b) a polypeptide having the amino acid sequence set forth in SEQ ID NO:4;
- c) a polypeptide having the amino acid sequence set forth in SEQ ID NO:6;
- and
- d) a functional portion of any one of (a) through (c), said portion having at least one function characteristic of a mammalian C-C chemokine receptor.
A recombinant construct comprising a recombinant nucleic acid which hybridizes to a nucleic acid selected from the group consisting of:
- a) a nucleic acid having the sequence set forth in SEQ ID NO:
- 1 or a portion thereof comprising the coding sequence;
- b) a nucleic acid having the sequence set forth in SEQ ID NO:
- 3 or a portion thereof comprising the coding sequence;
- c) a nucleic acid having the sequence set forth in SEQ ID NO:
- 5 or a portion thereof comprising the coding sequence;
- d) the complement of any one of (a) through (c);
- and
- e) the RNA counterpart of any one of (a) through (d), wherein U is substituted for T.
A recombinant construct comprising a nucleic acid which encodes a mammalian chemokine receptor 3.
A recombinant construct comprising a nucleic acid, wherein said nucleic acid encodes a polypeptide selected from the group consisting of:
- a) a polypeptide having the amino acid sequence set forth in SEQ ID NO:2;
- b) a polypeptide having the amino acid sequence set forth in SEQ ID NO:4;
- c) a polypeptide having the amino acid sequence set forth in SEQ ID NO:6;
- d) a functional portion of any one of (a) through (c), said portion having at least one function characteristic of a mammalian C-C chemokine receptor;
- and
- e) a functional equivalent of any one of (a) through (c).
An isolated, recombinant mammalian chemokine receptor 3 or portion thereof, said portion having at least one function characteristic of a mammalian chemokine receptor or an immunological property.
An essentially pure mammalian chemokine receptor 3 or portion thereof, having at least one function characteristic of a mammalian chemokine receptor, for example, exotaxin binding.
A host cell containing an isolated, recombinant nucleic acid encoding a mammalian CKR-3 receptor protein or functional portion thereof.
A fusion protein containing a mammalian chemokine receptor 3 protein or portion thereof, said portion having at least one functional characteristic of a mammalian chemokine receptor or an immunological property.
A method for producing a mammalian chemokine receptor 3 protein or functional portion thereof comprising maintaining a host cell containing a recombinant nucleic acid encoding said receptor or functional portion thereof under conditions suitable for expression of the nucleic acid, whereby the encoded protein is expressed and said receptor or portion thereof is produced.
A method for producing a mammalian chemokine receptor 3 protein or functional portion thereof comprising the following steps:
- a) providing a recombinant nucleic acid construct comprising a nucleic acid which encodes all or part of a coding sequence for a mammalian chemokine receptor 3 protein wherein the coding sequence is operably linked to at least one expression control sequence;
- b) introducing the construct into a suitable host cells;
- and
- c) maintaining the host cells in suitable medium under conditions whereby the nucleic acid is expressed.
An antibody or functional portion thereof which binds to a mammalian chemokine receptor 3 protein or portion of said receptor protein.
A method of detecting a ligand of a mammalian chemokine receptor 3 protein or portion thereof comprising the steps of combining a compound to be tested with an active, isolated mammalian chemokine receptor 3 protein under conditions suitable for binding of ligand thereto, and detecting or measuring the formation of a complex between said compound and the active, isolated protein.
A method of identifying a ligand of a mammalian chemokine receptor 3 protein or portion thereof comprising the steps of:
- a) combining a compound to be tested with a host cell expressing an active, recombinant mammalian chemokine receptor 3 protein under conditions suitable for binding of ligand thereto;
- and
- b) detecting or measuring the formation of a complex between said compound and the active, isolated protein.
A method of identifying an inhibitor of a mammalian chemokine receptor 3 protein or portion thereof comprising the steps of:
- a) combining a compound to be tested with a host cell expressing an active, recombinant mammalian chemokine receptor 3 protein under conditions suitable for binding of ligand thereto in the presence of a ligand;
- and
- b) detecting or measuring the formation of a complex between said ligand and the active, isolated protein.
An inhibitor of at least one function characteristic of a mammalian chemokine receptor 3 protein.
Antisense nucleic acid comprising a nucleic acid having a sequence that hybridizes to a nucleic acid having a sequence selected from the group consisting of:
- a) SEQ ID NO:
- 1;
- b) SEQ ID NO:
- 3;
- c) SEQ ID NO:
- 5;
- b) an RNA counterpart of any one of a) through c).
A method of modulating at least one function of a mammalian chemokine receptor 3 protein, comprising the step of contacting said protein with an inhibitor or promoter of at least one function of said protein.
A method for treating an inflammatory disease or condition, comprising administering to a mammal a therapeutically effective amount of an inhibitor of a mammalian chemokine receptor 3 protein, whereby inflammation is reduced.
An antibody having binding specificity for a mammalian chemokine receptor 3 protein or portion thereof, wherein the antibody blocks binding of a ligand to the receptor and inhibits function associated with binding of the ligand to the receptor.
Dependencies are inferred from the claim text.